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通过聚合酶链反应(PCR)进行DNA指纹图谱分析快速鉴定念珠菌属菌种。

Rapid identification of Candida species by DNA fingerprinting with PCR.

作者信息

Thanos M, Schonian G, Meyer W, Schweynoch C, Graser Y, Mitchell T G, Presber W, Tietz H J

机构信息

Department of Dermatology, Charite Hospital, Humboldt University, Berlin, Germany.

出版信息

J Clin Microbiol. 1996 Mar;34(3):615-21. doi: 10.1128/jcm.34.3.615-621.1996.

Abstract

DNA polymorphisms in different species and strains of the genus Candida were assessed by amplifying genomic DNA with single nonspecific primers. This PCR method employed an arbitrary primer (the 10-mer AP3), a primer derived from the intergenic spacer regions (T3B), and the microsatellite primers (GTG)5 and (AC)10. Distinctive and reproducible sets of amplification products were observed for 26 different Candida and 8 other fungal species. The numbers and sizes of the amplification products were characteristic for each species. All yeast species tested could be clearly distinguished by their amplification patterns. With all primers, PCR fingerprints also displayed intraspecies variability. However, PCR profiles obtained from different strains of the same species were far more similar than those derived from different Candida species. By comparing species-specific PCR fingerprints of clinical isolates with those of reference strains, clinical isolates could be identified to the species level even if they could not be identified by routine biochemical methods.

摘要

通过使用单一非特异性引物扩增基因组DNA,评估了念珠菌属不同物种和菌株中的DNA多态性。这种PCR方法使用了一个任意引物(10聚体AP3)、一个源自基因间隔区的引物(T3B)以及微卫星引物(GTG)5和(AC)10。对于26种不同的念珠菌和8种其他真菌物种,观察到了独特且可重复的扩增产物集。扩增产物的数量和大小对每个物种而言都是特征性的。所有测试的酵母物种都可以通过其扩增模式清晰区分。使用所有引物时,PCR指纹图谱也显示出种内变异性。然而,从同一物种的不同菌株获得的PCR图谱比从不同念珠菌物种获得的图谱更为相似。通过将临床分离株的物种特异性PCR指纹图谱与参考菌株的图谱进行比较,即使临床分离株无法通过常规生化方法鉴定,也能够在物种水平上进行鉴定。

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