PCR-RFLP法直接检测分枝杆菌皮肤感染的临床应用

Clinical applicability of PCR-RFLP in detection of mycobacterium infections of skin

目的:评价分枝杆菌热休克蛋白65(Hsp65)基因检测分枝杆菌皮肤感染标本的敏感性和特异性。方法:多聚酶链反应-限制性片段长度多态性(PCR-砒LP)的10%非变性聚丙烯酰胺凝胶法直接检测Hsp65基因,PCR-RFLP检测阳性的标本再经Hsp65基因和16S rRNA DNA基因测序验证。结果:与培养法相比,PCR-RFLP法检测临床分枝杆菌皮肤感染的敏感性为30%(3/10),特异性为100%(10/10)。结论:分枝杆菌PCR-RFLP的10%非变性聚丙烯酰胺凝胶法可用于分枝杆菌皮肤感染的临床检测。

Objective： To assess the sensitivity and specificity of polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）based on hsp65 by 10% non-denaturizing polyacrylamide gel in the detection of mycobacterium infections of skin. Methods： Twenty clinical skin specimens suspected of mycobacterium infections were detected and identified by culture and PCR- RFLP directly and confirmed by sequencing of hsp65 and 16S rRNA DNA genes. Results： Compared with culture method, the sensitivity and the specificity of PCR- RFLP were 30% （3/10） and 100% （10/10）. Conclusion： PCR- RFLP based on hsp65 with 10% polyacrylamide gel can be clinically applied in the detection of mycobacterium infections of skin.