多重PCR诊断甲真菌病的实验研究

目的初步建立多重PCR诊断甲真菌病的方法。方法选择3对引物(真菌通用引物、皮肤癣菌特异性引物和酵母特异性引物)建立三重PCR体系,采用红色毛癣菌、白念珠菌和短帚霉摸索反应条件和验证体系的适用性;并用红色毛癣菌来测定反应的灵敏度。结果筛选得到的真菌通用引物NS、皮肤癣菌特异性引物CHS1和酵母特异性引物ACT1有较好的通用性和特异性;在适宜的反应条件下,无论对单模板,还是多模板,该反应体系均能扩增出目的片段,未见明显非特异片段的干扰;该反应体系的检测灵敏度为102cfu/ml。结论多重PCR技术是一种快速、敏感和特异诊断甲真菌病的方法。

Experimental Investigation on the Detection for Pathogenic Fungi of Onychomycosis by Multiplex PCR

ObjectiveTo develop a multiplex PCR method to detect pathogenic fungi of onychomycosis.MethodsThe specificities of PCR were determined with universal fungus-specific primer,dermatophyte-specific primer and yeast-specific primer pairs respectively.Mutiplex PCR with these 3 primer pairs was used to detect 1 template (T.rubrum,C.albicans,S.brevicaulis) and 2 or 3 template mixtures.Sensitivity of multiplex PCR was measured.ResultsThe detection system was of high specificity.Multiplex PCR could amplify the corresponding 1,2 or 3 DNA fragments,depending on the number of template DNA added,and the sensitivity of multiplex PCR was 102cfu/ml.ConclusionMultiplex PCR might be a rapid,sensitive and specific method for the diagnosis of onychomycosis.