Past Discussions 2013

Emerging Techniques for LMO Detection

Discussions will be held between 14 July - 28 July 2013
Discussions open and close at 1:00 a.m. GMT

Many of the techniques for LMO detection currently in use rely on well-established DNA- and protein-based methodologies, such as PCR-based assays, Southern blotting and protein expression analysis through ELISA and lateral flow assays.

During the previous discussions several participants indicated that they are actively undertaking research to fine tune and improve the methodologies that are currently in use in their laboratories. Furthermore the literature describes many new techniques, such as the use of microarrays, mass spectrometry and next generation DNA sequencing as well as the development of new software, which have the potential to be integrated into the routine workflows of LMO detection laboratories. Improved methodologies and emerging techniques may offer innovative and more cost-effective solutions to complex issues, such as the detection of adventitious LMO events and quantifying LMOs with stacked traits, and increased accuracy and efficiency to screen a greater number of samples.

In this discussion participants are invited to share information on what new technologies or methodologies are being researched or introduced in their laboratories, specifically:

1. Is your laboratory developing or has adopted novel technologies for the detection of LMOs? If so, which technologies are these and what benefits do they bring?
2. What techniques are laboratories researching to improve upon their existing protocols?
3. In their efforts to improve existing methodologies, what criteria are laboratories using to validate their new methodologies? Are there guidelines for the validation of new methodologies to assist laboratories in developing or adopting such methodologies?
4. What other technical barriers are faced by laboratories in their efforts to integrate new methodologies and how can they be overcome, for example resource and information sharing amongst national and/ or regional laboratories?

Challenges and Progress in the Detection of LMOs Unintentionally Released into the Environment and Unauthorized LMOs

Discussions will be held between 14 July - 28 July 2013
Discussions open and close at 1:00 a.m. GMT

The presence of an LMO in agricultural commodities that are authorized for use in a country but not yet approved in another country can occur through the inadvertent introduction of materials through standard manufacturing practices or unintended environmental release. While document based tractability of shipment contents can be used as a tool to track the presence of known and authorized LMOs, analytical methods to identify and quantify the presence of LMOs are essential in the detection of unauthorized LMOs and of LMOs unintentionally released, e.g., into the environment.

For the purposes of this discussion, an “unauthorized LMO” refers to the entry in the supply chain of an LMO that is either authorized for commercial use or sale in one or more countries, but is not yet authorized in an importing country (i.e. “asymmetric” or "asynchronous" authorisation). On the other hand, “unintentionally released LMO” refers to the introduction of LMOs which have not been assessed for use or release in any country, for example LMOs that are under research and development, and are often referred to as “unknown LMOs”.

Some laboratories are relying on a matrix-based approach to detect unintended and unauthorized LMOs. In this approach, samples are screened for the presence of genetic elements or constructs and the data obtained from such screenings can then be assessed and compared to a matrix of genetic elements in an attempt to deduce which GMOs are present in a sample. This can be followed by event specific screening to confirm the possible presence of an unintentional or unauthorized LMO.

In an effort to work towards establishing a baseline of the existing approaches that are currently used, we invite participants to share their experiences on their challenges and progress in the detection of unauthorized and unintentionally released LMOs. Below are some guiding questions to initiate the discussion:

1. Does your regulatory context make a distinction between “unauthorized LMOs” and “unintentionally released LMOs”?
2. How does your regulatory context address the limits for potential adventitious or low level presence in its imports of LM material already authorised and being produced in other countries, but not yet approved in your country?
3. Is your laboratory capable of detecting and identifying unauthorized and/or unintentionally released LMOs both in the context of traceability (e.g. in shipments) and environmental monitoring? If so, what strategies is your laboratory employing in order to detect unauthorized and/or unintentionally released LMOs ? If not what are the barriers to adopting such a strategy?

As mentioned by participants in previous discussions, one of the challenges faced by their laboratories included the availability and access to DNA sequence information and reference materials that are needed for quality control purposes calibration of methods for the detection of LMOs and LM products and for potential research and development projects.

A reference material (RM) is defined as: “material or substance, one or more of whose property values are sufficiently homogenous and well-established to be used for calibration of an apparatus, the assessment of a measurement method, or for assigning values to materials.” ((EC) No 641/2004). Obtaining reliable RM, containing known amounts of LMO content, is important to produce reproducible accurate and precise results in the context of determining whether or not the levels of LMOs in a shipment meet national threshold requirements. One can also obtain Certified reference material (CRM) which is a “reference material, accompanied by a certificate, one or more of whose property values are certified by a procedure which establishes its traceability to an accurate realisation of the unit in which the property values are expressed, and for which each certified value is accompanied by an uncertainty at a stated level of confidence.” ((EC) No 641/2004).

Furthermore RMs serve as a tool for analytical method validation and proficiency testing, both in-house and for inter-laboratory studies. For these tests and studies to be accurate RMs must be homogenous, stable and have undergone minimal degradation, which poses challenges with their manufacture, transport and long-term storage. Additionally the harmonisation of the type of reference material that is commonly used be it a preparation of matrix based material, such as grain flour, a DNA-RM for PCR based testing or protein-RM for ELISA testing needs to be considered in order to ensure accurate quantification results, given the variability in the estimation of LMO quantity that can be obtained when using a matrix based RM as opposed to a DNA-RM or protein-RM.

