Huang Xin, Zhai Congcong, You Qimin, Chen Hongjun
Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China,
Anal Bioanal Chem. 2014 Jul;406(17):4243-9. doi: 10.1007/s00216-014-7791-y. Epub 2014 Apr 16.
The requirement to monitor the presence of genetically modified organisms (GMO) in a variety of marked products has generated an increasing demand for reliable, rapid, and time and cost-effective analytical methods. Here we report an on-site method for rapid detection of cauliflower mosaic virus promoter (CaMV 35S), a common element present in most GMO, using cross-priming amplification (CPA) technology. Detection was achieved using a DNA-based contamination-proof strip biosensor. The limit of detection was 30 copies for the pBI121 plasmid containing the CaMV 35S gene. The certified reference sample of GM maize line MON810 was detectable even at the low relative mass concentration of 0.05%. The developed CPA method had high specificity for the CaMV 35S gene, as compared with other GM lines not containing this gene and non-GM products. The method was further validated using nine real-world samples, and the results were confirmed by real-time PCR analysis. Because of its simplicity, rapidity, and high sensitivity, this method of detecting the CaMV 35S gene has great commercial prospects for rapid GMO screening of high-consumption food and agriculture products.
监测各种标记产品中转基因生物（GMO）的存在的需求，使得对可靠、快速且具有时间和成本效益的分析方法的需求不断增加。在此，我们报告一种使用交叉引物扩增（CPA）技术快速检测花椰菜花叶病毒启动子（CaMV 35S）的现场方法，CaMV 35S是大多数转基因生物中存在的常见元件。使用基于DNA的防污染条带生物传感器进行检测。对于含有CaMV 35S基因的pBI121质粒，检测限为30个拷贝。转基因玉米品系MON810的认证参考样品即使在相对质量浓度低至0.05%时也可检测到。与其他不含该基因的转基因品系和非转基因产品相比，所开发的CPA方法对CaMV 35S基因具有高度特异性。该方法使用9个实际样品进一步验证，结果通过实时PCR分析得到证实。由于其简单、快速和高灵敏度，这种检测CaMV 35S基因的方法在对高消费食品和农产品进行转基因生物快速筛选方面具有巨大的商业前景。
