Topic 1: Experiences and challenges in implementing new techniques for the detection and identification of LMOs

a) a) Yes, there are new analytical approaches developed since approximately 2023 that are directly applicable to the detection and identification of GMOs/LMOs (organisms modified by transgenesis or by new genomic techniques – NGT).
1. New PCR / dPCR methods specifically targeting genome-edited plants (NGT)
1.1. Event- / mutation-specific qPCR for genome-edited plants (e.g., FAD2 soybean)
• Heinz et al., 2025 developed and validated two event-specific qPCR methods to detect small base-pair deletions in the FAD2-1A and FAD2-1B genes of Calyno soybean, a CRISPR-edited variety.
• Principle:
o Primers overlap the exact deletion site, preventing amplification of the wild-type allele (allele-specific PCR strategy).
o Use of LNA bases in the forward primer to increase discrimination between the edited and wild-type alleles.
o The same hydrolysis probe is used for both edited variants to reduce cost.
• Performance:
o LOD95 ≈ 3–5 copies per reaction, qPCR efficiency ≈ 95–97%, R² ≈ 0.999, interlaboratory validation following ENGL criteria.
o Reference: J Consum Prot Food Saf 20, 53–62 (2025). https://doi.org/10.1007/s00003-024-01538-0
1.2. Event-specific qPCR methods for new “classical” GM events
Since 2023, several new GM events have been the subject of validated qPCR methods, for example:
• An event-specific qPCR method for GM maize CC-2, validated through a multi-laboratory collaborative trial (Frontiers in Plant Science, 2024. https://doi.org/10.3389/fpls.2024.1460038)
• An EURL GMFF (2024) report on the quantification of maize event DP23211, with a LOD around 0.04% (in copy number), fully compliant with ENGL requirements for official control.
(Reports available at https://joint-research-centre.ec.europa.eu)
These new protocols can be incorporated into screening matrices for seeds or imported products (maize, soybean, canola).
1.3. Multiplex droplet digital PCR (ddPCR) for detecting several events in a single reaction
Since 2023, there has been significant progress in multiplex ddPCR:
• Verginelli et al., 2024: internal validation of four duplex ddPCR assays (event + reference gene) enabling quantification of multiple GM maize/soybean events with high repeatability and linearity in the 0.1–5% mass fraction range (Foods 2024, 13(24), 4011. https://doi.org/10.3390/foods13244011).
• A 2025 study demonstrated the simultaneous detection of 2–4 soybean or canola GM events using multiplex ddPCR on the QX200 system (0.1–5%), while maintaining accurate quantification (
Journal of AOAC International, Oct. 2024; 108(1). DOI:10.1093/jaoacint/qsae082 ).
ddPCR offers very robust absolute quantification, less affected by amplification efficiency than qPCR.
Multiplexing reduces both screening time and costs (several LMOs detected in a single test).
2. Targeted and untargeted NGS (HTS) approaches adapted to NGT detection
2.1. Interlaboratory reliability of NGS methods for GMOs
• Pallarz et al., 2023 demonstrated interlaboratory reproducibility of NGS methods — particularly hybridization probe capture + sequencing — to detect GM sequences in rapeseed-based animal feed, including NGT-edited rapeseed (
https://doi.org/10.1093/jaoacint/qsae082).
• ENGL (2025) integrated these findings into a report on detecting organisms produced via NGT (initially microorganisms), concluding that probe-capture + NGS is transferable and repeatable, while emphasizing the need for standardization (minimum sensitivity, validation requirements).
(Reports available at: https://joint-research-centre.ec.europa.eu)

2.2. NGT in commercial products: combined targeted PCR + NGS approaches
• Guertler et al., 2023 analysed the detection of marketed plant products obtained through NGT by combining specific qPCR assays and targeted NGS for crops such as SU Canola (AHAS gene editing).
They showed that, when full sequence information is available, robust detection methods can be developed for commercialized NGT products (
https://doi.org/10.1016/j.foodcont.2023.109869).
3. CRISPR/Cas-based biosensors and rapid tests for LMOs
This is likely the most striking innovation since 2023 for rapid GMO/LMO detection.
3.1. CRISPR/Cas12a electrochemical biosensor for MON810 maize
• Zhu et al., 2024 described an electrochemical CRISPR/Cas12a biosensor for detecting MON810 maize.
• Features:
o Combination of RPA (isothermal amplification) + Cas12a + nanomaterial-based electrochemical architecture.
o Rapid and highly sensitive detection of the transgenic Cry1Ab/Ac sequence, with strong potential for large-scale field testing.
( https://doi.org/10.1016/j.aca.2024.342290).
3.2. CRISPR/Cas12a + RPA/LAMP + lateral flow assay (LFA) for CP4-EPSPS, Cry1Ab/Ac
• A 2025 review on GMO detection perspectives reports several RPA-Cas12a GM tests, including:
o RPA + Cas12a for CP4-EPSPS and Cry1Ab/Ac, with lateral-flow readout,
o LOD ≈ 45 copies/µL, results within ~45 min, readable by the naked eye.
(https://doi.org/10.1007/s00604-025-07267-x).
• Additional studies detail LAMP–CRISPR/Cas12a–LFA assays with fluorescent visualization and strip-based detection for the same resistance genes.
These systems are particularly suitable for field pre-screening (ports, borders, fields) for classical GM traits (glyphosate tolerance, insect resistance).
3.3. PCR / LAMP assays for genome-edited plants (CRISPR)
• Singh et al., 2025 developed a detection method for a CRISPR-Cas9–edited tomato (deletion in the SlPL gene).
o Development of a deletion-specific multiplex qPCR (detection down to 0.1% of the edited line)
o Plus LAMP/PCR assays targeting the Cas9 gene to confirm the absence of residual transgene cassettes (J Genet Eng Biotechnol. 2025 Oct 29;23(4):100609. doi:10.1016/j.jgeb.2025.100609).

b) The transition to the use of new methodologies has not been completed in my laboratory.
c) NA
d) The factors that have prevented the adoption of new methodologies are the lack of technical and human capacity building. This requires substantial financial resources.

Dear Mr. David,
Greetings! Thank you so much for the opportunity to discuss my experiences in the GMO detection laboratory. I truly appreciated being part of such an important discussion.
To address your specific questions:

a) No new analytical techniques were developed. Our GMO detection analysis still continues relying on conventional PCR using a thermocycler.

b) Our laboratory has not yet transitioned to newer methodologies for several reasons. The lab was not fully established in previous years but has recently been reorganized and has now commenced GMO testing.

d) Reasons for Not Implementing New Methodologies
The implementation of advanced methodologies, such as digital and quantitative PCR (qPCR), has been delayed due to the following challenges:
   1. Lack of Access: We have been unable to acquire the necessary equipment and reagents for new methodologies like digital and qPCR.
   2. Lack of Training: We have not received training on the adoption and application of these new techniques.
   3. Delay in Establishment of Laboratory: The process of establishing a well-organized GMO detection laboratory experienced significant delays.
   4. Procurement Challenges: Our national procurement system has made it difficult to reliably obtain high-quality instruments and reagents in a timely manner.

Dear Mr. Bulubulu Otono,

Thank you for your contribution.
You have nicely summarized some of the newly developed methodologies. You've mentioned lack of resources (human, technical and financial) that prevent you from implementation of the new methods. Do you perhaps have any thoughts on how this could be improved, considering different possibilities (e.g. within BCH and Secretariat of the Convention on Biological Diversity). In case you have any thoughts on the role of regional networks in supporting knowledge sharing, I encourage you to contribute to Topic 4 of this discussion forum as well.

Kind regards,
David Dobnik

Dear Mr. Genetu,

Thank you for sharing your experience.
Considering different challenges, do you think they could be resolved in the future, so that you'll be able to establish the laboratory by implementing new technologies as well?

Kind regards,
David Dobnik

Sure, for the future all the challenges will be managed accordingly and we will go for the latest technologies.

