高分辨率溶解曲线在4种皮肤癣菌快速鉴定中的初步应用

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英文篇名：Clinical application of high resolution melting curve analysis for rapid identification of dermatophytes 作者：刘红芳 ; 吴宜泉 ; 刘应辉 ; 谢振谋 ; 陈阳霞 ; 黄进梅 ; 曾维英 英文作者：LIU Hong-fang;WU Yi-quan;LIU Ying-hui;XIE Zhen-mou;CHEN Yang-xia;HUANG Jin-mei;ZENG Wei-ying;Dermatology Hospital,Southern Medical University; 关键词：皮肤癣菌 ; HRM ; rDNA ; ITS 英文关键词：dermatophyte;;HRM;;rDNA ITS 中文刊名：LPFZ 英文刊名：Journal of Diagnosis and Therapy on Dermato-Venereology 机构：南方医科大学皮肤病医院; 出版日期：2018-06-28 出版单位：皮肤性病诊疗学杂志 年：2018 期：v.25;No.120 基金：国家自然科学基金青年项目(编号:81601746);; 广东省公益研究与能力建设专项基金(编号:2014A020212679) 语种：中文; 页：LPFZ201803001 页数：5 CN：03 ISSN：44-1671/R 分类号：7-11

摘要

目的:建立准确、快速鉴定红色毛癣菌、指(趾)间毛癣菌、犬小孢子菌、堇色毛癣菌等4种常见皮肤癣菌的高分辨率溶解曲线(HRM)的方法。方法:根据目的基因序列及参考文献合成基因扩增引物,应用4HRM方法鉴定4种常见皮肤癣菌,并利用rDNA ITS测序验证其准确性。结果:4种皮肤癣菌表现出不同的HRM曲线及Tm值,后续rDNA ITS测序证实了HRM溶解曲线鉴别不同种皮肤癣菌的准确性。结论:本研究采用PCR-HRM分析方法,获得4个常见皮肤癣菌的不同溶解曲线,可用于临床该类皮肤癣菌感染的初步鉴定。
        Objective:To develop a rapid and accurate identification method for four strains of dermatophytes [Trichophyton rubrum(T.rubrum),Microsporum canis(M.canis),Trichophyton interdigitale(T.interdigitale),Trichophyton violaceum(T.violaceum) ] using high-resolution melting(HRM) analysis.Methods:According to the gene sequences,primers were synthesized.Four strains of dermatophytes were identified by using HRM analysis,followed by verification of accuracy with r DNA ITS sequencing.Results:HRM analysis showed that four strains of dermatophytes displayed different melting curves and Tm values.The accuracy of HRM analysis was validated by r DNA ITS sequencing.Conclusion:HRM analysis method can accurately identify dermatophyte strains,and can be used to preliminarily identify fungal infections in clinics.

引文

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