Roque H D, Vieira R, Rato S, Luz-Martins M
Lab. Micologia, Instituto de Higiene e Medicina Tropical/CREM, Universidade Nova de Lisboa, Portugal.
J Clin Microbiol. 2006 Dec;44(12):4336-41. doi: 10.1128/JCM.00759-06. Epub 2006 Sep 27.
This report describes application of PCR fingerprinting to identify common species of dermatophytes using the microsatellite primers M13, (GACA)4, and (GTG)5. The initial PCR analysis rendered a specific DNA fragment for Microsporum audouinii, which was cloned and sequenced. Based on the sequencing data of this fragment, forward (MA_1F) and reverse (MA_1R) primers were designed and verified by PCR to establish their reliability in the diagnosis of M. audouinii. These primers produced a singular PCR band of 431 bp specific only to strains and isolates of M. audouinii, based on a global test of 182 strains/isolates belonging to 11 species of dermatophytes. These findings indicate these primers are reliable for diagnostic purposes, and we recommend their use in laboratory analysis.
本报告描述了利用微卫星引物M13、(GACA)4和(GTG)5,通过聚合酶链反应(PCR)指纹图谱技术来鉴定皮肤癣菌常见种类的应用。最初的PCR分析产生了一个针对奥杜盎小孢子菌的特定DNA片段,该片段被克隆并测序。基于该片段的测序数据,设计了正向引物(MA_1F)和反向引物(MA_1R),并通过PCR验证以确定它们在诊断奥杜盎小孢子菌中的可靠性。基于对属于11种皮肤癣菌的182个菌株/分离株的全面测试,这些引物产生了一条仅对奥杜盎小孢子菌的菌株和分离株特异的431 bp单一PCR条带。这些结果表明这些引物用于诊断目的是可靠的,我们建议在实验室分析中使用它们。