沙门氏菌PCR检测方法的建立及其在蔬菜检测中的应用

PCR detection methods of Salmonella and application in vegetables samples

为了对蔬菜中沙门氏菌进行快速检测,本论文建立了一种检测沙门氏菌的PCR方法。根据沙门氏菌特异基因设计4对引物进行PCR扩增,并通过检测不同浓度沙门氏菌菌悬液的PCR产物、测定OD600nm值与平板菌落数的对应线性关系确定沙门氏菌的检出限。结果筛选出invA基因为稳定的沙门氏菌特异性基因,此PCR技术对沙门氏菌的检出限为1550cfu/mL,对香菜的检出限为63cfu/g。通过此方法检测大连开发区180份市售蔬菜样品,6份检出沙门氏菌,检出率为3.33%,实验结果表明本文建立的沙门氏菌PCR检测方法可用于检测蔬菜中沙门氏菌的污染状况。

This study had established a PCR method of detection in order for the rapid detection of Salmonella spp. in vegetables. Four pairs of primers were designed for PCR amplification according to specific genes of Salmonella,and the detection limit of Salmonella was determined by the effects of different concentrations of Salmonella bacterial suspension of PCR product bands and the linear relation between the value of OD600nm and the colony tablet count.The invA genes were screened as stable Salmonella-specific genes, and the detection limit of the PCR technology for Salmonella was determined 1550cfu/mL and the detection limit of cilantro for Salmonella was 63cfu/g.Salmonella were detected in six of 180 vegetables samples from Dalian Development Zone by this method, and the detection rate was 3.33%.The experimental results showed that the detection method established could be used to detecte Salmonella in vegetables.