交叉引物等温扩增技术检测沙门氏菌

摘要

To establish a new rapid screening method for detection of Salmonella-cross-primer amplification. On the basis of sequence analysis of the Salmonella, specific primers and probe were designed, we developed cross-primer isothermal amplification method. The specificity of the method was evaluated by 19 strains of Salmonella and 35 negative strains of Enterobacteriaceae.The sensitivity of the method was evaluated by testing the quantitative DNA,pure bacteria counted. The method established in this article was highly specific and sensitive. The limit of decetion was 101 fg/test for the genomic DNA, 102 cfu/mL for pure bacterial culture. The method was highly sensitive,specific, easy to operate and does not require any complex equipment. It is very useful for detection of Salmonella in the port or underdeveloped areas.%　　建立一种新型沙门氏菌快速筛选检测方法--交叉引物恒温扩增检测方法.针对沙门氏菌设计特异性引物及探针,建立交叉引物等温扩增法进行检测.用19株沙门氏菌,35株近源菌进行特异性试验；通过定量DNA、纯菌液计数检测进行灵敏度验证.建立方法具有很好的特异性；增菌液检测灵敏度为102 cfu/mL,DNA检测灵敏度为101 fg/test.建立的交叉引物恒温检测方法特异性好、灵敏度高,不需要复杂仪器,对口岸快速筛查、基层或偏远经济不发达地区顺利开展检测具有实际意义.