论文部分内容阅读
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 for the identification of Campylobacter jejuni and Campylobacter coli. Methods 16S rRNA, hippurase and 16S-23S rRNA gene were used as target sequences to design specific primers. Multiplex PCR was used to detect 37 strains of strains. The ingene CAM nested PCR kit was used to test and verify the results. Results The multiplex PCR method could amplify the specific bands of Campylobacter jejuni and Campylobacter coli. The other control strains did not amplify the bands and had good specificity. The detection sensitivity was up to 0.81 pg / μl of jejunum Campylobacter DNA, 0.93 pg / [mu] l of Campylobacter coli DNA. The coincidence rate of multiplex PCR method and kit test results was 100%. The coincidence rate between the two methods and GB GB / T 4789.9-2008 method was over 97%. Conclusion The multiplex PCR method established in this study is quick and easy to operate, saves experimental costs, has good specificity, sensitivity and repeatability and can be used for the identification of Campylobacter.