Regards

Amare Genetu

I am Minosoa Barphyl ANDRIAMIHARISOA, a PhD candidate in Botany with a background in Plant Physiology and Biotechnology. Since 2021, I have been involved in LMO detection at the Plant Molecular Biology Laboratory of the University of Antananarivo (Madagascar), one of the two laboratories responsible for LMO detection in Madagascar. By participating in this online exchange, I hope to update my current knowledge on LMO detection, strengthen national capacity in this field, and share the country’s progress in LMO detection.

a) Are there any new analytical techniques (or protocols) that have been developed or implemented since 2023 that could be applied to detection and identification of LMOs? If so, please describe them?
In our laboratory, we currently do not employ any of the new analytical techniques for LMO detection. Our protocols rely primarily on RT-PCR and focus mainly on detecting the 35S and NOS genes. These analyses are performed only at the request of economic operators seeking to export products outside Madagascar, as the decree designating the two national laboratories responsible for GMO detection is still under validation. This regulatory situation significantly limits the scope of detection activities carried out in our laboratory.

b) Has the transition to the use of new methodologies been achieved in your laboratory?
Not yet.

c) Have these new methodologies been successfully implemented and has high throughput been achieved? If so, are the costs affordable? Are they part of the routine analysis? Has the number of targets increased?
Not yet.

d) If new methodologies have not yet been implemented in your laboratory, what factors have prevented their adoption, and what challenges do you anticipate in implementing them?
For Madagascar, the limited adoption of these new analytical techniques is mainly due to the lack of necessary materials and reagents for their implementation. In addition, insufficient capacity building hinders their integration, and, lastly, the absence of financial support from the national government for research further restricts their development.

Prof David,

I am happy to be part of this discussion and thank you for the initiative

We have not developed any new protocols since 2023 related to identification of LMOs. We mainly rely on the standard qPCR using commercially available kits. This, therefore implies that we have not transitioned to the use of the new methodologies as outlined by Dr. Freddy Bulubulu. The following factors may have contributed to the lack of our transition
1. Lack of access to the new tools and associated "specialized" equipment
2. Limited human capacity to apply the new methodologies
3. We sometimes have lengthy procurement procedures for laboratory supplies and equipment

Having said that, we strongly believe that we need robust tools in LMO detection to build confidence of our clients and therefore upgrading of our human, infrastructure and analytical capacity is vital

Dear David,
Thank you for the opportunity to contribute to this discussion.
My name is Dr. Belinda Akomeah, responding on behalf of the CSIR–CRI team.
These are our responses to Topic 1: Experiences and challenges in implementing new techniques for the detection and identification of LMOs

a) New analytical techniques developed or implemented since 2023.
We started our GMO detection work in 2023, and it involves an initial qualitative screen using conventional PCR, followed by real-time quantitative testing via qPCR (Bio-Rad GMO Investigator Kit) for positive samples. Since then, no new analytical techniques such as dPCR, NGS or LAMP-based assays have been developed or implemented beyond our existing GMO detection workflow.

b) Transition to new methodologies.
No, we haven’t tried any new methodologies yet, so the transition to new methods have not been achieved in our laboratory as we have not been able to adopt emerging techniques within our current operational framework.

c) Not applicable since new methodologies have not been implemented.

d) Factors or challenges preventing adoption and implementation.
Currently, we have not yet explored the inclusion of new methodologies or evaluated which technologies would be most appropriate to adopt, primarily due to:
1. Financial limitations: The cost of acquiring equipment for advanced methods, as well as the reagents, consumables, and certified reference materials required, poses a significant barrier in addition to resources needed for assay development, method validation, and participation in proficiency testing.
2. Limited training opportunities: While our laboratory has competent technical staff, we lack access to specialized training in emerging techniques such as digital PCR, next-generation sequencing, and other advanced molecular detection or quantification platforms.
We're glad to be part of this Network and keen to learn from the experiences from other laboratories in LMO detection

Dear Dr. David and colleagues,

My name is Maria da Glória Trindade, and I am an Agronomist specializing in Molecular Biology, Genetics, and Plant Breeding. I currently work as a Federal Inspector at the Federal Laboratory for Animal and Plant Health Inspection in Goiás (LFDA-GO), under the Ministry of Agriculture and Livestock of Brazil.
a. We are aware of new methodologies that can be applied to the detection and identification of LMOs, such as Next-Generation Sequencing, digital PCR, and other specific methods for genome-edited organisms, but only from the scientific literature. We do not yet have practical experience with these methodologies in the context of LMO analysis.
b. We have not yet transitioned to implementing new methodologies in our routine work.
c. We have tested microchip-based digital PCR, but we did not achieve good results in terms of cost–benefit, as the consumables are very expensive. Therefore, all methods currently used in our laboratory remain based on real-time PCR with primers and probes. Regarding NGS, although we use this technology for other purposes in our laboratory, the main challenge in applying it to LMO analysis is data analysis, particularly due to the lack of bioinformatics expertise in our unit. Additionally, to date, we have not encountered any scenario that we could not resolve using traditional methodologies. Over the past two years, we have experienced a significant increase in the number of targets analyzed in the laboratory, due to the approval of new LMOs in Brazil.
d. For most new techniques, we would need to invest in capacity building, as well as in acquiring new equipment and consumables that are not currently used in our laboratory. We are aware that the field of LMO detection and identification is evolving rapidly, and we need to be prepared for upcoming scenarios. We have taken some initial steps in this direction but still need to advance further.

Dear David,
Thank you for the opportunity to contribute to this On-line discussion.
My name is Dr. Wonkyun Choi, working at National Institute of Ecology in South Korea.
Since last 10 years, I have scientific experience for the development of LMO detection method using conventional PCR, LAMP and Lateral flow assay.

Our responses to Topic 1 is below

a) New analytical techniques developed or implemented since 2023.
We developed LMO detection method using conventional PCR, therefore, not new analytical techniques.
b) No, we haven’t considered any new methodologies yet.
c) N/A
d) The reasons of Not Implementing New Methodologies is the cost of acquiring equipment for new analytical techniques, especially high cost for set up the analytical equipment.

Dear Professor Dobnik and Colleagues,
Thank you for initiating this crucial discussion. My name is Danial Kahrizi, and for over 20 years, my research in Iran has focused on oilseed plants, particularly Camelina sativa. In our laboratory, the detection of LMOs/GMOs has traditionally relied on conventional PCR methods. While we also have the foundational expertise to perform CRISPR-based techniques in our research, our routine diagnostic pipeline for GMO detection has not yet fully integrated the latest advancements.
In response to your questions:
a) New techniques developed since 2023:
Based on the scientific literature, there have been significant developments since 2023 that are highly relevant to our field. The most notable advancements include:
1. Advanced PCR/dPCR for NGTs: There has been progress in developing event-specific qPCR assays for genome-edited plants (e.g., CRISPR-edited soybeans with deletions in the FAD2 genes). These methods use clever designs, such as primers overlapping the deletion site and incorporating LNA (Locked Nucleic Acid) bases, to precisely discriminate between edited and wild-type alleles. Furthermore, multiplex droplet digital PCR (ddPCR) assays have been validated for simultaneously quantifying multiple GM events in a single reaction with high precision, reducing both time and cost for screening.
2. Next-Generation Sequencing (NGS): The inter-laboratory reliability of NGS methods, especially hybridization probe capture followed by sequencing, has been demonstrated for detecting unknown or unauthorized GMOs, including those derived from New Genomic Techniques (NGTs). These are now being recognized as powerful complementary tools for comprehensive analysis.
3. CRISPR-Based Biosensors for Rapid Detection: Perhaps the most striking innovation is the development of portable, rapid detection systems. These combine isothermal amplification (like RPA or LAMP) with CRISPR/Cas12a proteins and a visual readout on a lateral flow strip. These assays can detect common transgenic elements (e.g., CP4-EPSPS, Cry1Ab/Ac) in under 45 minutes, offering immense potential for field-side or port-of-entry screening.
d) Challenges in implementation:
Regarding the factors preventing the adoption of these new methodologies in our laboratory, we face a unique and significant challenge: economic and technological sanctions.
Our country, Iran, is subject to extensive economic sanctions from the United States, which have recently been compounded by international restrictions. These sanctions directly impede our ability to acquire state-of-the-art equipment, reagents, and validated kits. Procuring advanced platforms like modern digital PCR systems or NGS hardware is often impossible.
To navigate this constraint, we have adopted a strategy of "knowledge transfer through human capital." We routinely send our top Ph.D. students abroad for research fellowships to leading institutions in countries that are at the forefront of this technology. There, they gain hands-on experience with these advanced assays and bring that invaluable expertise back to share with our team. While this is a suboptimal and costly workaround, it is a necessary one to ensure we do not fall completely behind the global scientific curve.
We are eager to learn from the experiences of other laboratories and are always open to collaborative efforts that can help bridge these technological gaps.
Sincerely,
Danial Kahrizi

Dear Professor Dobnik, Dear Colleagues,

Thank you to Professor Dobnik for framing this critical discussion and to Mr. Freddy Bulubulu Otono for his excellent, comprehensive review of the cutting-edge methodologies emerging since 2023.

