Open AccessDiversity among clinical isolates of Histoplasma capsulatum detected by polymerase chain reaction with arbitrary primers
Author(s) -
Dangeruta Kersulyte,
Jon P. Woods,
Elizabeth J. Keath,
William E. Goldman,
Douglas E. Berg
Publication year - 1992
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.174.22.7075-7079.1992
Subject(s) - biology , polymerase chain reaction , restriction fragment length polymorphism , primer (cosmetics) , rapd , genomic dna , genetics , typing , dna profiling , dna , restriction enzyme , minisatellite , in silico pcr , genotyping , histoplasma , genotype , microbiology and biotechnology , genetic diversity , multiplex polymerase chain reaction , gene , microsatellite , histoplasma capsulatum , allele , population , histoplasmosis , chemistry , demography , organic chemistry , sociology , immunology
Clinical isolates of the fungal respiratory and systemic pathogen Histoplasma capsulatum have been placed in several different classes by using genomic restriction fragment length polymorphisms (RFLPs), but in general have not been distinguished further. We report here that a polymerase chain reaction (PCR)-based DNA fingerprinting method that has been termed arbitrary primer or random amplified polymorphic DNA (RAPD) PCR can distinguish among isolates in a single RFLP class. In this method, arbitrarily chosen oligonucleotides are used to prime DNA synthesis from genomic sites that they fortuitously match, or almost match, to generate strain-specific arrays of DNA fragments. Each of 29 isolates of RFLP class 2, the group endemic in the American Midwest, was distinguished by using just three arbitrary primers. In contrast, laboratory-derived S and E colony morphology variants of two strains were not distinguished from their R parents by using 18 such primers. Thus, the clinical isolates of H. capsulatum are quite diverse, but their genomes remain stable during laboratory culture. These outcomes suggest new possibilities for epidemiological analysis and studies of fungal populations in infected hosts.