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

Yong-Kyun Kim; Jae-Seok Kim; Han-Sung Kim; Wonkeun Song; Hyoun Chan Cho; Kyu Man Lee

doi:10.3343/kjlm.2008.28.4.286

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Kim, Kim, Kim, Song, Cho, and Lee: Molecular Typing of Staphylococcus aureus Isolated from Blood on the Basis of Coagulase Gene Polymorphism and Toxin Genes

Original Article

Korean J Leg Med 2008;28(4):286-292.

Published online: 14 February 2008

DOI: https://doi.org/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

Yong-Kyun Kim, M.D., Jae-Seok Kim, M.D., Han-Sung Kim, M.D., Wonkeun Song, M.D., Hyoun Chan Cho, M.D., Kyu Man Lee, M.D.

Department of Laboratory Medicine, Hallym University College of Medicine, Seoul, Korea

Received 14 January 20089 May 2008 Accepted 27 May 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Staphylococcus aureus, Coagulase, Restriction fragment length polymorphism, Enterotoxins, Methicillin resistance

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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).

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).

Table 1.

A number of 120 S. aureus blood isolates according to RFLP patterns, staphylococcal toxic genes, mecA gene, and methicillin resistance

PCR-amplified coa gene size^∗	RFLP pattern^†	N	Toxin gene	mecA gene	MRSA (N=85)	MSSA (N=35)
S (N=6)	S1	1	sec, tst	+	1
	S2	1	-	+	1
			tst	-		1
	S3	4	sea, tst	-		2
			sea, see, tst	-		1
M (N=13)	M1	1	-	-		1
	M2	1	-	-		1
	M3	1	-	-		1
	M4	2	-	-		2
			-	-		1
	M5	8	sea	-		3
			see	-		1
			sea, see	-		3
L (N=97)	L1	37	-	-		1
			-	+	4
			sea	+	16
			sea, see	+	16
	L2	3	-	+	2
			sec, tst	+	1
	L3	3	sea	-		1
			sea, see	-		2
	L4	1	sec, tst	+	1
	L5	46	-	-		1
			sea	-		1
			sea, see	-		1
			-	+	2
			tst	+	6
			sea, sec	+	1
			sec, tst	+	32
			sea, sec, tst	+	2
	L6	4	-	-		4
	L7	3	-	-		1
			sec	-		2
XL (N=4)	XL1	4	-	-		4

^∗ , The categorization of size of coa gene is based on the size reported by Ishino et al.[10];

^† , The coa gene RFLP patterns are arbitrarily categorized in this study.

Abbreviations: RFLP, restriction fragment length polymorphism; MRSA, methicillin-resistant Staphylococcus aureus; MSSA, methicillin-susceptible S. aureus.

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