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Phänotypische und genotypische Charakterisierung von Referenzstämmen der Gattung Aspergillus
Author(s) -
Rath P.M.
Publication year - 2001
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1046/j.1439-0507.2001.00620.x
Subject(s) - biology , rapd , aspergillus flavus , microbiology and biotechnology , aspergillus fumigatus , aspergillus nidulans , fungi imperfecti , gel electrophoresis , genotype , aspergillus niger , polyacrylamide gel electrophoresis , genetics , biochemistry , gene , enzyme , genetic diversity , population , demography , sociology , mutant
Summary. Twenty‐five culture collection strains from four Aspergillus species ( A. fumigatus n = 8, A. flavus n = 8, A. niger n = 4, A. nidulans n = 5) were characterized by four methods: (i) determination of patterns in an assimilation assay; (ii) protein pattern of whole mycelial cell lysates in the sodium dodecyl sulphate (SDS)‐polyacrylamide gel electrophoresis (PAGE); (iii) reactivity of a pool serum obtained from cystic fibrosis patients with mycelial lysates in the immunoblot; and (iv) random amplification of polymorphic DNA (RAPD) with eight primers having arbitrary or repetitive sequences. In the assimilation assay the A. fumigatus strains showed identical patterns in contrast to the strains of the species A. flavus , A. niger , and A. nidulans , which each showed four patterns. In the SDS‐PAGE no differences in the band patterns in the A. fumigatus strains were found, in contrast to the A. flavus (three patterns), A. nidulans (five patterns) and A. niger strains (two patterns). The immunoblot patterns were characteristic for each species with bands at 62 and 17/18 kDa in the A. fumigatus strains, at 51 and 18 kDa in the A. flavus strains, at 51 kDa in the A. niger strains, and at 51, 40 and 17/18 kDa in the A. nidulans strains allowing, however, no intraspecies typing. In the RAPD assay four out of eight primers gave interpretable patterns with 3–20 bands. None of the primers showed sufficient discriminatory power when used alone. However, when combining the results of two of the primers (5′‐GTA TTG CCC T‐3′ and 5′‐GAT AGA TAG ATA GAT A‐3′) all strains except two A. fumigatus strains could be clearly separated from each other. It is concluded that the the RAPD assay showed the most discriminatory power in all Aspergillus species investigated. In contrast to the phenotypically similar A. fumigatus strains, the strains of the species A. flavus , A. nidulans and A. niger differed in their phenotypic characteristics. The presented data of strains from international culture collections may serve as basis for interlaboratory standardization of typing methods.