食品科学 ›› 2010, Vol. 31 ›› Issue (22): 403-406.doi: 10.7506/spkx1002-6630-201022089

• 分析检测 • 上一篇    下一篇

双重PCR 方法检测沙门氏菌和空肠弯曲菌

陈 诺1,唐善虎1,*,陈进会2,岑璐伽1,李 雪1,龙 虎1   

  1. 1.西南民族大学生命科学与技术学院
    2. 东莞出入境检验检疫局
  • 收稿日期:2010-07-07 修回日期:2010-10-21 出版日期:2010-11-25 发布日期:2010-12-29
  • 通讯作者: 唐善虎 E-mail:stang01@126.com
  • 基金资助:

    西南民族大学研究基金项目

Development of a Duplex PCR for Detection of Salmonella spp and Campylobacter jejuni in Food

CHEN Nuo1,TANG Shan-hu1,*,CHEN Jin-hui2,CEN Lu-jia1,LI Xue1,LONG Hu1   

  1. 1. College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China;
    2. Dongguan Inspection and Quarantine Bureau, Dongguan 523072, China
  • Received:2010-07-07 Revised:2010-10-21 Online:2010-11-25 Published:2010-12-29
  • Contact: TANG Shan-hu E-mail:stang01@126.com

摘要:

为建立能够同时检测食品中沙门氏菌和空肠弯曲菌的双重PCR 方法。采用沙门氏菌鞭毛基因fimY 和空肠弯曲菌马尿酸酶基因hipO 设计特异性引物,并对影响PCR 扩增的主要因素——引物浓度、退火温度、Mg2+ 浓度因素进行优化,比较单一PCR 和双重PCR 的检测效果。结果表明:采用单一PCR 法检测沙门氏菌和空肠弯曲菌时,灵敏度分别可达到3.98pg 和4.05pg;而采用双重PCR 检测时,灵敏度较单一PCR 法有所下降,沙门氏菌和空肠弯曲菌检出限量分别为398pg 和40.5pg。本研究建立的特异性强和灵敏度高的双重PCR 检测方法,可为实现食品中沙门氏菌和空肠弯曲菌的同时检测提供新方法。

关键词: 沙门氏菌, 空肠弯曲菌, 双重PCR, 检测

Abstract:

A rapid assay for simultaneous detection of Salmonella spp and Campylobacter jejuni in food was developed. The primers were designed based on the fimY gene of Salmonella spp and hipO gene of Campylobacter jejuni. Annealing temperatures, Mg2+ concentrations, and primer concentrations were optimized. The detection limit was 3.98 pg for Salmonella spp and 4.05 pg for Campylobacter jejuni using the single PCR method. However, the detection limit of the duplex PCR method was significantly higher, which was 398 pg, 40.5 pg for Salmonella spp and Campylobacter jejuni, respectively. In this study, a duplex PCR with high specificity and sensitivity was developed and it would provide useful information for the simultaneous detection of Salmonella spp and Campylobacter jejuni in food.

Key words: Salmonella spp, Campylobacter jejuni, Duplex PCR, Detection

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