Multiplex-PCR method for species identification of coagulase-positive staphylococci.
In veterinary medicine, coagulase-positive staphylococci (CoPS) other than Staphylococcus aureus have frequently been misidentified as being S. aureus strains, as they have several phenotypic traits in common. There has been no reliable method to distinguish among CoPS species in veterinary clinical laboratories. In the present study, we sequenced the thermonuclease (nuc) genes of staphylococcal species and devised a multiplex-PCR (M-PCR) method for species identification of CoPS by targeting the nuc gene locus. To evaluate sensitivity and specificity, we used this M-PCR method on 374 staphylococcal strains that had been previously identified to the species level by an hsp60 sequencing approach. We could successfully distinguish between S. aureus, S. hyicus, S. schleiferi, S. intermedius, S. pseudintermedius, and S. delphini groups A and B. The present method was both sensitive (99.8%) and specific (100%). Our M-PCR assay will allow the routine species identification of CoPS isolates from various animal species for clinical veterinary diagnosis.
Sasaki
Takashi
T
Department of Infection Control Science, Faculty of Medicine, Juntendo University, Tokyo 113-8421, Japan. hiteiha@juntendo.ac.jp
Tsubakishita
Sae
S
Tanaka
Yoshikazu
Y
Sakusabe
Arihito
A
Ohtsuka
Masayuki
M
Hirotaki
Shintaro
S
Kawakami
Tetsuji
T
Fukata
Tsuneo
T
Hiramatsu
Keiichi
K
D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D008836:Micrococcal Nuclease / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D013210:Staphylococcus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D014730:Veterinary Medicine / Q000379:methods*
D017931:DNA Primers; D004269:DNA, Bacterial; D008836:Micrococcal Nuclease