Bart-Delabesse E, Humbert J F, Delabesse E, Bretagne S
Laboratoire de Parasitologie-Mycologie, Hôpital Henri Mondor, Créteil, France.
J Clin Microbiol. 1998 Sep;36(9):2413-8. doi: 10.1128/JCM.36.9.2413-2418.1998.
The use of microsatellites as highly polymorphic DNA markers for the typing of isolates of Aspergillus fumigatus was investigated. Four CA repeats were selected by screening an A. fumigatus DNA library with a (CA)10 oligonucleotide. Primers flanking these CA repeats were designed to amplify each locus. One primer of each pair was labeled with a fluorophore, and the PCR products were analyzed with an automatic sequencer and the GeneScan software. For each primer set and for a given isolate, one band was detected and was assigned to an allele because A. fumigatus is haploid. With 50 clinical isolates, 50 environmental isolates, and 2 reference strains we obtained 12, 11, 10, and 23 different alleles for the four CA microsatellites, respectively (discriminatory power, 0.994). The results were identical by whatever DNA extraction technique was used. Interestingly, no clustering between environmental and clinical isolates was observed, suggesting that every isolate is potentially pathogenic. Microsatellite markers appear suitable for use in large epidemiological studies of invasive aspergillosis.
研究了使用微卫星作为高度多态性DNA标记对烟曲霉分离株进行分型的情况。通过用(CA)10寡核苷酸筛选烟曲霉DNA文库，选择了四个CA重复序列。设计了位于这些CA重复序列两侧的引物以扩增每个位点。每对引物中的一个用荧光团标记，PCR产物用自动测序仪和GeneScan软件进行分析。对于每个引物组和给定的分离株，检测到一条带并将其指定为一个等位基因，因为烟曲霉是单倍体。对于50株临床分离株、50株环境分离株和2个参考菌株，我们分别在四个CA微卫星上获得了12、11、10和23个不同的等位基因(鉴别力，0.994)。无论使用何种DNA提取技术，结果都是相同的。有趣的是，未观察到环境分离株和临床分离株之间的聚类现象，这表明每个分离株都有潜在的致病性。微卫星标记似乎适用于侵袭性曲霉病的大型流行病学研究。
