The detection of T-Nos,a genetic element present in GMOs,by cross-priming isothermal amplification with real-time fluorescence |
| |
Authors: | Fang Zhang Liu Wang Kai Fan Jian Wu Yibin Ying |
| |
Institution: | 1. College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China 2. Key Laboratory of Equipment and Informatization in Environment Controlled Agriculture, Ministry of Agriculture, Hangzhou, China
|
| |
Abstract: | An isothermal cross-priming amplification (CPA) assay for Agrobacterium tumefaciens nopaline synthase terminator (T-Nos) was established and investigated in this work. A set of six specific primers, recognizing eight distinct regions on the T-Nos sequence, was designed. The CPA assay was performed at a constant temperature, 63 °C, and detected by real-time fluorescence. The results indicated that real-time fluorescent CPA had high specificity, and the limit of detection was 1.06?×?103 copies of rice genomic DNA, which could be detected in 40 min. Comparison of real-time fluorescent CPA and conventional polymerase chain reaction (PCR) was also performed. Results revealed that real-time fluorescent CPA had a comparable sensitivity to conventional real-time PCR and had taken a shorter time. In addition, different contents of genetically modified (GM)-contaminated rice seed powder samples were detected for practical application. The result showed real-time fluorescent CPA could detect 0.5 % GM-contaminated samples at least, and the whole reaction could be finished in 35 min. Real-time fluorescent CPA is sensitive enough to monitor labeling systems and provides an attractive method for the detection of GMO. Figure ? |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|