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

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 Research contents

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

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

Real time PCR - TaqMan™ Technology

Positive sample Negative sample Ct value

Screening Detection of GM potato and rapeseed

Amplification of EPSPS geneAmplification of lectin gene Quantitative detection of Roundup Ready Soybean

Real-time PCR kits

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

DNA Microarray Technology

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

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

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

DNA Microarray kits

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

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

electrophoresishybridization probe PCR product PCR-ELISA Procedure

Quicktest strip for Cry1Ab/Ac control test positive negative

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

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 7 th Working group ) Bioscien tomato, Mon531 cotton, RT73 rapeseed, Edible Oil

Publications of GMOs regulation and detection technology

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

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

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

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

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