阴道感染三种病原菌多重PCR检测方法的建立及其应用研究  

Establishment and Application of Multiplex PCR Detection Method for Three Kinds of Pathogens in Vaginal Secretions

作　　者：李娅[1] 章迪 张赟[1] 苏明权[3] 郭旭昌[1] 

机构地区：[1]西安市第四医院检验科,西安710004 [2]西安市中心血站,西安710061 [3]第四军医大学西京医院全军检验医学研究所,西安710032

出　　处：《现代检验医学杂志》2016年第2期42-45,共4页Journal of Modern Laboratory Medicine

摘　　要：目的探讨阴道感染病原体多重PCR快速检测白念珠菌、加德纳菌和阴道滴虫的方法建立及其临床应用。方法根据白念珠菌E03基因、加德纳菌ITS-23srRNA基因及阴道滴虫TVK3～TVK7基因设计特异性PCR引物，采用加热煮沸裂解法制备DNA模板，进行多重PCR扩增及琼脂糖凝胶电泳检测分析，在同一体系内完成三种阴道常见病原体的快速检测。结果经特异性试验证明所建立的方法能特异地检测出白念珠菌125bp，加德纳菌433bp和阴道滴虫300bp的目的片段，而其它非目的菌株均不能扩增出相应片段；敏感性试验证实白念珠菌为10cfu／ml，加德纳菌为10cfu／ml，阴道滴虫为20cfu／ml；对30份已知临床标本和10份模拟混合标本进行检测均可检测出相应的病原体。结论实验证明该研究所建立的白念珠菌、加德纳菌和阴道滴虫的多重PCR体系，具有灵敏度高、特异度强，可一步同时检测阴道分泌物中三种病原体，用于阴道感染的早期快速诊断。Objective To establish a multiplex PCR method for rapid detection of Candida albicans,Gardiner bacteria and Trichomonas vaginalis. Methods The specific PCR primers were designed on the basis of Candida albicans EO3 gene,ITS- 23srRNA gene of Trichomonas vaginalis,Gardiner bacteria and TVK3-TVK7 gene. The DNA template were preparated by the heating boiling lysis method. The specific gene of Candida albicans,Gardiner bacteria and Trichomonas vaginalis were detected by the method of PCR amplification and agarose gel electrophoresis. Multiple PCR amplification and agarose gel electrophoresis were used to detect the three kinds of commonpathogens in the same system. Results The objective gene fragment of Candida albicans 125 bp,Gardiner 433 bp and Trichomonas vaginalis 300 bp were detected rapidly and specific- ally by the multiplex PCR method. Specific experiments showed that the proposed method could specifically detect Calbicans 125 bp,Gardnerella 433 bp and Trichomonas vaginalis 300 bp fragment,and othernon target strains are not. The sensitivity experiments confirmed that Candida albicans 10 cfu/ml,Gardiner bacteria 10 cfu/ml, Trichomonas vaginalis 20 cfu/ml. The corresponding pathogens were detected in 30 clinical specimens and 10 simulated mixed specimens. Conclusion A sensitive, specific one-step detection for Candida albicans ,Gardiner bacteria and Trichomonas vaginalis multi-PCR system has been initially established,by which three kinds of pathogens in vaginalsecretions can be diagnosed early and rapidly.

关 键 词：白念珠菌 加德纳菌 阴道滴虫 多重PCR 

分 类 号：R372[医药卫生—病原生物学] Q503[医药卫生—基础医学]