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Aspergillus collagen-like genes (acl): identification, sequence polymorphism, and assessment for PCR-based pathogen detection.

Authors
Tuntevski-K; Durney-BC; Snyder-AK; LaSala-PR; Nayak-AP; Green-BJ; Beezhold-DH; Rio-RVM; Holland-LA; Lukomski-S
Source
Appl Environ Microbiol 2013 Dec; 79(24):7882-7895
NIOSHTIC No.
20043483
Abstract
The genus Aspergillus is a burden to public health due to its ubiquitous presence in the environment, its production of allergens, and wide demographic susceptibility among cystic fibrosis, asthmatic, and immunosuppressed patients. Current methods of detection of Aspergillus colonization and infection rely on lengthy morphological characterization or nonstandardized serological assays that are restricted to identifying a fungal etiology. Collagen-like genes have been shown to exhibit species-specific conservation across the noncollagenous regions as well as strain-specific polymorphism in the collagen-like regions. Here we assess the conserved region of the Aspergillus collagen-like (acl) genes and explore the application of PCR amplicon size-based discrimination among the five most common etiologic species of the Aspergillus genus, including Aspergillus fumigatus, A. flavus, A. nidulans, A. niger, and A. terreus. Genetic polymorphism and phylogenetic analysis of the aclF1 gene were additionally examined among the available strains. Furthermore, the applicability of the PCR-based assay to identification of these five species in cultures derived from sputum and bronchoalveolar fluid from 19 clinical samples was explored. Application of capillary electrophoresis on nanogels was additionally demonstrated to improve the discrimination between Aspergillus species. Overall, this study demonstrated that Aspergillus acl genes could be used as PCR targets to discriminate between clinically relevant Aspergillus species. Future studies aim to utilize the detection of Aspergillus acl genes in PCR and microfluidic applications to determine the sensitivity and specificity for the identification of Aspergillus colonization and invasive aspergillosis in immunocompromised subjects.
Keywords
Fungi; Allergens; Respiratory-system-disorders; Pulmonary-system; Pulmonary-system-disorders; Etiology; Genes; Bioassays; Analytical-processes
CODEN
AEMIDF
Publication Date
20131201
Document Type
Journal Article
Email Address
slukomski@hsc.wvu.edu
Fiscal Year
2014
NTIS Accession No.
NTIS Price
Identifying No.
M122013
Issue of Publication
4
ISSN
0099-2240
NIOSH Division
HELD
Source Name
Applied and Environmental Microbiology
State
WV
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