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目的应用基于28S rRNA基因序列的巢式PCR的方法快速诊断甲真菌病。方法液氮冷冻甲标本,微量法提取DNA,应用特异性引物,巢式PCR方法扩增DNA。结果本组共收集180例直接镜检和培养均为阳性的甲标本,First PCR 144例阳性,Nest PCR 168例阳性。其中127例标本经PCR扩增可产生约137bp目的片段,为红色毛癣菌;26例标本经PCR扩增可产生约102bp目的片段,为须癣毛癣菌;10例标本经PCR扩增可产生约445bp目的片段,为白念珠菌;5例标本经PCR扩增可产生约170bp目的片段,为曲霉。巢式PCR敏感性为93.33%(168/180),特异性为100%(168/168)。结论此方法是一种快速、敏感、特异的诊断甲真菌病的方法。
Objective To rapidly diagnose onychomycosis using the nested PCR method based on the 28S rRNA gene sequence. Methods The samples were cryopreserved in liquid nitrogen and DNA was extracted by micro-method. The specific primers and nested PCR were used to amplify DNA. Results A total of 180 A specimens were collected directly from both microscopy and culture. First PCR was positive in 144 cases and Nest PCR was positive in 168 cases. Among them, 127 samples were amplified by PCR to produce a fragment of about 137 bp, which was Trichophyton rubrum; 26 samples amplified about 102 bp by PCR, which was Trichophyton mentagrophytes; 10 samples were amplified by PCR Resulting in about 445bp fragment of interest, Candida albicans; 5 specimens by PCR amplification can produce about 170bp fragment of interest, as Aspergillus. The sensitivity of nested PCR was 93.33% (168/180) and the specificity was 100% (168/168). Conclusion This method is a rapid, sensitive and specific method for the diagnosis of onychomycosis.