Semighini C P, Delmas G, Park S, Amstrong D, Perlin D, Goldman G H
Faculdade de Ciências Farmacêuticas de Ribeirão Prteo, Universidade de São Paulo, Brazil.
FEMS Immunol Med Microbiol. 2001 Jul;31(1):15-9. doi: 10.1111/j.1574-695X.2001.tb01580.x.
In this study, we isolated and tested restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus based on PCR products amplified by the random amplified polymorphic DNA (RAPD) primer R108. Four DNA fragments, Afd, Af5, Af4, and Af4A, were amplified. Fragments Afd and Af5 were 85% and 88% identical at the DNA level to part of the Afut1 retrotransposon from A. fumigatus. Fragment Af4A is a duplication of fragment Af4 and both showed similarity at the amino acid level with endonucleases from other fungal retrotransposons. We used both RAPD with primer R108 and RFLP assays with Afut1, Afd, and Af4A, to determine the genetic relatedness of clinical isolates of A. fumigatus isolated sequentially from four patients colonized with A. fumigatus. The combination of these different methods suggested that the isolates infecting the four patients were not identical.
在本研究中，我们基于随机扩增多态性DNA（RAPD）引物R108扩增的PCR产物，分离并测试了烟曲霉的限制性片段长度多态性（RFLP）标记。扩增出了四个DNA片段，即Afd、Af5、Af4和Af4A。片段Afd和Af5在DNA水平上与烟曲霉的Afut1反转录转座子的一部分分别有85%和88%的同源性。片段Af4A是片段Af4的重复序列，二者在氨基酸水平上与其他真菌反转录转座子的内切核酸酶具有相似性。我们使用引物R108的RAPD方法以及针对Afut1、Afd和Af4A的RFLP分析，来确定从四名烟曲霉定植患者中依次分离出的烟曲霉临床分离株的遗传相关性。这些不同方法的联合使用表明，感染这四名患者的分离株并不相同。
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