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Past Discussions 2013

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Opening of the Discussion: Specificity, sensitivity and costs of established methods for LMO detection [#5391]
Dear Participants of the Laboratory Network,

Welcome to the second round of discussion of the Network of Laboratories for LMO Detection & Identification. The topic that will be focused on for the next two weeks is regarding “Specificity, sensitivity and costs of established methods for LMO detection”.

In the page of the discussion, we have provided a brief overview of the importance of developing and adopting well-validated methodologies to carry out the detection assays, and highlighted some considerations that could have an impact on the selection of appropriate methodologies. We have also listed a few guiding questions (below) to help initiate the discussion. However please feel free to bring up other related issues that you would like to discuss.

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?

We are looking forward to a fruitful discussion!
Best Regards,
The Secretariat
posted on 2013-06-16 05:09 UTC by Dina Abdelhakim, SCBD
This is a reply to 5391 RE: Opening of the Discussion: Specificity, sensitivity and costs of established methods for LMO detection [#5396]
POSTED ON BEHALF OF ANGELA LOZAN
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Dear Colleagues,

First I would like to thank the participants to the forum for sharing their experience in detection and identification of LMOs.

I would like to bring to the forum the following national data and information related the capacities and methodological aspects for detection and identification of LMOs.

The National regulatory context and current capacity for detecting LMOs:
The Biosafety law in Moldova in the art. 24 lit.c stipulates special requirements for packaging and labelling of LMOs  and products contained LMOs when placing to the market. On the label it is recommended to be written : The product contain genetically modified organisms” that aplly on the product itself and also in the accomplishing document.
The regulation prescribe that the labelling is apply when the product contain at least 1% of GMOs, and for seeds – 0,3% of the total weight of the product.

Overview of existing networks for LMO detection and identification:
I would like to mention the following existent laboratories in Moldova, Chisinau, thatare developing their capacities for LMO detection and identification.
The Laboratory of biosafety within the Faculty of Biology and Soil sciences of the State University of Moldova. The laboratory uses the DNA and proteins qualitative LMO detection. The laboratory mostly serve as scientific and training base for educational process.
The Laboratory for seed control and certification within the Agency for Food Security of the Ministry of Agriculture. A PCR Real-time machine is available for LMO detection. Laboratory is accredited for ISO17025. 
The Laboratory of molecular biology  of the University of the Academy of Sciences of Moldova is equipped with the  PCR real time machine, provided several testings and detection of Lmos in agricultural products (tobacco, tomato, corn, potato).

Compilation of laboratory methods for the detection and identification of LMOs:
The methods used at present for detection and identification are ELISA and PCR Real time.
A manual on Methodological aspects of LMO detection is published in national language and available for laboratory personnel and students: http://www.biosafety.md/public/209/ro/Aspecte_.pdf
Maria Duca, Angela Lozan, Angela Port, Aliona Glijin, Victor Lupascu. Aspecte metodologice în testarea plantelor modificate genetic. Chişinău 2008.

What methodologies are most commonly used in your laboratories for the various steps involved in LMO detection?
• The PCR qualitative and quantitative detection is used in laboratory detection;
• Primer-Screening of GM35S - CaMV and nos-terminator;
• Multiplex PCR;
• Electrophoresis amplicons;
• Analysis Southern Blot;
• Proteins expression of genes (GFP, PAT).

Are the levels of sensitivity and specificity provided by these methodologies sufficient to satisfy the needs of your National Regulatory context?
The PCR detection method is appropriate to satisfy the regulatory needs. 

Where are the major cost barriers in your laboratory: research and development, accreditation costs, method validation, reagents and supplies etc.?

The research and development, accreditation costs, method validation, reagents and supplies are the major costs that provides limitation and constrains for efficient detection and monitoring of LMOs. The existent laboratories at the moment have not been accredited at national or international level to provide services for LMOs detection. The methods have not been yet validated. There is a constrain to ensure the supply with reagents and necessary laboratory materials with regularity. The personnel is not trained in a satisfied level. There is no mechanism to provide services for detection of LMOs to people,  farmers, importers/exporters of agricultural products, seeds, food staffs etc.

Can methods that allow for the detection of adventitious LMOs be deployed at feasible costs?

The costs for detection of LMOs are usually high and not accessible for farmers, consumers, inspection services in my country. It would be helpful to have available some express methods of detection with the lower costs that could be used ex. for custom control (the small scale lots of imported agricultural products and feed), by farmers, by consumers, ONG, inspection services.

Dr.Angela Lozan, Ministry of Environment, MD
posted on 2013-06-25 13:16 UTC by Dina Abdelhakim, SCBD
This is a reply to 5396 RE: Opening of the Discussion: Specificity, sensitivity and costs of established methods for LMO detection [#5401]
Dear Secretariat and participants of the Network,

Thank you Angela for sharing your experience in Moldova. It was quite interesting to see that from my portuguese language background I could understand Moldovan quite well!

These are my first thoughts about the guiding questions proposed by the Secretariat:

1. What methodologies are most commonly used in your laboratories for the various steps involved in LMO detection?
We are currently using commercial elisa kits from various manufactures; qRT-PCR using Taqman MGB probes with self-designed primers, end-point PCR with various primers and methodologies sources, western blotting, common commercialized immunostrips and immunoprecipitation using commercial magnetic beads and commercial antibodies. We have experienced designing our own primers and probes but also, we have immunized chicken in order to produce our own antibodies. One of our main goals now is to produce large amounts of purified GM proteins from commercialized GMOs in order to produce antibodies against the plant version GM protein.

2. Are the levels of sensitivity and specificity provided by these methodologies sufficient to satisfy the needs of your National Regulatory context?
Yes.

3. Where are the major cost barriers in your laboratory: research and development, accreditation costs, method validation, reagents and supplies etc.?
I guess that reagents and supplies come first, but in general, research/development and method validation are also costly. We have never experienced accreditation costs.

4.      Can methods that allow for the detection of adventitious LMOs be deployed at feasible costs?
No.

Best wishes,

sarah
posted on 2013-06-25 18:39 UTC by Ms. Sarah Agapito-Tenfen, Brazil
This is a reply to 5391 RE: Opening of the Discussion: Specificity, sensitivity and costs of established methods for LMO detection [#5404]
POSTED ON BEHALF OF LAURA ESTHER TOVAR
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On behalf of the National Reference Center for Detection of Genetically Modified Organisms (CNRDOGM), Ministry of Agriculture of Mexico (SAGARPA)

Dear Collegues:
It is very interesting to know the techniques you use in their labs, is important to share our experiences and learn about the achievements and the challenges we face.
In the laboratory of the National Reference Center for Detection of Genetically Modified Organism (CNRDOGMS) in México, we do the 35s and TNOS detection, we can identify specific events and their quantification. We validated qPCR for detection in eleven maize events,  nine cotton events, one soybeans event and one wheat event. The challenge we face is to increase this list in our catalog of services, in quantifying modified events  we have a detection limit of 0.1%, and we have corn and soybean trials for application of dPCR allowing us to quantify the number of copies of the genetic modification.
as Dr Angela says, the cost of analysis are elevated together with aspects of staff training, accreditations costs certainly are barriers to  overcome that we share.
posted on 2013-07-01 23:01 UTC by Dina Abdelhakim, SCBD