拓扑异构酶II基因区PCR-RFLP法鉴别常见皮肤癣菌  

Identification of common species of dermatophytes by PCR-RFLP techniques targeting topoisomerase II gene

作　　者：谭志建[1] 何甘霖[1] 丁娟[1] 李家文[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院皮肤科,武汉430022

出　　处：《中国麻风皮肤病杂志》2006年第2期114-116,共3页China Journal of Leprosy and Skin Diseases

摘　　要：目的：探讨拓扑异构酶Ⅱ基因区PCR～RFLP法鉴别常见皮肤癣菌的可行性。方法：用真菌拓扑异构酶Ⅱ基因区通用引物dPsD1、dPsD2，先后对6种34株临床常见皮肤癣菌和2株白念珠菌以及4株曲霉的DNA进行巢式PCR（nested PER）扩增，扩增后的阳性产物分别用限制性内切酶HincⅡ和HinfⅠ酶切，进行限制性片段长度多态性（RFLP）分析，根据结果来鉴定皮肤癣菌并在种的水平上区分这6种常见皮肤癣菌。结果：通用引物dPsD1对6种皮肤癣菌均能扩增出3390bp、dPsD2能扩增出2380bp的片段，而白念珠菌和曲霉则未见阳性扩增条带。扩增出的产物分别经HincⅡ和HinfⅠ酶切后表现为58—1670bp长短不等的片段组合，根据这些特异性条带谱可以区分这6种临床常见皮肤癣菌。结论：拓扑异构酶Ⅱ基因区的巢式PCR—RFLP方法，是鉴定和区分常见皮肤癣菌的有效方法。Objective： To determine the feasibility of identification of common species of dermatophytes by PCR - RFLP techniques targeting topoisomerse Ⅱ gene. Methods： The DNA topoisomerase Ⅱ gene of six common dermatophytes, Candida albicans and Aspergilli were amplified by consensus primer dPsD1, followed by a second PCR with primers dPsD2. The products generated by dPsD2 were digested with restriction enzyme Hinc Ⅱ and Hinf Ⅰ separately. Results： A DNA fragment of about 3 390 bp was amplified by using consensus primer dPsD1 from the genomic DNA of each dermatophyte species. The products of dPsD2 was 2 380 bp and the restriction profiles of Hinc Ⅱ and Hinf Ⅰ were 58bp and 1 670 bp. No positive products could be seen in the amphcation of Candida albicans and Aspergilli. With the method of PCR - RFLP, all of the six dermatophytoses could be identified at species level. Condusion： The PCR- RFLP identification targeting the DNA topoisomerase Ⅱ gene is rapid and efficient, and can be used as a tool for the identification of the common dermatophyte species clinically.

关 键 词：皮肤癣菌 拓扑异构酶 巢式PCR RFLP Ⅱ基因区 PCR—RFLP法 

分 类 号：R756[医药卫生—皮肤病学与性病学]