My name is Danial Kahrizi, and for over two decades, my research in Iran has centered on the genomics and biotechnology of oilseed crops, with a dedicated focus on *Camelina sativa*. In our laboratory, we maintain a robust diagnostic pipeline based on conventional and qPCR methods, complemented by active research into CRISPR-based applications for plant engineering.

Mr. Otono's summary perfectly captures the technological landscape. I would like to build upon his points by adding a strategic perspective on the implementation hierarchy of these technologies, which may be of value to this network.

The advancements he outlined can be viewed not just as a list, but as a portfolio of tools for different tiers of the analytical workflow:

1.  CRISPR-Cas Biosensors for Tier-1 Screening: The developments in RPA/LAMP-CRISPR-LFA, as highlighted, represent a paradigm shift. Their true value lies in decentralized, rapid prescreening. This can act as a powerful triage mechanism at ports of entry or for initial field samples, potentially reducing the load on central, high-throughput laboratories by filtering out negative samples with high confidence. The challenge remains the development of validated, commercialized kits for a broader range of traits.

2.  Advanced dPCR for Tier-2 Quantification and Validation: The progress in multiplex ddPCR is significant for reference laboratories. Its strength in absolute quantification and tolerance to PCR inhibitors makes it the gold-standard successor to qPCR for precise event-specific quantification and for validating the results of rapid screens. Its role in providing definitive proof for regulatory compliance cannot be overstated.

3.  NGS as the Tier-3 Discovery and Forensic Platform: As Mr. Otono noted, NGS is our most powerful tool for hypothesis-free detection. Its primary implementation, in our view, should be targeted at several critical applications:
    *   Forensic Analysis of unauthorized or unknown GMOs/LMOs.
    *   Characterization of Complex Stacked Events.
    *   Comprehensive Safety Assessment of NGT products, where confirming the absence of off-target edits and residual vector sequences is paramount, as mentioned in the Singh et al. (2025) study on edited tomatoes.

Regarding implementation challenges, our situation in Iran presents a case study in navigating severe technological constraints. As a nation under extensive international sanctions, our primary impediment is systemic isolation from the global technological ecosystem. This manifests as:

*   An inability to procure state-of-the-art instrumentation and, critically, the proprietary reagents and validated kits required for their operation.
*   A critical gap in technical capacity building, as hands-on training on these platforms is inaccessible domestically.

Our mitigation strategy has been one of "intellectual arbitrage." We systematically deploy our most promising Ph.D. researchers to internationally leading laboratories for extended research fellowships. Their mission is to acquire not just theoretical knowledge, but practical, hands-on proficiency with these advanced platforms—from multiplex ddPCR to NGS library preparation and CRISPR-based assay development. They return as conduits of cutting-edge expertise, enabling us to maintain a baseline of competency and to creatively adapt existing in-house technologies to approximate new methodologies where possible.

In conclusion, the scientific roadmap is clear, as Mr. Otono has eloquently shown. The paramount challenge for many of us is no longer the *what*, but the *how*—how to bridge the profound implementation gap in the face of geopolitical and economic barriers. I believe this network is an ideal platform to foster not only the exchange of scientific knowledge but also innovative strategies for collaborative capacity building that can transcend these limitations.

I look forward to a fruitful discussion.

Sincerely,

Danial Kahrizi

Dear Professor Dobnik, Dear Mr. Genetu, and Colleagues,
Thank you, Mr. Genetu, for your candid and highly valuable contribution. Your description of the foundational challenges in establishing a GMO detection laboratory resonates profoundly and highlights a critical reality for many of us operating outside well-funded, long-established networks. Your perseverance in reorganizing and commencing testing under such constraints is commendable.
Your experience underscores a pivotal concept in our field: the "implementation chasm." This is the significant gap between the rapid pace of methodological innovation—as detailed excellently by Mr. Otono—and the practical, on-the-ground capacity to deploy even foundational technologies like qPCR.
Building on your points, I would like to emphasize that the challenges you listed are not isolated but are interconnected systemic barriers:
1. The Procurement Paradox: The difficulty in procuring high-quality instruments through national systems creates a vicious cycle. Without reliable equipment, generating the robust, accreditation-ready data required to justify further investment becomes an uphill battle. This often sidelines laboratories before they can even begin to demonstrate their potential value to regulators and funders.
2. The Capacity-Building Gap: The "Lack of Training" you mention is not merely about technical skills; it encompasses the entire ecosystem of method validation, quality control, and data interpretation. Establishing competence in conventional PCR is a necessary and commendable first step. The transition to qPCR/dPCR then represents a next-tier challenge, requiring not just a new instrument, but a shift towards absolute quantification, complex data analysis, and more stringent contamination control.
From our perspective in Iran, we face a parallel, albeit distinct, set of constraints due to comprehensive international sanctions, which mirror your access limitations. Our strategy, therefore, may be of mutual interest. We have focused on "knowledge repatriation" as a stopgap measure. By strategically sending our doctoral candidates to international partner laboratories, we aim to internalize the expertise that we cannot yet import in the form of hardware. This allows us to build a reservoir of human capital ready to deploy advanced methodologies the moment infrastructural barriers are lowered.
Mr. Genetu, your laboratory's current phase of building competency with conventional PCR is, in many ways, the most critical one. A solid, well-understood foundational system is the essential platform upon which all future advancements must be built.
Perhaps this network could explore tangible initiatives to bridge this "implementation chasm," such as:
• Creating a repository of validated, low-cost SOPs for conventional methods.
• Establishing a virtual "peer-mentoring" program where more experienced laboratories can provide remote guidance on method troubleshooting and validation.
• Collaboratively advocating at an international level for streamlined equipment donation or cost-sharing programs for nations facing severe procurement challenges.
Your voice in this discussion is vital, as it ensures that the conversation remains grounded in the realities of a global community striving for scientific excellence under unequal circumstances. I look forward to finding collaborative pathways forward.
Sincerely,
Danial Kahrizi

Dear Professor Dobnik, Dear Mr. Bulubulu Otono,

Thank you, Professor Dobnik, for steering this critical discussion towards actionable solutions. The challenge articulated by Mr. Bulubulu Otono is one that resonates deeply with my own laboratory's experience in Iran.

Mr. Otono, your comprehensive summary of the technological landscape stands in stark contrast to the implementation barriers we collectively face. In response to Professor Dobnik's pertinent question on how to bridge this gap, I would like to propose a multi-tiered strategy where the BCH and regional networks could play a transformative role:

1. The Role of the BCH and Secretariat: Orchestrating a "Knowledge and Technology Transfer Pipeline"
The BCH is uniquely positioned to move beyond being a repository of information and become a dynamic facilitator. Concrete actions could include:

    Establishing a "Validated Methods Vanguard" Program: A curated, peer-reviewed collection of SOPs for both foundational (qPCR) and advanced (ddPCR, CRISPR-LFA) techniques, specifically validated for use with cost-effective or open-source hardware platforms where possible.

    Creating a "Technology Access Fund": A mechanism, potentially supported by the Global Environment Facility (GEF) or other donors, specifically designed to co-finance the procurement of essential core equipment for qualifying national laboratories. This would directly address the procurement paradox that cripples nascent labs.

