title: Rapid and accurate identification of human-associated staphylococci by use of multiplex PCR.
abstract: Although staphylococci are identified by phenotypic analysis in many clinical laboratories, these results are often incorrect because of phenotypic variation. Genetic analysis is necessary for definitive species identification. In the present study, we developed a simple multiplex-PCR (M-PCR) for species identification of human-associated staphylococci, which were as follows: Staphylococcus aureus, S. capitis, S. caprae, S. epidermidis, S. haemolyticus, S. hominis, S. lugdunensis, S. saprophyticus, and S. warneri. This method was designed on the basis of nucleotide sequences of the thermonuclease (nuc) genes that were universally conserved in staphylococci except the S. sciuri group and showed moderate sequence diversity. In order to validate this assay, 361 staphylococcal strains were studied, which had been identified at the species levels by sequence analysis of the hsp60 genes. In consequence, M-PCR demonstrated a sensitivity of 100% and a specificity of 100%. By virtue of simplicity and accuracy, this method will be useful in clinical research.
authors.affiliation: Department of Infection Control Science, Faculty of Medicine, Juntendo University, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Hirotaki
authors.forename: Takashi
authors.initials: T
authors.lastname: Sasaki
authors.forename: Kyoko
authors.initials: K
authors.lastname: Kuwahara-Arai
authors.forename: Keiichi
authors.initials: K
authors.lastname: Hiramatsu
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008836:Micrococcal Nuclease / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000382:microbiology; D013210:Staphylococcus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial; D008836:Micrococcal Nuclease