Korean J Lab Med.  2008 Aug;28(4):286-292. 10.3343/kjlm.2008.28.4.286.

Molecular Typing of Staphylococcus aureus Isolated from Blood on the Basis of Coagulase Gene Polymorphism and Toxin Genes

• Kim YK
• Kim JS
• Kim HS
• Song W
• Cho HC
• Lee KM

• KMID: 854891
• DOI: http://doi.org/10.3343/kjlm.2008.28.4.286

Abstract

BACKGROUND: Coagulase is produced by all strains of Staphylococcus aureus. The 3' coding region of the coagulase (coa) gene contains varying numbers of 81 bp tandem repeats. S. aureus produces a variety of extracellular protein toxins. Here, we typed S. aureus strains isolated from blood by coa gene restriction fragment length polymorphism (RFLP) patterns and toxin gene profiles. METHODS: A total of 120 strains of S. aureus were isolated from blood cultures during 2003-2006 at Kangdong Sacred Heart Hospital. The isolates were typed by PCR RFLP analysis of the coa gene and by multiplex PCR for detection of genes encoding enterotoxins (sea, seb, sec, sed, and see), toxic shock syndrome toxin-1 (tst), exfoliative toxins (eta and etb), mecA and femA. RESULTS: All the S. aureus strains were classified into 16 types on the basis of coa gene RFLP and could be further differentiated into 34 types according to the combined patterns of coa gene RFLP and toxin gene profiles. Of 85 methicillin-resistant S. aureus (MRSA) strains, 43 (50.6%) and 36 (42.4%) belonged to the RFLP pattern L5 and pattern L1, respectively. MRSA strains belonging to pattern L5 frequently carried tst (93.0%) or sec gene (81.4%), and strains belonging to pattern L1 frequently carried sea (88.9%) or see gene (44.4%). The rate of the pattern L5 in MRSA strains increased over the past few years and was higher in intensive care unit than in other wards. CONCLUSIONS: We typed S. aureus strains isolated from blood on the basis of coa gene RFLP and toxin genes. The strains belonging to coa gene RFLP pattern L5 and L1 appeared to be the major types of MRSA isolasted from bacteremia and revealed specific toxin gene profiles according to the coa gene RFLP patterns.

Keyword

Staphylococcus aureus; Coagulase; Restriction fragment length polymorphism; Enterotoxins; Methicillin resistance

MeSH Terms

Adolescent
Adult
Aged
Aged, 80 and over
Bacterial Proteins/genetics
Bacterial Typing Techniques
Child
Child, Preschool
Coagulase/*genetics
Female
Humans
Infant
Male
Methicillin Resistance/genetics
Middle Aged
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Staphylococcal Infections/*diagnosis/genetics/microbiology
Staphylococcus aureus/*classification/genetics/isolation & purification
Toxins, Biological/*genetics
Young Adult

Figure

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  Fig. 1. Agarose gel electrophoresis of PCR-amplified coa genes from S. aureus strains. RCR amplification with primers COA-1 and COA-2 [9] resulted in single fragment bands of four different sizes: S, M, L, and XL. Marker Lanes contain molecular size standard (100 bp ladder).

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  Fig. 2. Agarose gel electrophoresis of PCR-amplified coa genes digested with the restriction endonuclease AluI from representative strains of S. aureus. Marker Lanes contain molecular size standard (100 bp ladder).

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