    Facilitating "Reagent Consortiums": Leveraging the collective purchasing power of multiple national laboratories through the Secretariat to negotiate favorable pricing and ensure reliable supply chains for critical reagents, bypassing inefficient national procurement systems.

2. The Power of Regional Networks: Building "Centers of Gravitas"
Regional collaboration is not just beneficial; it is essential for sustainability. We should aim to create:

    Regional Hubs for Technical Proficiency: Designating existing, well-equipped laboratories in a region as "Centers of Excellence" for hands-on, practical training. This is far more cost-effective and culturally relevant than sending individual researchers across the globe. These hubs could offer standardized courses on method validation, data analysis, and laboratory quality assurance.

    A "Floating Expertise" Model: Encouraging short-term, targeted secondments of experts from established laboratories (e.g., in Slovenia) to regional hubs, maximizing knowledge transfer impact.

    Shared Instrumentation Facilities: For high-cost, low-frequency-use equipment like NGS platforms, a regional shared-access model, managed through a central hub, could make this powerful technology accessible for forensic analysis and method development without each nation bearing the full capital cost.

In essence, we must transition from a model of isolated laboratories struggling independently to an integrated ecosystem of shared resources, standardized knowledge, and collaborative problem-solving.

Mr. Otono, your laboratory's deep theoretical understanding of the field, as demonstrated in your post, is the most valuable asset. Pairing that knowledge with the structural support of the BCH and the practical solidarity of a regional network is the most promising pathway forward.

I fully endorse Professor Dobnik's suggestion to continue this vital conversation under Topic 4, and I commit to contributing there as well.

Sincerely,

Danial Kahrizi

Dear Mr. Dobnik,
It is a great honour to be part of this discussion. Though I must mention that it is the first time interacting on this forum and it seems this is a review of earlier discussions from before regarding the specific questions you have posed.
Nonetheless, to address your specific questions:

a) There have been no new developed analytical techniques and our lab at the National institute for scientific and industrial research-Zambia relies  on old techniques and uses a conventional thermocycler for the analysis of LMOs/GMOs.

b) Regarding Transitioning to new Technologies, our lab has not done so.
c) No new methods have been implemented and no high throughput achieved
d) several factors have led to the non implementation of new technologies, chiefly being lack of modern equipment (the lab lags in technological advances and although there have been plans to equip in with modern, more efficient equipment, no equipment has been procured. Secondly, lack of any training over the same has not been there.

My name is Dr Francis Djankpa, I am a senior Lecturer in the University of Cape Coast (Ghana) and also a member of the Technical Advisory Committee (TAC) of the National Biosafety Authority (NBA) in Ghana. Here in Ghana we are not exposed to the new methods of GMO detection. Our lab is equipped with a conventional PCR machine which we use for GMO detection. We also have access to a qPCR for quantification as needed. Apart from these basic equipment, we don't have any advanced equipment for SGS, CRISPR systems for GMO detection. However, with the current detection methods that we are using, we have few challenges:

1. Access to reagents: Currently the only company that sells reagents for us in Ghana for GMO detection is based in South Africa and the minimum shipping time for delivery of reagents to Ghana is about 3months i.e. 12 weeks. Sometimes we could wait up to 6 months before receiving the reagents and this causes significant delays in operation.

2. High cost of reagents: The reagents are very expensive in South Africa compared to Europe and Asia but the companies in Europe and Asia say they cannot deliver directly to Ghana. So we have to continue to buy the reagents at a higher cost.

3. Primer information: Most of the companies that sell primers to us from South Africa are not willing to disclose the primer information to us. I am not sure why this is so......

4. General Laboratory Protocol Challenges: As expected, we have optimization challenges just like every other lab but we persist and do adjustments until we obtain the expected results.

Dear colleagues,

my name is Daniela Wahler. I work for the German Federal Office of Consumer Protection and Food Safety (BVL) in the Unit Methods of Detection, GMO Findings, Biosafety Clearing-House, EUginius. My Unit is home to the competent national authority for the Cartagena Protocol and also to a working group according to the German Genetic Engineering Act that has the legal mandate to publish an official collection of methods of sampling and analysis of LMO in Germany (§ 28b GenTG Working Group). I would like to thank the Secretariat for the opportunity to discuss here.

Implementing new techniques, i.e. advanced analytical methods, in official control is a long road. E.g., in Germany, digital PCR to be used for the detection and identification of LMO has been firstly explored more than 10 years ago. Although proving advantageous for quantification, digital PCR is not yet established in official controls. In Germany, the official control laboratories for the detection and identification of LMO so far use digital PCR for proficiency testing and adjusting samples and reference materials. However, digital PCR is further explored for its applicability in the detection and identification of LMOs and first attempts for standardisation have been successful (e.g. Overview and recommendations for the application of digital PCR, European Network of GMO Laboratories (ENGL), doi:10.2760/192883; Gatto et l., Definition of Minimum Performance Requirements for Analytical Methods of GMO Testing – part 2, doi:10.2760/63656; Arbeitsgruppe § 64 LFGB, Leitlinien zur Verifizierung von Methoden mittels digitaler PCR, https://www.bvl.bund.de/SharedDocs/Downloads/07_Untersuchungen/Leitlinien_Verifizierung_Methoden_digitale_PCR.pdf?__blob=publicationFile&v=12)

From my perspective, standardisation of next generation sequencing (NGS) will be a similar lengthy process.

Standardisation of new techniques is one essential step if a new technique is considered to be implemented in the detection and identification of LMO. But, as mentioned in previous posts (#12829, #12830, #12837), the integration of new techniques such as digital PCR and NGS in official controls requires financial resources, access to equipment and consumables, and well-trained laboratory staff. Both, digital PCR and NGS, are still more expensive than “classical” real-time PCR (commonly called qPCR), and trained staff is needed to achieve reliable analysis results when using new techniques. It should therefore be emphasised that the techniques currently established, mostly qPCR, are already very well-suited for detection purposes. For many standard tasks in detection, focusing on reliably applying these established techniques can already go a long way.

That is why, in my opinion, when integrating new techniques for the detection and identification of LMO, it is important to carefully consider the objective of the analysis.
- Is it a test for the presence of (unauthorized) GMOs? In most cases, qPCR is sufficient. It delivers a clear yes/no answer and is still most cost efficient.
- Is quantification necessary or required (e. g. labelling thresholds)? Digital PCR is a precise tool for quantification as no control samples or standards need to be included in the run and the method is more precise.
- Has an unknown LMO been detected in a sample that – for any reason – needs further molecular characterisation? Then the use of new NGS techniques may be the only option.

Please, also refer to my post on topic 4 on the role of networks where I want to state that not every laboratory needs to have the capability to implement every technique but networks with single specialized labs might be advantageous.

Kind regards,
Daniela

Dear Mr. Genetu,

Thank you for your positive and forward-looking note. Your optimism is both welcome and essential as we navigate these complex challenges.

I am glad to hear of your commitment to adopting the latest technologies. This is a goal we all share. As we move forward, the collaborative frameworks we are discussing here—such as regional knowledge-sharing initiatives and potential support mechanisms through the BCH—could prove invaluable in transforming that commitment into tangible reality.

Perhaps we can explore specific, collaborative projects in the future. For instance, a comparative study on the application of conventional PCR for specific traits in our respective regions could be a valuable first step in building a shared data foundation.

I look forward to our continued dialogue and potential collaboration.

Best regards,

Danial Kahrizi

Dear Ms. Andriamiharisoa, Dear Colleagues,

Thank you, Ms. Andriamiharisoa, for your highly insightful and candid contribution. Your description of the situation in Madagascar provides a crucial case study that underscores a fundamental truth: the challenge of implementing advanced detection methodologies is not merely technical, but is deeply intertwined with regulatory frameworks, institutional support, and sustainable funding models.

Your point about the pending national decree is particularly poignant. It highlights the "regulatory Catch-22" that many developing laboratories face: without a formal mandate, it is difficult to secure the funding and institutional buy-in needed to build capacity; and without demonstrated capacity, the urgency to finalize such mandates can remain low. Breaking this cycle is essential.

