title: Variable-Number Tandem-Repeat Analysis of Respiratory and Household Water Biofilm Isolates of "Mycobacterium avium subsp. hominissuis" with Establishment of a PCR Database.
abstract: "Mycobacterium aviumsubsp.hominissuis" is an important cause of pulmonary disease. It is acquired from environmental sources, but there is no methodology for large population studies. We evaluated the potential of variable-number tandem-repeat (VNTR) analysis. Clinical and household biofilmM. aviumisolates underwent molecular identification. Testing for IS901was done to separateM. aviumsubsp.aviumfromM. aviumsubsp.hominissuis VNTR types were defined using VNTR loci, and subtyping was performed using 3'hsp65and internal transcribed spacer (ITS) sequencing. Forty-nine VNTR types and eight subtypes ofM. aviumsubsp.hominissuis(IS901negative) were identified among 416 isolates ofM. aviumfrom 121 patients and 80 biofilm sites. Of those types, 67% were found only among patient isolates, 11% only among household water isolates, and 23% among both. Of 13 VNTR types that included ≥4 patients, the majority (61.5%) represented geographic clustering (same city). Most VNTR types with multiple patients belonged to the same 3'hsp65sequence code (sequevar). A total of 44 isolates belonging to fourM. aviumsubsp.hominissuisVNTR types (8%), including three with the rare Mav-F ITS sequence and 0/8 subspecies, produced amplicons with IS901PCR primers. By sequencing, all 44 amplicons were not IS901but ISMav6, which was recently observed in Japan but had not been previously described among U.S. isolates. VNTR analysis ofM. aviumsubsp.hominissuisisolates is easier and faster than pulsed-field gel electrophoresis. Seven VNTR loci separated 417 isolates into 49 types. No isolates ofM. aviumsubsp.aviumwere identified. The distributions of the VNTR copy numbers, the allelic diversity, and the low prevalence of ISMav6 differed from the findings for respiratory isolates reported from Japan.
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Iakhiaeva
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA.
authors.forename: Susan T
authors.initials: ST
authors.lastname: Howard
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA.
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Brown Elliott
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: McNulty
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA.
authors.forename: Kristopher L
authors.initials: KL
authors.lastname: Newman
authors.affiliation: Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA.
authors.forename: Joseph O
authors.initials: JO
authors.lastname: Falkinham
authors.affiliation: Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA.
authors.forename: Myra
authors.initials: M
authors.lastname: Williams
authors.affiliation: Pulmonary and Critical Care Division, Lankenau Medical Center, Wynnewood, Pennsylvania, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Kwait
authors.affiliation: Pulmonary and Critical Care Division, Lankenau Medical Center, Wynnewood, Pennsylvania, USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Lande
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA.
authors.forename: Ravikiran
authors.initials: R
authors.lastname: Vasireddy
authors.affiliation: Saskatchewan Disease Control Laboratory, Regina, Saskatchewan, Canada.
authors.forename: Christine
authors.initials: C
authors.lastname: Turenne
authors.affiliation: Mycobacteria/Nocardia Research Laboratory, Department of Microbiology, University of Texas Health Science Center at Tyler, Tyler, Texas, USA richard.wallace@uthct.edu.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Wallace
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D018441:Biofilms; D018834:Chaperonin 60 / Q000235:genetics; D016000:Cluster Analysis; D004251:DNA Transposable Elements; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005191:Family Characteristics; D005838:Genotype; D006801:Humans; D007564:Japan; D018598:Minisatellite Repeats; D017720:Molecular Epidemiology / Q000379:methods; D058889:Molecular Typing / Q000379:methods*; D009162:Mycobacterium avium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D058974:Phylogeography; D016133:Polymerase Chain Reaction / Q000379:methods; D014376:Tuberculosis / Q000382:microbiology*; D014871:Water Microbiology
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D004251:DNA Transposable Elements; D021903:DNA, Ribosomal Spacer; C074119:heat-shock protein 65, Mycobacterium