Open AccessNew restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus
Author(s) -
Pizeta Semighini Camile,
Delmas Guillaume,
Park Steven,
Amstrong Donald,
Perlin David,
Goldman Gustavo Henrique
Publication year - 2001
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.2001.tb01580.x
Subject(s) - restriction fragment length polymorphism , rapd , biology , aspergillus fumigatus , primer (cosmetics) , amplified fragment length polymorphism , restriction enzyme , genetics , dna , microbiology and biotechnology , retrotransposon , genetic marker , cleaved amplified polymorphic sequence , polymerase chain reaction , genetic diversity , gene , genome , transposable element , population , chemistry , demography , organic chemistry , sociology
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.