Moreover, while there has been progress in the development of RMs originating from the most common commercialised LMOs, such as soy, maize or canola, reference materials are still lacking for the majority of LM species registered in the BCH, including most tropical LM crops such as papaya, cowpeas or sorghum. While some of these reference materials can be developed in-house, the availability of certified reference material for these LMOs is equally important so that laboratories can comply with their national regulations and also engage in research and monitoring activities.

Finally, another challenge related to the access to reference material concerns information-sharing and the awareness of whether a certain RM exists and how to access it. In this context an inventory of of existing reference materials could be beneficial.

Participants are invited to further discuss these issues with the challenges they face in obtaining and using RMs.

Below are some suggested points for discussion:

1. What experiences do laboratories have in accessing such materials?
2. How have laboratories been coping thus far with the challenges related to the availability and access to reference materials?
3. Would the creation of a single portal for collecting information about reference materials available worldwide be valuable?
4. How can countries with less experience in the detection of LMOs gain access to reference materials?
5. What recommendations for solutions do you propose to improve accessibility to reference materials?

Specificity, sensitivity and costs of established methods for LMO detection

Discussions will be held between 16 June - 30 June 2013
Discussions open and close at 1:00 a.m. GMT

The availability of standardised methodologies for the detection of LMOs was also a matter that was brought up by participants as a challenging issue. At each step of the analysis process, from extraction through to DNA or protein based identification methods, laboratories must consider their needs for specificity and sensitivity versus the costs to carry out their analyses.

The choice of methodologies is impacted by the levels of sensitivity and trueness required by their regulatory frameworks. For instance, methodologies that are based on protein assays as opposed to DNA assays display varying levels of sensitivity and specificity and each can potentially play a role in the testing strategy within laboratory protocols to ensure there is a thorough examination of each sample while at the same time balancing the costs associated with carrying out the tests. This may translate to the generation of workflows that rely on a broad spectrum pre-screening of samples followed by a series of event-specific methods and analysis using a GMO reference matrix to determine which LMOs are present and in what quantities.

Furthermore some regulatory frameworks have zero-tolerance regarding the presence of unauthorized LMOs in shipments and require methodologies that are robust and sensitive enough to detect low-level or adventitious presence of such LMOs. Likewise, some regulatory framework call for environmental monitoring to detect the presence of LMOs unintentionally introduced into the environment. The adventitious presence of LMOs in either shipments or environmental samples could occur at quantities that are close to a methodology’s limit of quantification (LOQ). As a result, establishing well validated and robust methodologies with well-defined LOQ and limits of detection (LOD) is important to ensure such low level presence is reliably detected and quantified.

Finally, an ever growing number of LMOs being commercialized and under development means that more and more detection methods need to be developed. This is particularly challenging when unauthorized LMOs are to be detected.

Under this discussion topic, participants are invited to discuss issues faced when choosing which methodologies to use in their laboratories, the technical issues they face and how they meet the requirements of their biosafety regulatory frameworks.

Below are some suggested points for discussion:

1. What methodologies are most commonly used in your laboratories for the various steps involved in LMO detection?
2. Are the levels of sensitivity and specificity provided by these methodologies sufficient to satisfy the needs of your National Regulatory context?
3. Where are the major cost barriers in your laboratory: research and development, accreditation costs, method validation, reagents and supplies etc.?
4. Can methods that allow for the detection of adventitious LMOs be deployed at feasible costs?

National regulatory context and current capacity for the detection of LMOs

Discussions were held between 20 May - 2 June 2013
Discussions open and close at 1:00 a.m. GMT

As outlined in the concept note, one of the outcomes of the Network of Laboratories the Detection & Identification of LMOs is to develop a strategy that could help Parties in addressing their needs for capacity building in the field of detection and identification of LMOs in the context of the requirements of their national regulatory framework. This strategy will be brought for consideration by the COP-MOP at its seventh meeting as a set of recommendations from the Network.

As such, in order to establish a baseline of the current status of each Party’s capacity, participants are encouraged to share and discuss the scope of their current capabilities to detect and identify LMOs, be it through national and/or regional access to LMO detection laboratories, ease of accessibility to required laboratory materials or the feasibility of research and development projects. Furthermore participants can also elaborate on how this capacity fulfils the requirements set out by their country’s national regulatory context.

This discussion will start on 20 May and will close on 2 June 2013.

The Strategic Plan for the Cartagena Protocol on Biosafety, Operational Objective 2.3, points to Parties’ accessibility to national and/ or regional LMO detection laboratory networks that are capable of providing adequate delivery to those who use their services. The aim of this discussion is to establish an assessment of the presence and activity of currently existing networks that connect practitioners and facilitate the access of LMO detection services to a wider regional area.

Participants are invited to share their experience with networks they are members of, the scope within the network operates and how the network facilitates activities linked to LMO detection and identification.

This discussion will start on 20 May and will close on 2 June 2013.

As per a request from the COP-MOP and outlined in the concept note, one of the objectives of the Network is to compile relevant laboratory methods and current protocols that are currently in use for the detection of LMOs, in particular those that facilitate the detection of LMOs that are unauthorized or unintentionally released into the environment with the view to publish the information in a Biosafety Technical Series.

Participants are invited to upload and discuss relevant methodologies that are currently in use in their laboratories. These methodologies can range from best practices for subsampling techniques, extraction methods, use of automated sample preparation, end point PCR set up and product detection or quantitative Real-Time PCR methods amongst the various other techniques that are used in your laboratories. Documents can be attached to the posts or uploaded using the “Document Exchange” function below.

This discussion will start on 20 May and will remain open for an extended period of time.