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Past Discussions 2013

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Opening of Discussion: Emerging Techniques for LMO Detection [#5421]
Dear Participants of the Laboratory Network,

Welcome to the third round of discussion of the Network of Laboratories for LMO Detection & Identification. The topic that will be focused on in this forum for the next two weeks is regarding “Emerging Techniques for LMO Detection”.

In the page of the discussion (http://bch.cbd.int/onlineconferences/portal_detection/discussions.shtml#theme5), we have provided a brief introduction on the topic and potential techniques and research activities that may be underway in your laboratories. 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. 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?

We are looking forward to a fruitful discussion!
Best Regards,
Dina
posted on 2013-07-14 22:53 UTC by Dina Abdelhakim, SCBD
This is a reply to 5421 RE: Opening of Discussion: Emerging Techniques for LMO Detection [#5425]
PCR technology is adopted by the Biosafety Unit of the Royal Scientific Society (RSS) of Jordan, the Unit is performing routine tests for detection of GM products (food and feed) and conducting training courses. Currently, the RSS Biosafety Unit is adopting JRC-GMO lab technologies for detection of GM products; and we are satisfied with these (JRC-GMO lab) technologies. However we have some work on identification of genetic elements which are not supposed to be present in the authorized GM crops. Such work requires DNA sequencing and bioinformatic skills. Good experience in bioinformatic is required to conduct alignment analysis of new DNA sequences in authorized GM products.
posted on 2013-07-17 13:06 UTC by Dr. Nisreen AL-Hmoud, Royal Scientific Society
This is a reply to 5421 RE: Opening of Discussion: Emerging Techniques for LMO Detection [#5428]
Dear all,

thank you for opening the new round of discussion focused on the emerging techniques for LMO detection.  This is a very welcomed topic. There are more then 60 methods validated and used in the EU network of LMO detection laboratories. This valorouse experience would be extremily useful to be sared with other countries in the region, including the EE countries. I would like to mention a number of trainings and events organised by TAEX, in collaboration with EFSA for CEE laboratory people with the topic of LMO detection.

At the same time it would be extremily useful to extend the regional collaboration in detection methodologies, laboratory supplies and materials, as well as increasing communication and exchange of information between laboratories. To be mentioned, some communication is maintained between the EE laboratories by individual contacts, happens time to time, have some limitations in technical posibilities, as well as in the methodologies used. The scientific simposiums and conferences, as well as trainings and workshops organised with regional participation can facilitate this process and contributing to better communication on the topic of scientific methods of detection, as well as increasing capacities for development of regulations for validation of methods.

The PCR quantitative and qualitative DNA detection is usually used for LMOs detection. The protocols in national language are available. It is requested by countries authorities to develop express metodologies for LMO detection with the sutisfied level of precision to be useful during the custom control, and also monitoring of agro-and food products inclusive in the local market.  We would be interested to learn from other participants to the forum about their experience and new methods used as classical laboratory detection and also as experess methods available and useful.

Best wishes,
Angela Lozan
ME, Moldova
posted on 2013-07-18 00:26 UTC by Ms. Angela Lozan, Republic of Moldova
This is a reply to 5428 RE: Opening of Discussion: Emerging Techniques for LMO Detection [#5437]
Dear all,

Thanks to the Secretariat for opening this discussion. I would like to share some of our experiences regarding emerging techniques for LMO detection.

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?

Yes, we are developing qRT-PCR based on “MGB – Minor Groove Binder” chemistry.  MGB probes contain a non-fluorescent quencher at the 3´ end and a minor groove binder at the 3´ end. The minor groove binder increases the melting temperature (Tm) of probes, allowing the use of shorter probes. MGB probes exhibit greater differences in Tm values between matched and mismatched probes, which provide more accurate discrimination. MGB probes are patented by Applied Biosystems but as far as I am aware, their patents are close to expire.

References: Nucleic Acids Res. 2000 January 15; 28(2): 655–661.
Mol Cell Probes. 2006 Oct; 20(5):311-6.

2. What techniques are laboratories researching to improve upon their existing protocols?

Commercial ELISA tests. We have bought Quantitative Elisa kits for Cry1ab detection in maize but so far, no success has been achieved to produce a reliable standard curve with the material supplied by the company. Therefore, we are currently testing three different Elisa kits with our own home-made standard curve, using purified protein. I would like to ask participants if they have standardized home-made Elisa protocols and/or reliable commercial kits currently available.

Reference:  Food Agr Immunol. 2012 June; 23(2):  99-121.

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?

We try to follow the criteria adopted by the JRC and ISO guidelines.

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?

To our understanding, the lack of purified plant proteins is one of the major technical barrier to all protein-based detection methods.  Consequently, the lack of such plant versions has an impact in the production of antibodies against these proteins. All commercial available antibodies are against bacterial versions of these proteins. If we consider the case of MON810, there is documented possibility of the production of recombinant mRNAs derived from Cry1ab and genes present at the junction region, thus producing Cry1ab protein variants. These variants and possible post-translation modifications of these proteins might not be detected by current detection methodologies that rely on antibodies produced from bacterial protein versions. Greater impact of such barrier can be observed in safety studies. We are now focusing our efforts in producing large amounts of plant proteins versions using cell tissue culture techniques to enhance transgene expression but we are still in the very beginning of this process.

Another barrier is the detection of new-types of LMOs, such as RNAi-based LMOs or LMOs that produces double-stranded RNA. We have currently published our concerns regarding the safety of such organisms; which includes, of course, concerns about its detection. To our uderstanding, there is little information/guidelines about dsRNA detection methodologies regarding LMOs.

Reference: Environ Int. 2013 May; 55:43-55.


Best regards,

Sarah Agapito.
posted on 2013-07-22 19:02 UTC by Ms. Sarah Agapito-Tenfen, Brazil