Your experience strongly resonates with the constraints mentioned by Mr. Otono and Mr. Genetu. It appears we are confronting a common triad of barriers:

The Infrastructure Gap: Lack of access to specialized reagents and equipment.

The Human Capital Gap: Insufficient targeted training on advanced platforms.

The Policy-Support Gap: A lack of synchronized national strategy that aligns regulatory needs with research funding.

In light of these shared challenges, the role of this Network becomes even more critical. Perhaps we can collectively advocate for mechanisms that address these gaps simultaneously. For instance, a potential action item for this community could be to draft a model "National Framework for LMO Detection Capacity" document. This could serve as a template for scientists like yourself to present to your national policymakers, outlining the necessary steps for regulatory validation, laboratory designation, and sustained funding.

Furthermore, your laboratory's existing proficiency with RT-PCR for the 35S and NOS promoters is a solid foundation. This core competency is the perfect platform upon which to build. A logical and cost-effective next step could be the implementation of a multiplex qPCR system. This would allow you to expand your target range (e.g., to include P35S, T-nos, pat, bar, and species-specific reference genes) in a single, more efficient reaction, directly enhancing the value of the services you provide to economic operators.

Ms. Andriamiharisoa, your proactive engagement as a Ph.D. candidate is exactly the kind of initiative that will drive change in Madagascar and beyond. I encourage you to view this forum not only as a source of knowledge but as a potential platform for building the collaborative partnerships—perhaps with neighboring countries in the African region—that are so vital for overcoming our shared limitations.

I am keen to explore how we can translate this discussion into concrete, collaborative actions to support laboratories like yours.

Sincerely,

Danial Kahrizi

Dear Mr. Mtonga, Dear Colleagues,

Thank you, Mr. Mtonga, for your valuable contribution, which adds another critical data point to our evolving understanding of the global landscape for LMO detection. Your point about the need for "robust tools to build client confidence" is absolutely fundamental and gets to the very heart of why our work matters. This confidence is the cornerstone of trade, biosafety, and evidence-based regulation.

Your summary of the contributing factors—access, capacity, and procurement—perfectly encapsulates the core triad of constraints we have been discussing. It is increasingly clear that these are not isolated national issues but a common syndrome affecting many laboratories dedicated to this crucial field.

I would like to build upon your astute observation by suggesting that the solution lies in addressing these three gaps not in isolation, but through an integrated approach. Perhaps we, as a network, can begin to conceptualize a "Capacity Building Triad" model:

For Access (Tools & Equipment): We must explore innovative procurement models. As I mentioned previously, this could include regional equipment-sharing consortia for high-end instruments and leveraging the BCH's potential to facilitate bulk reagent purchasing to overcome cost and logistical barriers.

For Capacity (Human Expertise): The "limited human capacity" you cite is a bottleneck we all recognize. Beyond one-off trainings, we should advocate for the establishment of a "Twinning Program" within this network, pairing experienced laboratories with those in the development phase for sustained mentorship, remote troubleshooting, and collaborative method validation.

For Procurement (Administrative Efficiency): The challenge of "lengthy procurement procedures" is a systemic one. A potential role for the Secretariat could be to develop and disseminate a "Model Procurement Guide" for essential LMO detection technologies. This guide would provide standard technical specifications, aiding national laboratories in streamlining their own procurement justifications and processes.

Mr. Mtonga, your laboratory's continued operation with standard qPCR demonstrates resilience. This existing platform is your strategic asset. The transition to some of the new methodologies, such as the multiplex ddPCR assays mentioned by Mr. Otono, can be viewed as a natural evolution from your current qPCR work, offering enhanced robustness and multiplexing capabilities that directly address your goal of building client confidence through more powerful data.

Your commitment to upgrading capacity, even while facing these headwinds, is precisely the mindset that will drive progress. I am confident that by continuing to articulate these shared challenges and collaboratively proposing concrete solutions, we can build a compelling case for targeted international support.

I look forward to working with you and others in this forum to translate this "Triad" from a concept into a practical action plan.

Sincerely,

Danial Kahrizi

Dear Dr. Akomeah and the CSIR–CRI Team,

Thank you for your clear and concise contribution, which provides a highly valuable perspective from a laboratory in the early and crucial stages of establishing its LMO detection capabilities.

Your implementation of a two-tiered workflow—using conventional PCR for cost-effective screening followed by event-specific qPCR for quantification—is a highly strategic and commendable approach. This methodology represents a robust foundation that prioritizes reliability and regulatory compliance, which are essential for building the very "client confidence" that Mr. Mtonga rightly emphasized.

Your identification of financial limitations and training access as the primary barriers is precise and resonates deeply with the collective experience in this forum. It is particularly noteworthy that you highlight the often-overlooked but critical costs of certified reference materials and proficiency testing, which are non-negotiable for generating internationally recognized data.

Your situation presents a unique opportunity for a targeted and phased strategy for capacity building. Rather than a disruptive leap to advanced platforms, your logical next step could be a focused expansion within your existing qPCR framework:

Phased Expansion of qPCR Capabilities: Before considering dPCR or NGS, a strategic and cost-effective evolution would be to develop in-house multiplex qPCR assays. This would allow you to expand your target range (e.g., to include common genetic elements like P-35S, T-NOS, pat/bar, and species-specific genes) within a single reaction, thereby increasing throughput and reducing per-test costs without requiring entirely new instrumentation.

A Gateway to Rapid Testing: Furthermore, the expertise your team is building with PCR is directly transferable to evaluating CRISPR-Cas12a based lateral flow assays. These rapid tests could serve as an even faster and more economical pre-screening tool than your conventional PCR, potentially freeing up your qPCR system for more complex quantification tasks.

Dr. Akomeah, your team's keenness to learn is your greatest asset. This network should actively explore mechanisms to support laboratories like yours, such as establishing a digital repository of low-cost, validated SOPs for in-house assay development or facilitating virtual "train-the-trainer" sessions on specific topics like multiplex assay design and validation.

The foundational work you are doing is critical. By strengthening your current systems and strategically planning your technological evolution, you are building a sustainable path toward future advancements. We are all here to learn from each other, and your proactive engagement is a vital part of that process.

I look forward to our continued exchange.

Sincerely,

Danial Kahrizi

Dear Ms. Trindade,

Thank you for your exceptionally insightful contribution. Your perspective from a high-throughput federal laboratory in Brazil is incredibly valuable, as it highlights a critical evolutionary stage in LMO detection: the transition from a well-established, highly effective qPCR system to next-generation technologies, not out of necessity, but for future-proofing and gaining a strategic advantage.

Your point regarding the "cost-benefit" analysis of dPCR is a crucial reality check for the entire community. It underscores that economic viability is as important as technical performance for the routine implementation of any new method.

Furthermore, you have pinpointed perhaps the most significant bottleneck for applying NGS in our field: the bioinformatics gap. The statement, "the main challenge... is data analysis, particularly due to the lack of bioinformatics expertise," resonates profoundly. Many laboratories can generate the sequencing data, but the expertise to transform millions of reads into a clear, actionable result—such as identifying an unknown GMO or confirming a complex edit—is a specialized skill set that is not yet widespread.

In this context, and to directly address the collaborative spirit of this network, I would like to propose a concrete offer of cooperation.

My laboratory in Iran has, out of necessity, developed a strong in-house capacity for the full pipeline of NGS data analysis, from raw read processing and quality control to advanced bioinformatic analyses for the detection of unknown genetic modifications, vector backbone analysis, and off-target assessment in NGT products.

We would be very interested in exploring a collaborative partnership with your laboratory. A potential pilot project could involve your team generating the NGS data from a challenging sample and our team handling the comprehensive bioinformatic analysis. This would allow you to evaluate the power of NGS for LMO detection without the immediate need for a large internal investment in bioinformatics training and staffing.

We are confident that such a collaboration could be mutually beneficial, providing you with valuable data and insights while allowing us to apply our expertise to real-world scenarios from a leading agricultural nation.

Please feel free to contact me directly to discuss this possibility further.

Mobile: +98 918 332 2235
Email: dkahrizi@modares.ac.ir

We look forward to the possibility of working together to bridge the bioinformatics gap and unlock the full potential of NGS for the global LMO detection community.

