title: Molecular genotyping of Acinetobacter spp. isolated in Arizona, USA, using multilocus PCR and mass spectrometry.
abstract: Acinetobacter spp. are a diverse group of Gram-negative bacteria frequently implicated in nosocomial infections. Genotypic methods have been instrumental in studying Acinetobacter, but few offer high resolution, rapid turnaround time, technical ease and high inter-laboratory reproducibility, which has hampered understanding of disease incidence, transmission patterns and diversity within this genus. Here, we further evaluated multilocus PCR electrospray ionization/mass spectrometry (PCR/ESI-MS), a method that is simple and robust, and provides both species characterization and strain-level resolution of Acinetobacter spp. on a single platform. We examined 125 Acinetobacter isolates from 21 hospitals, laboratories and medical centres spanning four counties in Arizona, USA, using PCR/ESI-MS. We compared PCR/ESI-MS with an in-house amplified fragment length polymorphism (AFLP) genotyping scheme. PCR/ESI-MS demonstrated that Acinetobacter spp. from Arizonan hospitals had similar species and strain distributions to other US civilian hospitals. Furthermore, we showed that the PCR/ESI-MS and AFLP genotypes were highly congruent, with the former having the advantages of robust inter-laboratory reproducibility, rapid turnaround time and simple experimental set-up and data analysis. PCR/ESI-MS is an effective and high-throughput platform for strain typing of Acinetobacter baumannii and for identification of other Acinetobacter spp., including the emerging nosocomial pathogens Acinetobacter pittii and Acinetobacter nosocomialis.
authors.affiliation: Menzies School of Health Research, Darwin, NT 0810, Australia.|Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: Derek S
authors.initials: DS
authors.lastname: Sarovich
authors.affiliation: Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Colman
authors.affiliation: Menzies School of Health Research, Darwin, NT 0810, Australia.|Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: Erin P
authors.initials: EP
authors.lastname: Price
authors.affiliation: IBIS Biosciences, Carlsbad, CA 92008, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Massire
authors.affiliation: Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: Alex T
authors.initials: AT
authors.lastname: Von Schulze
authors.affiliation: Arizona Department of Health Services, Phoenix, AZ 85007, USA.
authors.forename: Victor
authors.initials: V
authors.lastname: Waddell
authors.affiliation: Arizona Department of Health Services, Phoenix, AZ 85007, USA.
authors.forename: Shoana M
authors.initials: SM
authors.lastname: Anderson
authors.affiliation: IBIS Biosciences, Carlsbad, CA 92008, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Ecker
authors.affiliation: Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Liguori
authors.affiliation: Translational Genomics Research Institute, Flagstaff, AZ 86001, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Engelthaler
authors.affiliation: IBIS Biosciences, Carlsbad, CA 92008, USA.
authors.forename: Rangarajan
authors.initials: R
authors.lastname: Sampath
authors.affiliation: Translational Genomics Research Institute, Flagstaff, AZ 86001, USA.|Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Keim
authors.affiliation: IBIS Biosciences, Carlsbad, CA 92008, USA.
authors.forename: Mark W
authors.initials: MW
authors.lastname: Eshoo
authors.affiliation: Northern Arizona University, Center for Microbial Genetics and Genomics, Flagstaff, AZ 86011, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Wagner
mesh_terms: D000150:Acinetobacter / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D054458:Amplified Fragment Length Polymorphism Analysis; D001130:Arizona; D015373:Bacterial Typing Techniques / Q000379:methods; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D058885:Multilocus Sequence Typing / Q000379:methods*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S