摘要
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。
Objective To develop a multiplex PCR method to detect pathogenic fungi of onychomycosis. Methods The 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. Results The 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 10^2cfu/ml. Conclusion Multiplex PCR might be a rapid,sensitive and specific method for the diagnosis of onychomycosis.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2007年第1期10-13,共4页
The Chinese Journal of Dermatovenereology
关键词
聚合酶链反应
甲真菌病
诊断
Polymerase chain reaction
Onychomycosis
Diagnosis
