摘要
目的建立鉴定空肠弯曲菌和结肠弯曲菌的多重PCR(mPCR)方法。方法分别以16S rRNA、马尿酸酶和16S-23S rRNA基因为靶序列设计特异性引物,建立多重PCR方法检测37株菌株样品,同时采用ingene CAM nested PCR检测试剂盒检测验证,进行结果比较分析。结果该多重PCR方法可扩增出空肠弯曲菌和结肠弯曲菌的特异性条带,其他对照菌株均未扩增出条带,具有较好的特异性;检测敏感性可达0.81 pg/μl空肠弯曲菌DNA,0.93 pg/μl结肠弯曲菌DNA。多重PCR方法和试剂盒检测结果的符合率为100%,二者与国标GB/T 4789.9—2008方法的符合率达97%以上。结论本试验建立的多重PCR方法操作快速方便、节约试验成本,具有较好的特异性、敏感性和重复性,可用于弯曲菌的鉴定。
Objective To establish a multiplex PCR （mPCR） method to identity Campylobacterjejuni and Campylobacter coli. Methods Specific primer pairs were designed based on the sequence of 16S rRNA gene, hippuricase gene and 16S- 23S rRNA gene. 37 strains were detected by the mPCR and gene CAM nested PCR assay kit. Results The results showed that the species-specific product could be detected after amplification of the DNA template of C. jejuni and C. coli, while other strains could not be detected. The sensibility for detection of C. jejuni and C. coli was 0. 81 and 0.93 pg/l, respectively. The coincidence rate mPCR method and nested PCR assay kit was 100%. Coincidence of the two methods with the national standard method were also over 97%. Conclusion This new method was rapid, convenient, highly specific, sensitive and repeatable. It could be used for rapid identification of Campylobacter spp..
出处
《中国食品卫生杂志》
北大核心
2014年第2期119-123,共5页
Chinese Journal of Food Hygiene
基金
农业部"引进国际先进农业科学技术"重点项目-948项目(2011-G14(2))
