中国动物检疫2018，Vol.35Issue(3)：81-85,5.DOI:10.3969/j.issn.1005-944X.2018.03.022
志贺氏菌检测和分群PCR方法的建立
Study on PCR Method for Shigella Detection and Group Differentiation
摘要
Abstract
In order to improve the current methods for molecular biological detection of Shigella,primers were designed respectively according to the genes of invC,rfc,wbgZ and rfpB of Shigella,and a PCR method was established to identify Shigella,S.flexneri,S.sonnei and S.dysenteriae.At the same time,taking the primers designed from ompA gene as a reference,the method was verified by specific and artificial inoculation tests.The results showed that specific reaction were observed in both 17 Shigella strains and 19 non-Shigella strains.When the enrichment broth was detected directly by PCR,the detection limit of Shigella in instant food was 1~3 cfu/(25 g·mL-1),including pasteurized milk,ice cream,acidophilus milk,cheese,cooked meat and sau sage,and the detection limits of Shigella in raw milk,raw meat and powdered milk were ≤ 12,27 and ≤ 27 cfu/(25 g·mL-1),respectively. When PCR was done after isolation and culture of suspected bacteria,the detection limit of all samples was 1-3 cfu/(25 g·mL-1),which was consistent with the traditional culture method. In conclusion,the PCR method was rapid, sensitive and specific,and it was suitable for routine analysis of Shigella.关键词
志贺氏菌/检测和分群/PCRKey words
Shigella/detection and differentiation/PCR分类
农业科技引用本文复制引用
王海艳,赵治国,催强,郭文秀,陈宇飞,赵林立..志贺氏菌检测和分群PCR方法的建立[J].中国动物检疫,2018,35(3):81-85,5.基金项目
国家质检总局科技计划项目(2014IK095) （2014IK095）
