1. The Advisory Committee for Biotechnology and 21st Century Agriculture (AC21) Accuracy for Quantitative Testing of Biotechnology Derived Grains Steven N. Tanner, Director, Technical Services Division December 5, 2003

2. ▪ Facilitate U.S. grain markets ▪ When testing is needed, ensure reliability and credibility of testing for transgenic grains and oil seeds GIPSA and Biotechnology

3. USDA/GIPSA Proficiency Program ▪ Improve the reliability of testing for biotechnology events ▪ 86 organizations now participating ▪ Corn events: T25, CBH351, MON810, GA21, E176, Bt11 and NK603; TC1507 and MON863 to be added in FY 2004 ▪ Soybeans: Glyphosate tolerance

4. USDA/GIPSA Proficiency Program Global Program AreaFeb 2002Nov. 2003 Africa 0 1 Australia 0 1 Asia 1 8 Europe 5 39 South America 1 10 North America 15 27 Total Participants22 86

5. USDA/GIPSA Proficiency Program ▪ Voluntary participation ▪ Participant anonymity ▪ Offered quarterly ▪ Qualitative and quantitative ▪ Results on GIPSA’s biotechnology web page

6. General Observations ▪ Participants use protein and DNA-based tests ▪ Qualitative results correct > 90 % of the time Quantitative Sample Results: ▪ Average results were lower than target level ▪ Coefficients of variation (CV’s) ranged from ~30 to >100 % ▪ False negatives/positives were < 10 % except at lowest levels (i.e., 0.1 %) USDA/GIPSA Proficiency Program Observations

7. Relative Measurement Variability Analyte STD*CV (%)* 1 ppb 0.45 45 1 ppm 0.16 16 1 % 0.04 4 DNA-Based Testing** STD Range CV Range 0.1 % 0.06-0.50 80-190 1.5 % 0.50-1.15 42-93 5.0 % 1.12-4.20 36-90 *Horwitz predicted values **Actual values reported in Proficiency Program (% by wt)

8. ▪ Methods - Patent Protection/Intellectual Property - Limited official protein-based methods - No globally recognized methods for conventional or real time PCR ▪ Reference Materials - Limited availability - No general agreement (seed, grain, plasmids, etc.) Testing Challenges

9.  International Collaboration of Quantitative Methods  Real Time PCR Techniques  DNA Isolation Procedures  DNA Quantitation Technologies ▪ International Reference Materials: GIPSA and NIST Workshop on June 27, 2003 Addressing the Challenges

10. ▪ Protein and DNA reference materials ▪ Matrix reference materials ▪ Global convention for expressing results GIPSA/NIST Workshop Recommendations

11. ▪ USDA GIPSA ▪ NIST ▪ Life Science Organizations ▪ Institute for Reference Materials and Measurements (IRMM/Europe) ▪ National Research Council (NRC/Canada) ▪ National Food Research Institute (NFRI/Japan) ▪ International Organizations (Codex, ISO, CEN, AOAC-I, AACC, and AOCS) Key Organizational Cooperation

12. USDA Biotechnology Webpage www.usda.gov/gipsa/ biotech/biotech.htm