应用多重PCR快速检测空肠弯曲菌  

Rapid detection of Campylobacter jejuni by multiplex PCR technique

作　　者：吴家林[1] 沙丹[1] 马广源[1] 季亚勇[1] 孙燕萍[1] 张敬平[1] 

机构地区：[1]江苏省无锡市疾病预防控制中心,江苏无锡214023

出　　处：《中国卫生检验杂志》2011年第11期2685-2687,共3页Chinese Journal of Health Laboratory Technology

基　　金：卫生部科研基金(WKJ2006-2-10);无锡市卫生系统"十一五"重大综合发展项目(WZD0601);无锡市卫生系统科研项目(XM0901)

摘　　要：目的:建立能快速、特异检测空肠弯曲菌的多重PCR技术。方法:选用针对空肠弯曲菌外膜蛋白A(mapA)基因和马尿酸酶(hipO)基因的2对引物,在同一扩增体系中进行PCR,优化反应体系,测定特异性和灵敏度,并进行了鸡肉模拟样品检测。结果:该方法扩增目的基因片段分别为589 bp和323 bp,特异性和灵敏度均高。细菌纯培养物的检测灵敏度为105 cfu/ml,鸡肉模拟样品42℃预增菌36 h后检测灵敏度能达到101 cfu/ml。结论:初步建立能快速、灵敏、特异地测定空肠弯曲菌的多重PCR检测技术。Objective：To establish a multiplex PCR technique for rapid and specific detection of Campylobacter jejuni.Methods： Two sets of primers from mapA and hipO gene of Campylobacter jejuni were selected and added into one amplification system to perform PCR.The system was optimized,the specificity and sensitivity of this system were evaluated,and artificially-contaminated chicken were detected.Results： The assay was designed to amplify the 589 bp and 323 bp regions of corresponding genes mapA and hipO with high sensitivity and specificity.Sensitivity of the assay was 105 cfu/ml for bacteria samples and 101 cfu/ml for Campylobacter jejuni in artificially-contaminated chicken with incubation at 42℃ for 36 hours.Conclusion： A rapid,specific and sensitive multiplex PCR technique for the simultaneous detection of Campylobacter jejuni has been studied primarily.

关 键 词：多重PCR 空肠弯曲菌 mapA基因 hipO基因 

分 类 号：R446.5[医药卫生—诊断学]