Detection of GM soybean by multiplex-touchdown PCR-microchip capillary electrophoresis with response surface methodology optimization.
The combination of the molecular technique, the multivariate strategy and microchip capillary electrophoresis (MCE) was applied to rapid and sensitive analysis of genetically modified (GM) soybean in food samples. A multiplex-touchdown polymerase chain reaction (PCR) system was developed for simultaneously amplifying three target sequences in Roundup Ready soybean (RRS). Response surface methodology was introduced to determine the optimal separation condition in MCE with good resolution and short analytical time. The detection of the PCR products of RRS was completed within 4 min under the optimal conditions. The specificity of the method was evaluated by testing non-GM soybean materials and three GM maize varieties (MON810, Bt176 and Bt11). A sensitivity of 0.1% GM organisms content was obtained, which was remarkably lower than the labeling threshold for transgenic food defined as 0.9% in the European regulation. The relative standard deviation of migration time was in the range of 0.17-0.95%. The proposed method was rapid, sensitive and specific and can be used to identify and detect GM soybean in food samples.
West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China.
Yongxin
Y
Li
Chinese Academy of Inspection and Quarantine Sciences, Gaobeidian North Road, Chaoyang District, Beijing 100025, People's Republic of China.
Ning
N
Su
West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China.
Bo
B
Zheng
West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China.
Jia
J
Ruan
West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China.
Yang
Y
Li
West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China.
Chunying
C
Luo
West China School of Public Health, Sichuan University, No. 17, Section 3, South Renmin Road, Chengdu, Sichuan 610041, People's Republic of China liyuanqian@hotmail.com liyuanqian46@163.com.
Yuanqian
Y
Li
D018744:DNA, Plant / Q000032:analysis / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000379:methods*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012044:Regression Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics*
D018744:DNA, Plant