Aspergillus Collagen-Like Genes (acl): Identification, Sequence Polymorphism, and Assessment for PCR-Based Pathogen Detection

Author:

Tuntevski Kiril,Durney Brandon C.,Snyder Anna K.,LaSala P. Rocco,Nayak Ajay P.,Green Brett J.,Beezhold Donald H.,Rio Rita V. M.,Holland Lisa A.,Lukomski Slawomir

Abstract

ABSTRACTThe genusAspergillusis 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 ofAspergilluscolonization 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 theAspergilluscollagen-like (acl) genes and explore the application of PCR amplicon size-based discrimination among the five most common etiologic species of theAspergillusgenus, includingAspergillus fumigatus,A. flavus,A. nidulans,A. niger, andA. terreus. Genetic polymorphism and phylogenetic analysis of theaclF1gene 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 betweenAspergillusspecies. Overall, this study demonstrated thatAspergillusaclgenes could be used as PCR targets to discriminate between clinically relevantAspergillusspecies. Future studies aim to utilize the detection ofAspergillusaclgenes in PCR and microfluidic applications to determine the sensitivity and specificity for the identification ofAspergilluscolonization and invasive aspergillosis in immunocompromised subjects.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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