title: Comparison of phenotypic and genotypic methods for the species identification of coagulase-negative staphylococcal isolates from bovine intramammary infections.
abstract: Coagulase-negative staphylococci (CNS) are the most frequently isolated pathogens from cows with intramammary infection (IMI). Although API STAPH ID 20, a commercially available identification system, and PCR-restriction fragment length polymorphism (PCR-RFLP) of the gap gene (gap PCR-RFLP) have been successfully applied for the identification of CNS isolates from human specimens, their accuracy in the identification of veterinary isolates has not been fully established. In this study, we identified 263 CNS isolates from bovine IMI at species level by partial 16S rRNA gene sequence analysis as the definitive test. Species identification obtained using partial 16S rRNA gene sequence analysis was compared to results from the API STAPH ID 20 and gap PCR-RFLP analysis. Eleven different CNS species were identified by partial 16S rRNA gene sequence analysis. Only 76.0% (200/263) of the species identification results obtained by API STAPH ID 20 matched those obtained by partial 16S rRNA gene sequence analysis, whereas 97.0% (255/263) of the species identification results obtained by the gap PCR-RFLP analysis matched those obtained by partial 16S rRNA gene sequence analysis. The gap PCR-RFLP analysis could be a useful and reliable alternative method for the species identification of CNS isolates from bovine IMI and appears to be a more accurate method of species identification than the API STAPH ID 20 system.
authors.affiliation: Department of Veterinary Clinical Science, Washington State University, Pullman, WA 99164-6610, USA.
authors.forename: Joo Youn
authors.initials: JY
authors.lastname: Park
authors.forename: Lawrence K
authors.initials: LK
authors.lastname: Fox
authors.forename: Keun Seok
authors.initials: KS
authors.lastname: Seo
authors.forename: Mark A
authors.initials: MA
authors.lastname: McGuire
authors.forename: Yong Ho
authors.initials: YH
authors.lastname: Park
authors.forename: Fred R
authors.initials: FR
authors.lastname: Rurangirwa
authors.forename: William M
authors.initials: WM
authors.lastname: Sischo
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Bohach
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002417:Cattle; D003030:Coagulase / Q000378:metabolism; D005260:Female; D020459:Genes, rRNA; D005838:Genotype; D008414:Mastitis, Bovine / Q000382:microbiology*; D008828:Microbiological Techniques / Q000379:methods / Q000662:veterinary*; D010641:Phenotype; D012150:Polymorphism, Restriction Fragment Length; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013045:Species Specificity; D013203:Staphylococcal Infections / Q000382:microbiology / Q000662:veterinary*; D013210:Staphylococcus / Q000145:classification* / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D003030:Coagulase; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S