半巢式PCR-RFLP法快速鉴定临床标本中的皮肤癣菌  

Direct Spiecies Ldentification of Common Pathogenic Dermatophyte Fungi in Clinical Specimens

作　　者：张明莉[1] 杨国玲[1] 李文利[2] 安利佳[2] 

机构地区：[1]大连医科大学附属第一医院皮肤科,辽宁大连116011 [2]大连理工大学环境与生命学院,辽宁大连116024

出　　处：《中国皮肤性病学杂志》2007年第12期729-732,共4页The Chinese Journal of Dermatovenereology

基　　金：辽宁省科委资助项目(编号:20062154);大连市科委资助项目(编号:2003B3NS217)

摘　　要：目的探索快速鉴定皮肤癣菌病临床标本中致病菌的分子生物学方法。方法以皮肤癣菌的基因组DNA为模板,采用一对半真菌通用引物(NS5、ITS1、ITS4)行半巢式PCR扩增rDNA上的ITS段,用限制性内切酶BciT130Ⅰ及DdeⅠ酶切目的片段,凝胶电泳。结果7种65株常见皮肤癣菌培养菌株和17例临床标本的半巢式PCR-RFLP图谱特异;且临床标本与相应的培养菌株酶切图谱一致。结论半巢式PCR-RFLP方法适合培养菌株和临床标本病原菌的快速准确鉴定。Objective To develop a rapid and reliable molecular biology method to identify the common dermatophyte fungi from clinical specimens . Methods The genome DNA was extracted from cultured strains of 7 common dermatophyte fungi species and part of clinical specimen. ITS regions were amplified by semi-nested PCR（snPCR） with 3 universal primers （NS5 ,ITS1 and ITS4）for fungi. The amplifid products were digested with 2 restriction endonuclease （ BelT130 I, Dde I） RFLP. The rest of each clinical specimen was tested under microscope and cultured in Sabourand＇ s Agar medium. Then the results of RFLP compared with the culture results. Results The digestion of 7 common dermatophyte fungi produced 7 different restriction profiles. Restriction profiles of 17 clinical specimens matched respectively to them of the cultured strains ,and 14 profiles of the 17 ones were matched to culturation result completely, Conelnsion snPCR-RFLP analysis of ribosomal-DNA intergenic spacer regiongs is a valuable method of exactness and celerity for species identification of common dermatophyte fungi from clinical specimens.

关 键 词：皮肤癣菌 皮肤癣菌病 半巢式PCR RFLP 鉴定 

分 类 号：R756[医药卫生—皮肤病学与性病学]