Detection of Staphylococcus hyicus exfoliative toxin genes by dot blot hybridization and multiplex polymerase chain reaction.
We designed a novel DNA probe and novel PCR primer sets for detecting the genes coding for Staphylococcus hyicus (S. hyicus) exfoliative toxin (ET). In dot blot hybridization, the novel DNA probe hybridized with chromosomal DNA of ExhA-, ExhB-, ExhC-, ExhD-, and SHETA-producing strains. This probe also hybridized with the plasmid DNA of a SHETB-producing strain. In Southern blot hybridization, the probe hybridized with a 1.5 kb HindIII fragment of chromosomal DNA from a SHETA-producing strain. The above fragment was cloned into E. coli and the nucleotide sequence of the SHETA gene determined, this gene proved to have almost the same homology (99.6%) as the ExhB gene. It was therefore thought that SHETA is a subtype of ExhB. In multiplex PCR using five primer sets, each gene gave a band distinguishable from the others. This multiplex PCR system has high specificity among the well-known S. hyicus ET genes. Of the 69 known ET-producing S. hyicus strains, 38, 19, 10, 2 and 1 strains have exhB, exhD exhA, shetb and exhC genes, respectively.
Faculty of Veterinary Medicine, School of Veterinary Medicine, Kitasato University, Towada, Aomori, Japan.
Kenta
K
Onuma
Yusuke
Y
Uoya
Tetsuo
T
Koide
Ayumi
A
Shibata
Taishi
T
Tanabe
Hisaaki
H
Sato
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D015342:DNA Probes; D005083:Exfoliatins / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D005838:Genotype; D008969:Molecular Sequence Data; D058889:Molecular Typing; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D057788:Staphylococcus hyicus / Q000235:genetics*; D013552:Swine
D015342:DNA Probes; D005083:Exfoliatins