关键词: 转基因大豆, 重组酶聚合酶扩增技术, 分子检测, PCR

Abstract: The goal of this work is to develop a fast and easy-operating RPA detection method for the GM soybean to monitor the illegal planting and spreading of the GM soybeans in China. Five pairs of RPA primers were initially designed based on the conserved sequences of CaMV-35S promoter and the NOS terminator in GM soybeans. PCR reactions were further performed to screen the primers and a set of RPA primers with the high sensitivity and specificity,Nos163,was selected for constructing RPA test strips. Reaction temperatures and times of RPA were also optimized. No transgenic component was detected from bean sprouts and fresh green soybean in market,as well as soy samples planted in field in Nanjing area using our RPA detection technique. All evidences suggest that RPA detection method is more sensitive than PCR,and can be performed under normal temperature without depending on the expensive equipment. The test results can be easily visualized,therefore which makes it an effective approach for the rapid detection of GM soybean.

Key words: transgenic soybean, recombinase polymerase amplification, molecular detection, PCR