基于等位基因特异性PCR原理建立鸡白痢沙门氏菌PCR方法

摘要

Based on the nuclei polymorphism of rfbS gene sequence of Salmonella pullorum at site 237 and 598 compared with S.gallinarum,a pair of allele-specific primers were designed and synthesized,and an allele-specific PCR method for detecting S.pullorum was developed consistent with result of the serological and biochemical method and the sensitivity of the PCR assay was 18 pg DNA and 4.7× 104 CFU/mL cultural liquid.The results showed that the developed allele-specific PCR was a very rapid,sensitive and specific molecular tool for the detection and identification of S.pullorum isolates.%根据鸡白痢沙门氏菌与鸡伤寒沙门氏菌的rfbS基因在第237和598位碱基的不同,设计和合成等位基因特异性PCR引物,建立快速检测鸡白痢沙门氏菌的PCR方法,并应用该法对鸡白痢沙门氏菌临床分离样品进行了PCR鉴定.结果显示,该PCR方法能特异性地鉴定鸡白痢沙门氏菌,检测灵敏度达18pg/μLDNA,4.7×104CFU/mL菌液,表明建立的等位基因特异性PCR方法能准确而快速地鉴定鸡白痢沙门氏菌.