生物技术通报Issue(7)：54-59,6.
添加扩增内标的沙门氏菌PCR检测方法
Development of an Internal Amplification Control in the PCR Detection for Salmonella
摘要
Abstract
By constructing an internal amplification control(IAC), this study developed a PCR system for the detection of Salmonella, which could effectively indicate false-negative results. The specific primers were designed according to the conserved gene invA in Salmonella spp.. An IAC was constructed by the compound primer technology, and finally the PCR detection system was developed. The experiment indicated that the specific 385 bp DNA fragment was amplified against 33 reference strains of Salmonella spp., while 6 strains of non-Salmonella only showed the 484 bp amplified band of IAC. The detection limit of this PCR system forpurified genomic DNA was 6.35 fg/μL. The artificial contamination assays showed that Salmonella could be detected after eight hours enrichment when the original bacterial concentration was 3.2 CFU/25 mL. A large number of food samples were also tested, and the results demonstrated that the detection system could effectively avoid the false-negatives and improve the detection accuracy.关键词
沙门氏菌/PCR检测/扩增内标/假阴性Key words
Salmonella/PCR detection/Internal Amplification/Control false-negative引用本文复制引用
杨晋,曾庆梅,张笛,刘坤,王琳,张亚军..添加扩增内标的沙门氏菌PCR检测方法[J].生物技术通报,2014,(7):54-59,6.基金项目
国家自然科学基金项目（31371844，31071556），国家高科技研究发展计划（2011AA100801），安徽省科技计划课题 （）