Sincerely,

Danial Kahrizi

Dear Dr. Choi,

Thank you for your valuable contribution. It is insightful to hear from a colleague with a solid decade of experience in developing detection methods, including practical approaches like LAMP and Lateral Flow Assays (LFA). Your work on these platforms is highly relevant, especially for rapid on-site screening applications.

Your point regarding the prohibitive cost of acquiring and setting up new analytical equipment is a fundamental challenge that resonates across many laboratories, regardless of their national context. It underscores the importance of a cost-benefit analysis for any technological transition.

Given your extensive expertise in LFA and LAMP, I see a significant opportunity for synergy. My laboratory in Iran has developed a strong focus on adapting CRISPR-Cas based systems for rapid diagnostics. The principles are complementary to your work with LFA.

I would like to propose a potential collaboration. A compelling and cost-effective research project could involve developing a novel, multiplexed CRISPR-LFA for a specific LMO target, leveraging your hands-on experience with lateral flow platforms and our work on CRISPR assay design. Such a collaboration would not require substantial investment in new equipment but would instead build upon both our strengths to create a next-generation rapid test.

This could be a fruitful way to innovate without the immediate financial burden of acquiring major new platforms like dPCR or NGS.

We would be very interested in exploring this possibility further.

Mobile: +98 918 332 2235
Email: dkahrizi@modares.ac.ir

I look forward to the possibility of combining our expertise.

Sincerely,

Danial Kahrizi

Dear Mr. Mulubwa,

Welcome to this forum. Your voice is a vital and welcome addition, as it highlights the foundational challenges that must be addressed to build a truly global capacity for LMO detection. Thank you for your candidness in describing the situation at your laboratory.

The constraints you face—the lack of modern equipment and the complete absence of targeted training—represent the most fundamental barriers to entry in our field. It is precisely these gaps that this international network is best positioned to address through collaboration and direct knowledge transfer.

In line with this, I would like to extend a concrete offer of support. My laboratory actively hosts international researchers at both the graduate (MSc/PhD) and postdoctoral levels.

We have established, cost-effective workflows for core molecular techniques and are developing innovative approaches for LMO detection. Hosting a researcher from your institution for a focused training period could be a powerful strategy to bypass the current limitations.

A visiting scholar could gain hands-on experience with:

Advanced PCR techniques (qPCR, multiplex assay design).

Method validation and data analysis.

The principles behind rapid detection kits (LAMP, Lateral Flow).

This "capacity-building through immersion" approach would equip them with the practical skills and knowledge to become a catalyst for modernization upon their return to Zambia, effectively building your in-house human capital even before new equipment is procured.

We would be honored to host a dedicated candidate from your team and collaborate in building a sustainable future for LMO detection in Zambia.

Please feel free to contact me directly to explore this possibility further.

Mobile: +98 918 332 2235
Email: dkahrizi@modares.ac.ir

Sincerely,

Danial Kahrizi

Dear Dr. Djankpa,

Thank you for your profoundly insightful and critically important contribution. You have moved beyond listing general barriers and pinpointed the severe operational and logistical strangleholds that can paralyze even a well-equipped laboratory. The supply chain issues you describe are not just inconveniences; they are fundamental threats to the sustainability of national biosafety monitoring.

The challenges you outline—the 3-6 month reagent delays, exorbitant costs due to regional markups, and the unacceptable withholding of primer sequences—paint a clear picture of a non-competitive and restrictive supply environment. This situation is untenable for a National Biosafety Authority that requires timely and reliable data.

In direct response to these challenges, I would like to propose several concrete actions where international collaboration within this network could provide immediate relief:

Establishing a Reagent & Primer Consortium: This forum, perhaps through the Secretariat, could facilitate the creation of a collective purchasing group for member laboratories. By aggregating our orders, we could negotiate directly with European or Asian manufacturers for bulk purchases and reliable shipping, bypassing the costly middlemen.

Open-Source Primer Sequence Repository: To break the dependency on uncooperative suppliers, we could initiate a collaborative, open-access database within this network where validated primer and probe sequences for common GMO/LMO events are shared. This would empower laboratories to order from any synthesis company globally, fostering competition, reducing costs, and ensuring transparency.

Direct Collaboration to Bypass Barriers: My laboratory would be willing to act as a logistical hub for your time-sensitive reagent needs on a case-by-case basis. We could procure and expedite shipments to you, significantly reducing the wait time. Furthermore, we have extensive experience in in-house primer and probe design and validation. We would be pleased to collaborate with you directly to design your own assays, freeing you from the companies that withhold sequence information.

Dr. Djankpa, your persistence in the face of these obstacles is commendable. Let us use the collective power of this network to tackle these systemic issues head-on. I am ready to work with you and the Secretariat to transform these proposals into reality.

Mobile: +98 918 332 2235
Email: dkahrizi@modares.ac.ir

Sincerely,

Danial Kahrizi

Dear Ms. Trindade,

Thank you for providing your views.

I would like to comment on the use of digital PCR in terms of cost-benefit. As we have been using and developing dPCR methods since 2012, we have performed also a lot of cost evaluations, especially considering the comparison to qPCR. In general dPCR is more expensive than qPCR and our hopes to see the decrease of reagent prices over time have not realized. Therefore our initial idea was to explore the multiplexing capabilities and we have seen that this can move the costs closer to the qPCR, but it was limited to specific scenarios of having positive samples suitable for quantification. With the emergence of many LMO lines that lack the common screening elements, the qPCR analysis has also become more laborious and more costly as a lot of additional identification reactions need to be performed in parallel to screening. In fact for GM soybean lines we usually just perform the analysis by prespotted plates, analyzing all GM soybean lines in one plate (up to 3 samples per run). This has led us to revisit the multiplexing dPCR strategy and with availability of dPCR platforms with several fluorescence channels, this was a bit easier than in the past. We have just published a protocol for quantifiaction of EU approved GM soybean lines (https://experiments.springernature.com/articles/10.1007/978-1-0716-4767-7_2). We have also compared the costs of whole analysis and considering the analysis of 2 samples per run, the costs of multiplex dPCR and qPCR prespotted plate were the same. However, dPCR approach has one big benefit - in case any GM line is present, we will get the quantitative result at the same time. In the case of qPCR, separate analysis need to be performed for this purpose, which in the end increase the total costs of analysis. Thus multiplex dPCR really seems as more cost-effective option in this case.

Dear Mr. Kahrizi,

I would like to thank you for your valuable contributions so far.

After the closure of the form discussion, we will prepare a summary, including the proposals for improvement of LMO identification and detection in general. They could serve as a guidance or recommendations for any future actions, which might come from the CBD secretariat or perhaps any of the regional networks.
Therefore views and comments like yours are of great value for the field and I would like to encourage others to engage in the discussions as well.

Kind regards,
David

Dear Professor Dobnik,

Thank you very much for your kind and encouraging message. It is a genuine privilege to contribute to these critical discussions alongside such distinguished colleagues.

Hearing that our collective insights—including those regarding regional collaboration, reference material challenges, and strategic implementation of new technologies—may inform future guidance and recommendations is exceptionally rewarding. This is precisely the tangible outcome we all hope for when engaging in these forums.

The potential for our shared input to shape initiatives from the CBD Secretariat or regional networks is a powerful motivation. I firmly believe that the path toward robust, equitable, and globally harmonized LMO detection capabilities lies in the very kind of open, practical, and solution-oriented dialogue this Network fosters.

Please be assured of my continued and enthusiastic engagement. I remain fully committed to collaborating with you, the Secretariat, and all colleagues to translate these discussions into concrete actions that strengthen our global biosafety framework.

Thank you once again for your leadership in facilitating this immensely valuable exchange.

