Gene sequences and specific detection for Panton-Valentine leukocidin.
A new category of methicillin-resistant Staphylococcus aureus (MRSA), called community-acquired MRSA (CA-MRSA), has emerged worldwide. In contrast to previous MRSA, most CA-MRSA carries the Panton-Valentine leukocidin (PVL) genes (lukPVSF) as a virulence genetic trait. Sequence analysis of the lukPVSF gene of a Japanese isolate demonstrated that the gene has more similarity to methicillin-susceptible S. aureus from France than MRSA from the United States. Based on the sequences, we developed a real-time PCR assay for the three key genes of CA-MRSA; that is, lukPVSF, mecA (for methicillin resistance), and spa (for S. aureus). Dual or triple assay for lukPVSF, mecA, and spa in one test tube became possible. The detection limit of the assay with probe and SYBR Green methods was between 2.7 and 2.7 x 10(1) CFU/ml. The assay detected PVL-positive MRSA in clinical (blood) isolates.
Nakagawa
Saori
S
Division of Bacteriology, Department of Infectious Disease Control and International Medicine, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Taneike
Ikue
I
Mimura
Daigo
D
Iwakura
Nobuhiro
N
Nakayama
Teruko
T
Emura
Tomoko
T
Kitatsuji
Maki
M
Fujimoto
Atsushi
A
Yamamoto
Tatsuo
T
D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins; D002648:Child; D002874:Chromosome Mapping / Q000379:methods*; D017714:Community-Acquired Infections / Q000235:genetics; D005098:Exotoxins; D016680:Genome, Bacterial; D006801:Humans; D007956:Leukocidins / Q000235:genetics*; D016106:Methicillin Resistance / Q000235:genetics; D015336:Molecular Probe Techniques; D046915:Penicillin-Binding Proteins; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013203:Staphylococcal Infections / Q000235:genetics*; D013205:Staphylococcal Protein A / Q000235:genetics; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D005098:Exotoxins; D007956:Leukocidins; C078284:Panton-Valentine leukocidin; D046915:Penicillin-Binding Proteins; D013205:Staphylococcal Protein A; C487789:mecA protein, Staphylococcus aureus