高分辨熔解曲线技术检测阴道念珠菌分型的研究

Identification of Common Vaginal Candida Species by High Resolution Melting Curve Analysis

目的探讨采用高分辨熔解曲线方法鉴定常见致病性念珠菌的可行性,为外阴阴道念珠菌病的分子流行病学及病原学研究奠定基础。方法用芽管试验、厚壁孢子试验、科玛嘉(CHRO Magar)显色培养基及API20CAUX酵母菌鉴定系统对临床致病性念珠菌进行鉴定,然后对5株标准株和80株临床分离株的基因组DNA内转录间隔区进行聚合酶链反应,对扩增产物进行高分辨熔解曲线分型。结果 5株念珠菌标准株和80株临床株聚合酶链反应扩增后,通过熔解曲线技术分析产生5种不同的峰图,即得到念珠菌5种分型。结论外阴阴道念珠菌的主要致病菌是白念珠菌(77.50%),高分辨熔解曲线方法鉴定常见的致病性念珠菌特异性强、稳定性好,是传统鉴别方法的有益补充。

Objective To study the feasibility of identifing common pathogenic Candida clinically isolated from patients with vulvovaginal candidasis by high-resolution melting curve analysis （HRM） method. Methods Germ tube test, chlamydospore test, CHROMagar Candida and API20 kit system were applied to separate non-C. albicans strains from C. albicans. HRM was used to amplify internal transcribed spacer regions of ribosomal DNA from 5 type strains of 5 pathogenic Candida spp. and 80 clinical Candida isolates. Results Five types of graphic were obtainted after amplifycation from 5 Candida spp. by high-resolution melting curve a- nalysis. Conclusion Candida albicans is the most common pathogenic yeast in the vulvovaginal candidiasis （77.50%）. HRM is able to differentiate clinical Candida species and resulted in being specific, and sta- ble, which can be a good addition to traditional diagnostic approach.