Sincerely,

Danial Kahrizi

Dear David,
Thank you for the opportunity to share my experiences in the GMO detection laboratory. Although we have the necessary equipment, the lack of specialized reagents remains a major challenge, as there are no suitable suppliers. In addition, we lack training opportunities to strengthen our capacities and adopt new technologies, which limits the effectiveness of our detection activities. To answer your questions, I would like to share the following points :
a) We have not yet reached this stage due to the lack of necessary reagents.
b) Our laboratory has not yet initiated the adoption of new technologies, mainly because of the lack of reagents and the absence of opportunities for specialized training.
c) We are in the process of starting analyses despite the challenge posed by the absence of local distributors of reagents. This situation results in high costs due to additional expenses related to the acquisition of reagents.
d) The main factors delaying the implementation of new technologies are the difficulty of accessing reagents and the lack of opportunities for specialized training.
Best regards
Mouhamadou MOUNKAILA BOUREIMA

Dear David,

Thank you for effectively moderating these discussions. Thank you Colleagues for your insightful engagements and the SCBD for facilitating.
My name is Josephine Amedu with the National Biosafety Management Agency, Nigeria. Our GMO Detection and Analysis Laboratory, established in 2017 employs protein based and PCR based approaches for detection. We recently strengthened capacity on qPCR (thanks to Herman Broll and Almut Leffke of the German Institute for Risk Assessment) and conducted a market survey for GM products and their compliance to labelling. Our published research article can be accessed at
https://doi.org/10.1016/j.jgeb.2025.100481.
In terms of implementing new detection techniques, such as dPCR and next generation sequencing we are not there yet.
Challenges encountered are similar to what our colleagues from Africa have cited, including lack of equipment and training on their use and application. This is quite worrisome, as these new techniques are viewed to be more applicable in detecting unauthorized GMOs and GMOs derived from emerging biotechnologies.
It is hoped that recommendations from these deliberations would help bridge these gaps identified.
Thank you.

Hello, we are Dr Hamidou MAIGA and Mr Inoussa TOE. We work together to answer questions on this platform as representatives of the team at the instititut de recherche en sciences - Direction Régionale de l'ouest de la santé (IRSS-DRO) of Burkina Faso.

a) New analytical techniques developed since 2023 and applicable to the detection/identification of LMOs

Over the past two years, several significant advances have been made in the field of genetic analysis of LMOs, including those derived from new genomic techniques (NGTs) or gene drive systems.

1. Increased use of eDNA (environmental DNA) to detect modified organisms
eDNA can be used to detect genetically modified organisms present in the environment, even at very low levels.
Growing application for monitoring released modified organisms, including gene drive systems.
Reference:
Piaggio A.J. et al. (2024). Building an eDNA surveillance toolkit for invasive rodents on islands: can we detect wild-type and gene drive Mus musculus? BMC Biology. https://doi.org/10.1186/s12915-024-02063-0
Direct application of eDNA to distinguish between wild individuals and individuals carrying a gene drive.

2. Expansion of ddPCR (digital droplet PCR) approaches for ultra-sensitive detection of GMOs and NGTs
ddPCR is becoming a key technology for low-abundance GMOs, CRISPR mutations without transgenes, and degraded matrices.
References:
• Wang K. et al. (2025). A Novel Quantification Method for Gene-Edited Animal Detection Based on ddPCR. Biology. DOI: https://doi.org/10.3390/biology14020203
• Fraiture M.-A. et al. (2022). ddPCR strategy to detect a gene-edited plant carrying a single base pair edit. Food Chemistry. DOI: https://doi.org/10.1016/j.foodcont.2022.108904
• Tigst Demeke Considerations for the Successful Detection and Quantification of Genetically Modified Events in Grain and Food Samples Using Multiplex Digital PCR. https://doi.org/10.3390/foods14010075

3. High-resolution multiplex qPCR for GMOs/NGTs
New multiplex qPCR  integrated into the European ENGL/EURL networks.
Targeting of specific promoters, terminators, transgenic junctions, and CRISPR signatures.
Reference:
• ENGL (2023). Detection of food and feed plant products obtained by targeted mutagenesis and cisgenesis. (Official European report, widely used for NGT screening.)
• Košir et al., 2023; Fast and Accurate Multiplex Identification and Quantification of Seven Genetically Modified Soybean Lines Using Six-Color Digital PCR. https://doi.org/10.3390/foods12224156
Gong Z. et al. (2025). Comprehensive benchmarking of genome editing quantification methods for plant applications. https://doi.org/10.1016/j.isci.2025.112350

b) The transition to the use of new methodologies has not been yet completed in my laboratory.

c) NA

d) We have solid expertise in bioinformatics, Sequencing, advances molecular biology and eDNA analysis, which puts us in a favourable position to tackle the technical challenges we typically encounter. However, our main challenge remains maintaining sustained financial investment to retain our technological lead in a rapidly evolving field.

Dear colleagues,

my name is Reona Takabatake. I work for the Institute of Food Research,
National Agriculture and Food Research Organization, NARO, Japan.
Our team has developed several GMO detection methods.
・ΔΔCq analysis for non-GMO labeling https://doi.org/10.1021/acs.analchem.2c02447
・LAMP-mediated detection method https://doi.org/10.1016/j.foodchem.2017.12.036
・Rapid and simple DNA chromatography-mediated detection method https://doi.org/10.1021/acs.jafc.8b01765
・Detection Method targeting an Ultrashort 25 bp Sequence for Agrobacterium-Mediated Transformed GM Plants https://doi.org/10.1021/acs.jafc.0c03864
In Japan, new technologies such as digital PCR or NGS has not yet established in official control. As others have noted, funding and equipment are essential to introducing these new technologies into public management. If these new technologies can solve problems that cannot be solved by qPCR, they may become more widely used. At least, I believe that NGS can be used to comprehensively detect unknown, unapproved GMOs.
We have used real-time PCR and LAMP method in combination to test imported seeds and seedlings. https://doi.org/10.1248/bpb.b22-00874
We have developed Pre-Spotted Plates (PSP), designated as real-time PCR array method, on a research purpose and used to identify GM cotton events.
https://doi.org/10.5740/jaoacint.11-388
In Japan, many GM events including 19 GM soy and 25 GM maize events have been approved, to improve the efficiency of inspections, the PSP method will be introduced to other major crops in the future.

I have a question. The number of samples subject to inspection varies greatly seasonally, and it seems that efficiency could be improved if screening inspections could be conducted using pool testing methods which is a method of grouping several samples to be analyzed together. Are there any countries that have introduced pool testing?

Dear Mr. Reona Takabatake,

Here in Brazil, we have not yet introduced pooled testing, but I believe this strategy could be useful in situations where mostly negative results are expected, as it can save both time and cost. However, if a pooled test produces a positive result, it is necessary to break the pool and test the individual samples. Possible applications here would include screening organic samples or testing species for which we have not approved LMOs.

Warm regards,
Maria

Dear Mr. Takabatake,

Thank you for sharing the developments of your team.

The pooling of the samples, which you have mentioned is definitely a good idea, especially for the samples, which are usually negative. As was mentioned by MSc Maria da Glória Trindade, any positive pool would mean additional testing of all individual samples. Our lab does not perform such kind of testing, but as we have several negative samples, this might be a good option to reduce the overal costs of LMO detection.

Kind regards,
David

Dear MSc Maria da Glória Trindade and Mr. David Dobnik, thank you very much for your message. As you mentioned, I think that pool testing will be effective when it is not expected to hardly obtain positive results. There may also be a problem with detection sensitivity. The risk of false negatives might increase as the sample is diluted by pooling. If actually pooled testing is introduced to GMO testing, it may be necessary to confirm in advance such things as what the detection limit and how many samples can be pooled.

Sincerely,

Reona Takabatake

Dear colleagues,
I am Dr. BODJONA Bawoumodom Pyabalo I Tchaou, head of the Crop Protection and Biosecurity Laboratory at the Togolese Institute of Agronomic Research (ITRA), located in Lomé Cacaveli. This laboratory is part of the ITRA and operates under the supervision of the Ministry of Agriculture. In Togo, GMOs fall under the umbrella of agricultural biosecurity, which is itself managed by the Ministry of the Environment and Forest Resources (MERF). This laboratory was equipped with the necessary equipment to conduct GMO detection analyses as part of a sub-regional program for the Prevention of Biotechnological Risks in West Africa, funded by the West African Economic and Monetary Union (UEMOA). The laboratory is involved in several areas, including GMO detection. This detection is performed on food products intended for export, such as soybeans, corn, etc.

