A rapid screening method for Panton-Valentine leucocidin-positive community-acquired methicillin-resistant Staphylococcus aureus belonging to multilocus sequence type 30 and its related clone using a combination of multiplex PCR and pulsed-field gel electrophoresis.
Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA), which is often positive for Panton-Valentine leucocidin (PVL), is increasingly noted as an emerging pathogen worldwide. In Japan, PVL-positive CA-MRSA belonging to multilocus sequence type (ST) 30 has spread and caused, for example, pediatric death due to community-acquired pneumonia and severe pelvic abscesses in an athlete. In this study, we investigated a new rapid screening method for PVL-positive ST30 CA-MRSA and its related clone by a combination of multiplex polymerase chain reaction (M-PCR) and pulsed-field gel electrophoresis (PFGE). For M-PCR, the targets of the assay were the five genes for PVL, collagen adhesin, bone sialoprotein adhesin, methicillin resistance, and S. aureus-specific thermostable nuclease. Only PVL-positive ST30 CA-MRSA strains produced all five bands in M-PCR. With PFGE, Japanese strains and most foreign strains of PVL-positive ST30 CA-MRSA shared the same pattern. Moreover, PFGE distinguished current PVL-positive CA-MRSA ST30/spa19 strains from previous PVL-positive MRSA ST30/spa43 strains (which were isolated at the time of nosocomial MRSA outbreaks in the late 1980s and early 1990s) in Japan. Thus, the M-PCR assay rapidly, and the M-PCR/PFGE combination assay more precisely, discriminated between PVL-positive ST30 CA-MRSA (or its related clone) and PVL-positive CA-MRSA belonging to other ST types such as ST1, 8, 59, and 80, PVL-negative CA-MRSA, hospital-acquired MRSA, methicillin-susceptible S. aureus, or coagulase-negative staphylococci (CNS), including MRCNS. This screening method is more useful than genotyping for routine work in many clinical laboratories.
Reva
Ivan
I
Division of Bacteriology, Department of Infectious Disease Control and International Medicine, Niigata University Graduate School of Medical and Dental Sciences, 757 Ichibanchou, Asahimachidori, Niigata, 951-8510, Japan.
Higuchi
Wataru
W
Takano
Tomomi
T
Singur
Olga
O
Ozaki
Kyoko
K
Isobe
Hirokazu
H
Yabe
Shizuka
S
Saito
Kohei
K
Baranovich
Tatiana
T
Enany
Symaa
S
Otsuka
Taketo
T
Potapov
Vladimir
V
Nishiyama
Akihito
A
Yamamoto
Tatsuo
T
D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics*; D017714:Community-Acquired Infections / Q000382:microbiology*; D003428:Cross Infection / Q000382:microbiology; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D005098:Exotoxins / Q000235:genetics*; D006801:Humans; D007564:Japan; D007956:Leukocidins / Q000235:genetics*; D055624:Methicillin-Resistant Staphylococcus aureus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013203:Staphylococcal Infections / Q000382:microbiology*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D005098:Exotoxins; D007956:Leukocidins; C078284:Panton-Valentine leukocidin