Establishment of National Genetically Modified Product Detection Laboratories and Detection Method Complex

1. Establishment of National Genetically Modified Product Detection Laboratories and Detection Method Complex ZHU Shui-fang, Ph.D GMOs detection center Institute of Animal and Plant Quarantine

2. Research contents 1. Detection technology of GMOs Real-time PCR DNA Microarray Captured PCR-ELISA Quicktest strip 2. Publications for GMOs detection 12 SN standard protocols 8 GB standard protocols 4 for regulation and detection technology 3. Construction of laboratories for GMOs detection

3. Detection technology of GMOs • Real-time PCR • DNA Microarray • Captured PCR-ELISA • Quicktest strip

4. Real-time PCR Screening Kits Target genes: CaMV 35S Promotor, Nos teminator , NptII, Bar, FMV promotor, Pat Reference gene: 18S rRNA Quantitative Kits Roundup Ready soybean Bt176 Maize Event-specific detection kits GTS40-3-2, Bt176, Mon810, Bt11, GA21, T25, RT73

5. Real time PCR - TaqMan™ Technology

6. Positive sample Negative sample Ct value

7. Screening Detection of GM potato and rapeseed

8. Amplification of EPSPS gene Amplification of lectin gene Quantitative detection of Roundup Ready Soybean

9. Real-time PCR kits

10. DNA Microarray Total: 46 genes 31 inserted genes e.g. EPSPS 7 reference genes e.g. lectin 8 event-specific genes e.g. Roundup Ready Soybean Genes involved in microarray research

11. DNA Microarray Technology

12. DNA Microarray Screening chip Event-specific chip Event-specific sequence Versatile chip 9 genes Species chip Comprehensive chip Bt176, Bt11, Mon810, T14/25, GA21, CBH-351 maize Bioscien toamto Mon531 cotton GM Soybean, maize rapeseed, cotton P35s, Pnos, NptII CaMV35S! Nos, FMV35S Bar, Pat1(maize) Pat2 (rapeseed) Detection of food and feed

13. Specific detection for RUR soybean Multiple PCR and hybridization results of rapeseed Multiple PCR and hybridization results of GM maize Specific detection for Mon810 maize

14. Specific detection for Bt11 maize Multiple PCR and hybridization results of soybean Specific detection for GA21 maize Specific detection for Bt176 maize Multiple PCR and hybridization results of cotton

15. DNA Microarray kits

16. Captured PCR-ELISA Capture of target nucleic acid molecule PCR-ELISA

17. Adding sample Target DNA molecule impurity molecule washing Affinity capture of target DNA molecule

18. PCR product probe electrophoresis hybridization PCR-ELISA Procedure

19. control test negative positive Quicktest strip for Cry1Ab/Ac

20. Publications for GMOs detection 4 for regulation and detection technology 12 SN standard protocols 8 GB standard protocols

21. GMOs regulation and detection technology 2 for regulation, 2 for detection technology SN and GB protocols for GMOs detection 12 SN (issued in 2003), 8 GB (will soon be issued) Detection methods (revising, ISO 7th Working group) Bioscientomato, Mon531 cotton, RT73 rapeseed, Edible Oil

22. Publications of GMOs regulation and detection technology

23. 12 SN standard protocols •The general requirements for the laboratories for gene detection and identification • Protocol of the Qualitative Polymerase Chain Reaction (PCR) for Detecting Genetically Modified Component in Soybeans • Protocol of the Polymerase Chain Reaction for Detecting Genetically Modified Components in Potatoes • Protocol of the Polymerase Chain Reaction for Detecting Genetically Modified Components in cotton • Protocol of the real-time PCR for detecting genetically modified plants and their derived products • Protocol of PCR for detection of genetically modified feed • Protocol of the Polymerase Chain Reaction for Detecting Genetically Modified Components in maize • Methods of sampling and preparation of samples for detection of genetically modified components in plants and their derived products • Protocol of the Detection Methods for Genetically Modified tobacco • Protocol of the polymerase chain reaction for detecting genetically modified in rapeseeds • Protocol of the Qualitative Polymerase Chain Reaction for Detecting Genetically Modified Plant Components in Food • Protocol of the Qualitative Polymerase Chain Reaction for Detecting Genetically Modified Plant Components in Edible Oil

24. 8 GB standard protocols • Detection of genetically modified organisms and derived products - Nucleic acid extraction • Detection of genetically modified organisms and derived products –Gene chip detection • Detection of genetically modified organisms and derived products –Protein based methods •Detection of genetically modified organisms and derived products - Methods for sampling and sample preparation • Detection of genetically modified organisms and derived products - Quantitative nucleic acid based methods • Detection of genetically modified organisms and derived products - Qualitative nucleic acid based methods •Detection of genetically modified organisms and derived products - General requirements for laboratories • Detection of genetically modified organisms and derived products - General requirements and definitions

25. Construction of labs for GMOs detection • 25 labs for GMOs detection • Proficiency test • Training of detection technology

26. National Genetically Modified Product Detection Laboratories and Detection Method Complex Labs for GMOs detection Detection methods Information Exchanging and training of detection technology Construction of labs Requirements for detection labs Proficiency test of detection labs Methods for GMOs detection GMO websites Kits for GMOs detection GMO databases Issue of standard protocols for GMOs detection DNA detection methods Protein detection methods

27. Thank you!