For this GMO detection analysis, the laboratory has a choice between two methods. The first method is based on the use of conventional PCR, which is commonly used, and the second method is based on real-time PCR.
Since the beginning of 2025, the laboratory has decided to switch to real-time PCR for GMO detection; this technique, unlike conventional PCR, allows for quantification. The reagents have been acquired, and analyses will begin in January 2026.
However, the laboratory faces several challenges, including training new staff on the equipment (RT-PCR), implementing an interlaboratory comparison system to enhance the reliability of the analysis results, and obtaining ISO 17025 accreditation. Successfully meeting these challenges will allow the laboratory to provide reliable results to its clients.

We hope that these discussions will lead to funding opportunities that will allow the implementation of an inter-laboratory comparison system.

Thank you Mr. Kahrizi, I will definitely get in touch with you concerning your offered support, it would be a great honor to get the much needed help from you sir. regards.

Dear David:

Thank you very much for moderating this forum. My name is David Castro, and I am the Director of Genetic Resources and Biosecurity at the Ministry of Environment of Peru. I am sending my answers to the guiding questions.

To provide background, Peru currently has a single laboratory dedicated to the detection and identification of GMOs. This facility operates within the National Institute of Agrarian Innovation and holds accreditation under the ISO 17025 standard. It conducts testing using real-time PCR methodologies, including screening techniques such as p35S and tNOS, as well as certain event-specific assays.

a) To date, Peru has not adopted new detection platforms beyond established field tests and molecular confirmation. Routine environmental monitoring continues to rely on lateral flow test strips (Envirologix Quick Comb and Agdia single strips) for on-site screening and real-time PCR for laboratory confirmation. These methods meet current national regulatory objectives and have therefore remained the primary tools for detection and identification of LMOs.

b) There has been no full transition to novel methodologies in our laboratory. Because lateral flow screening combined with real-time PCR satisfies the legal and operational requirements at present, we have not moved to implement alternative technologies as standard practice.

c) In our country, adopting advanced platforms would likely face the following constraints:
- High initial investment for instruments plus ongoing expenses for brand-specific consumables and calibration. Many platforms require proprietary kits, preventing the use of lower‑cost generic reagents.
- Skilled personnel are needed to run and interpret advanced assays; training and retaining full time staff is costly if sample volumes are low.
- With many new LMO events emerging each year, single step methods that detect and identify multiple events across different matrices would be ideal; otherwise, the current two step approach (screening then event specific testing) becomes very expensive.

d) Funding must cover equipment purchase, maintenance contracts, calibration, and proprietary kits, considering low or variable demand for LMO testing may not justify hiring or retaining specialized staff full time.

Dear Dr. David,
My name is Yeny Aquino, head of the Laboratory for the Detection of Living Modified Organisms at the National Institute for Agricultural Innovation of Peru. Our answers are below:

a) We know that digital PCR is used for the quantification of GMOs, and we are interested in adopting this technique.
b) In our laboratory, we use real-time PCR for both the detection and quantification of GMOs.
c) We have not encountered any difficulties with the real-time PCR technique. With regard to costs, the reagents are expensive for us, but it is the import time that concerns us.
d) We have a limited budget, so it is not possible to buy equipment such as digital PCR.

Sincerely.

It´s very satisfying to share in this forum the experience we have gathered over more than 15 years in the detection of GMOs, the National Reference Center for Agrifood Safety and Biosecurity (CNRIBA), attached to the General Directorate of Agrifood, Aquaculture, and Fisheries Safety of the National Service for Agrifood Health, Safety, and Quality (SENASICA), is currently the only official laboratory in Mexico operating for the analysis of GMOs. It has been accredited since 2011 under the ISO-IEC-17025:2017 standard, provides training to colleagues from Latin American countries, and coordinates the Latin American and Caribbean Network for the Detection of Genetically Modified Organisms during the 2024-2025 period.

An area of opportunity identified is having greater access to scientific information (scientific articles) and training to implement new techniques. Additionally, support for equipment acquisition would be very beneficial, as budgetary policies make the acquisition process excessively lengthy.

One aspect that significantly impacts the center is the brain drain. Competitive in economic terms, once the trained technical staff is available, they obtain better job opportunities and leave the institution.

Hello everyone,
I represent the GMOs Testing Laboratory at the American University of Science and Technology (AUST) in Lebanon. Our laboratory was the first facility in the country to obtain ISO/IEC 17025 accreditation for GMO screening, identification, and quantification in seeds, food, and feed matrices, and we are currently undergoing accreditation renewal.
We implement ISO and EURL-recommended methodologies, relying primarily on Real-Time PCR and, in some cases, conventional PCR. The laboratory is well equipped, and our staff members are trained to conduct a broad range of molecular analyses relevant to LMO/GMO detection.
A key challenge we face in expanding and updating our detection capabilities is access to a comprehensive panel of primers, probes, and CRMs needed to cover a wide range of events. In Lebanon, GMO testing is not mandatory, resulting in a low and unpredictable volume of samples. This makes it difficult for us, as an academic institution, to continually invest in and maintain an extensive inventory of reagents, especially for events that may be rarely encountered.
To help address this gap, we are leveraging our academic mandate to develop research projects aimed at assessing the presence and status of GMOs and LMOs in the country. We believe that generating baseline data will support evidence-based national decision-making and strengthen our case for expanding testing capabilities.
We hope that this forum will contribute to shared solutions, including resource-efficient strategies, and collaborative access to reagents or reference materials.
Thank you

Good evening everyone.

My name is Galina Mozgova. I head the National Coordination Biosafety Centre of the Institute of Genetics and Cytology, with an ISO/IEC 17025 accredited laboratory specializing in the detection, identification, and quantification of LMOs/GMOs, and the determination of product counterfeiting using the method of species identification. Since such testing is mandatory in our country, it facilitates large-scale screening of incoming products and seeds.
We don't yet have experience with genome-edited organisms, but we do have scientific experience developing DNA markers for various types of mutations in agricultural crops, which is very similar and could facilitate work in the area of ​​detecting LMOs produced by genome-editing methods (RT PCR, DNA and RNA sequencing, SNaPshot analysis, PCR followed by restriction, and others). However, there are also challenges, such as the lack of reference materials or insufficient information about the edited regions of the genome of such LMOs. Such sequences aren't always added to databases. In addition to genetically engineered plants, the number of genetically engineered animals, including fish and mammals, is growing. These aren't typically detected in laboratories. I believe this is an important topic that requires collaboration, cooperation between countries, and capacity building.
Furthermore, the rapid development of new LMOs, including those produced through genome editing, requires time and resources to incorporate detection methods into existing large-scale screening schemes. I agree that scientific research in this area is important. This year, we are participating in a national competition for scientific projects to develop screening systems for unapproved LMOs with high potential for environmental invasiveness.

Best regards,

Galina Mozgova

Dear Dr. Danial Kahrizi,

Thank you very much for your kind words and for proposing a collaboration on NGS data analysis. I believe this potential collaboration could be very impactful in the future. However, so far all the NGS data generated in our laboratory has been related to metabarcoding based on the 16S rRNA and ITS regions. We do not currently have any data derived from LMO analysis.

I would like to thank you once again for your offer, and I look forward to continuing to explore collaboration opportunities that could be valuable for both our laboratories and our countries.

Kind regards,
Maria

Dear Dr. David,

Thank you for sharing your experience with dPCR, especially regarding multiplexing targets to help lower costs. We have not tested this approach in our laboratory yet. In fact, we are dealing with a complex variety of samples and targets, so we will need to think carefully about the most effective procedure to implement dPCR in our routine.

Thank you as well for sharing your paper with us. I will review it in depth to gain insights that we may be able to apply in our lab.

Best regards,
Maria

Dear Participants,

Thank you very much for your invaluable contributions and active participation in the Online discussions of the Network of Laboratories for the Detection and Identification of Living Modified Organisms. Your thoughtful insights and shared experiences have greatly enriched the exchange.
The Secretariat will carefully review and analyse the information provided and will prepare a comprehensive synthesis to inform and support future work of the network.

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Kind regards,
The Secretariat