狐阴道加德纳氏菌PCR诊断方法的建立与应用  

Establishment and application of Gardnerella vaginalis of fox by PCR

作　　者：颜赟[1] 刘悦竹 吴召强 李宏梅[1] 孙杰[1] 程艳丽[1] 刁有祥[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]山东立鼎生物技术研究所,山东济南250022

出　　处：《西北农林科技大学学报（自然科学版）》2010年第5期32-35,共4页Journal of Northwest A&F University(Natural Science Edition)

基　　金：山东省中青年科学家奖励基金项目(2008BS07002)

摘　　要：【目的】建立快速检测狐阴道加德纳氏菌的PCR诊断方法。【方法】根据阴道加德纳氏菌16S rRNA基因序列设计1对特异性引物,以分离的狐阴道加德纳氏菌菌体DNA为模板进行PCR扩增,建立狐阴道加德纳氏菌的PCR检测方法,并对该方法的特异性、敏感性及其可行性进行检验。【结果】特异性试验结果表明,狐阴道加德纳氏菌能扩增出437 bp的特异性核酸片段,而大肠杆菌、沙门氏菌、绿脓杆菌、金黄色葡萄球菌的扩增结果均为阴性。敏感性试验结果表明,PCR的最低检出限量为500 mL-1。利用建立的PCR检测方法,对9株从山东省不同地区分离的疑似狐阴道加德纳氏菌菌株进行检测,结果有4株为阳性。【结论】研究建立的狐阴道加德纳氏菌PCR快速诊断方法,敏感性高、特异性强,可1次检测多个样品。【Objective】 PCR diagnostic method of rapid detection of Gardnerella vaginalis of fox was established.【Method】 On the basis of the Gardnerella vaginalis 16S rRNA gene,one pair of primers were designed.It took the separated Gardnerella vaginalis of fox DNA as a template and went on PCR amplification.So PCR detection method of Gardnerella vaginalis of fox was established,and its with specificity and sensitivity were studied.【Result】 It showed by the specific test results that Gardnerella vaginalis of fox can amplify 437 bp specific DNA fragment.However,the amplification results of E.coli,Salmonella pullorum,Bacillus pyocyaneus,Staphylococcus were amplified negatively.The result of the sensitiveities test showed that at least 500 mL-1 Gardnerella vaginalis of fox was detected with the approach.With the approach of the established PCR the 9 suspected strains isolated from different regions of Shandong Province were detected and 4 strains were positive.【Conclusion】 The PCR diagnostic method of rapid detection of Gardnerella vaginalis of fox is successfully established,and its sensitive and can detect more samples at are time.

关 键 词：狐 狐阴道加德纳氏菌 PCR 诊断方法 

分 类 号：S865.23[农业科学—野生动物驯养] S858.92[农业科学—畜牧兽医]