title: Aspergillus collagen-like genes (acl): identification, sequence polymorphism, and assessment for PCR-based pathogen detection.
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.
authors.affiliation: Department of Microbiology, Immunology, and Cell Biology.
authors.forename: Kiril
authors.initials: K
authors.lastname: Tuntevski
authors.forename: Brandon C
authors.initials: BC
authors.lastname: Durney
authors.forename: Anna K
authors.initials: AK
authors.lastname: Snyder
authors.forename: P Rocco
authors.initials: PR
authors.lastname: Lasala
authors.forename: Ajay P
authors.initials: AP
authors.lastname: Nayak
authors.forename: Brett J
authors.initials: BJ
authors.lastname: Green
authors.forename: Donald H
authors.initials: DH
authors.lastname: Beezhold
authors.forename: Rita V M
authors.initials: RV
authors.lastname: Rio
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Holland
authors.forename: Slawomir
authors.initials: S
authors.lastname: Lukomski
mesh_terms: D001228:Aspergillosis / Q000175:diagnosis* / Q000382:microbiology; D001230:Aspergillus / Q000235:genetics / Q000302:isolation & purification*; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D016000:Cluster Analysis; D003094:Collagen / Q000235:genetics*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D009172:Mycology / Q000379:methods*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D013183:Sputum / Q000382:microbiology
chemical_list: D004271:DNA, Fungal; D005656:Fungal Proteins; D003094:Collagen