大豆粉转基因成分能力验证样品的检测分析

目的 提高实验室对转基因大豆定性检测的准确性和检测人员专业技术水平, 增强实验室竞争力。方法 通过核酸蛋白仪器分析法和单重实时荧光PCR (simplex real-time fluorescence PCR)法对样品内源基因扩增的循环阈值的影响来比较2种方法, 分析2种DNA提取试剂盒的提取质量。对标准SN/T 1204-2016《植物及其加工产品中转基因成分实时荧光PCR定性检验方法》扩增反应体系中DNA模板量进行优化后, 对样品进行检测。再依据标准SN/T 1202-2010《食品中转基因植物成分定性PCR检测方法》的要求, 对样品进行检测。最后对2个标准的检测结果进行比对。结果 通过内源基因扩增循环阈值的数据确认, 更能准确反映试剂盒提取的DNA是否满足后续外源基因的分析检测要求。2种标准方法对19-N578和19-M913 2个待测样品的检测显示3种外源基因CaMV35S、NOS、CP4-EPSPS均为阴性, 19-N578和19-M913待测样品均为非转基因大豆。结论 本次能力验证获得满意评价。DNA提取质量的评估, 体系中DNA模板量的优化, 检测方法的选择和实验的质量控制都是影响能力验证结果的重要因素。

Detection and analysis of genetically modified proficiency testing samples of soybeans flour

Objective To improve the accuracy of results of qualitative detection ability of genetically modified in soybeans flour and the professional technical level of the detection personnel, and enhance the competitiveness of the laboratory. Methods The DNA extraction quality of 2 DNA extraction kits was analyzed by nucleic acid protein instrument analysis and simplex real-time fluorescence PCR method by comparing the cycle threshold for endogenous gene amplification in samples. The amount of DNA template in PCR amplification system of standard SN/T 1204-2016 Protocal of the real-time PCR method for detecting genetically modified plants and their derived products was optimized, and then the samples were tested. The samples were also detected according SN/T 1202-2010 Protocal of the qualitative polymerase chain reaction for detecting genetically modified plant components in food. The results of the two standards were compared. Results The confirmation of data for detection of the amplification cycle threshold of endogenous genes could more accurately reflect that whether the DNA extracted from the kit met the requirements of subsequent analysis of detection of foreign genes. The foreign genes including CaMV35S, NOS and CP4-EPSPS were all negative in both of the two samples 19-N578 and 19-M913 by the two methods. The two samples were non-transgenic soybeans. Conclusion The ability verification results are satisfied. The evaluation of DNA extraction quality, the optimization of DNA template amount, the selection of detection methods, and the quality control of experiments are the important factors to influence the proficiency testing results.