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红色毛癣菌、苏丹毛癣菌和古尔维利毛癣菌的聚合酶链反应鉴定

PCR identification of dermatophyte fungi Trichophyton rubrum, T. soudanense and T. gourvilii.

作者信息

Liu D, Pearce L, Lilley G, Coloe S, Baird R, Pedersen J

机构信息

*Melbourne Pathology, Collingwood, Victoria 3066 and †CSIRO Health Science and Nutrition, Parkville, Victoria 3052, Australia.

出版信息

J Med Microbiol. 2002 Feb;51(2):117-122. doi: 10.1099/0022-1317-51-2-117.

Abstract

Diagnosis of dermatophytosis employing conventional laboratory procedures has been complicated by the slow growth and varied morphological features shown by dermatophytes. After analysis of the nucleotide base sequences of a 1.2-kb fragment amplified from a dermatophyte fungus Trichophyton rubrum by arbitrarily primed PCR with random primer OPD18, a pair of primers (TRIF and TR1R) was designed and evaluated for specific identification of T. rubrum. The sensitivity of the primers TR1F and TR1R was high, as a specific PCR band of c. 600 bp was detected from as little as 7 pg of T. rubrum DNA. By examining 92 dermatophyte strains and clinical isolates, it was found that this pair of primers reacted in PCR with T. rubrum, T. soudanense and T. gourvilii through formation of the specific fragment of 600 bp, but not with any other of the dermatophyte species or varieties, fungi, yeasts or bacteria tested. As T rubrum is one of the most frequently isolated dermatophyte fungi, and T. soudanense and T. gourvilii are relatively uncommon in many parts of the world, these primers can be used for rapid, sensitive and specific identification and differentiation of T. rubrum from other fungi and micro-organisms.

摘要

采用传统实验室方法诊断皮肤癣菌病一直很复杂,因为皮肤癣菌生长缓慢且形态特征多样。通过用随机引物OPD18进行任意引物PCR从皮肤癣菌红色毛癣菌扩增出的1.2 kb片段的核苷酸碱基序列分析,设计了一对引物(TRIF和TR1R)并评估其用于红色毛癣菌的特异性鉴定。引物TR1F和TR1R的灵敏度很高,因为从低至7 pg的红色毛癣菌DNA中就能检测到约600 bp的特异性PCR条带。通过检测92株皮肤癣菌菌株和临床分离株,发现这对引物在PCR中与红色毛癣菌、苏丹毛癣菌和古氏毛癣菌反应,形成600 bp的特异性片段,但与测试的任何其他皮肤癣菌种类或变种、真菌、酵母或细菌均无反应。由于红色毛癣菌是最常分离出的皮肤癣菌真菌之一,而苏丹毛癣菌和古氏毛癣菌在世界许多地区相对不常见,这些引物可用于快速、灵敏和特异性地鉴定红色毛癣菌并将其与其他真菌和微生物区分开来。

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