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Kim, Hwang, and Chang: Correlation between Sau1 Restriction and Modification Complex Type and Coagulase Serotype or SCCmec Type of Staphylococcus aureus
Original Article

J Bacteriol Virol 2010;40(4):163-170.

Published online: 18 January 2010

DOI: https://doi.org/10.4167/jbv.2010.40.4.163

Correlation between Sau1 Restriction and Modification Complex Type and Coagulase Serotype or SCCmec Type of Staphylococcus aureus

So Yeon Kim, Soo Myung Hwang, Kyung Soo Chang∗

Department of Clinical Laboratory Science, Catholic University of Pusan, Busan, Korea

∗Corresponding author: Kyung Soo Chang. Department of Clinical Laboratory Science, Catholic University of Pusan, Busan 609-757, Korea. Phone: +82-51-510-0565, Fax: +82-51-510-0568 e-mail: kschang@cup.ac.kr

∗∗ This research was supported by the research fund of the Catholic University of Pusan.

      Revised 3 September 201023 September 2010       Accepted 29 September 2010

Copyright © 2010 Korean Society for Legal Medicine

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

Staphylococcus aureus coagulase serotype I to VIII isolated from clinical samples could be classified into two groups, methicillin-sensitive S. aurues (MSSA) and methicilln-resistant S. aurues (MRSA), by antibiotics susceptibility and existence of mecA which is a gene related with methicillin resistance. Coagulase serotype I, VI, and VIII were MSSA which showed different antimicrobial susceptibility. Coagluase serotype II-V and VII were MRSA in which mecA and SCCmec were detected. To analyze Sau1 restriction and modification (R-M) complex types by coagulase type and SCCmec type, sau1hsdR, sau1hsdM and sau1hsdS genes involved in Sau1 R-M complex were detected by PCR, we found five complex types such as M1, R2M2, R2M2, R2M2S1, and R2M2S2. Coagulase serotype I, VI, and VIII of MSSA were M1, R2M2 and R2M2, respectively. SCCmec type II and coagulase serotype II, SCCmec type III and coagulase serotype III, SCCmec type IV and coagulase serotype V, and SCCmec type IV and coagulase serotype IV, VII of MRSA were Sau1 R-M complex type R2M2S1, R2M2, R2M2, and R2M2S2, respectively. Taken together, correlation between Sau1 R-M complex types and coagulase or SCCmec types of S. aureus was found.

Keywords: Staphylococcus aureus, Sau1 restriction-modification (R-M) complex type, Coagulase serotype, SCCmec type, hsd gene

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Figure 1.
Agarose gel electrophoresis of DNA fragment amplified from the SCCmec (locus A, B, C, D, E, F, G, H) by PCR with specific primers. Lane M: the marker (100 bp plus DNA ladder), lane C: S. aureus (ATCC25923), the molecular size of locus A to H are 495 bp, 284 bp, 209 bp, 342 bp, 243 bp, 414 bp, 381 bp and 303 bp, respectively.
jbv-40-163f1.tif
Figure 2.
Agarose gel electrophoresis of DNA fragment amplified from the Sau1 (hsdM, hsdR, hsdS) gene by PCR with specific primers. Lane M, 100 bp plus DNA ladder marker, lane C: S. aureus (ATCC25943), the molecular size of hsdM1, M2, S1, S2, R1 and R2 are 1,577 bp, 1,535 bp, 2,810 bp, 258 bp, 327 bp and 1,220 bp, respectively.
jbv-40-163f2.tif
Figure 3.
Genetic maps of type 1 Restriction and Modification system of S. aureus (A). S. aureus (USA_TCH1516) strain, (B). S. aureus (COL) strain.
jbv-40-163f3.tif
Table 1.
Primers and predicted sizes of PCR products for SCCmec type and Sau1 R-M complex genes
Primer names   Primer sequence (5′ to 3′) Amplified sizes Target gene
Primers for SCCmec type       SCCmec type
A CIF2 F2 TTCGAGTTGCTGATGAAGAAGG 495 I
  CIF2 R2 ATTTACCACAAGGACTACCAGC    
B KDP F1 AATCATCTGCCATTGGTGATGC 284 II
  KDP R1 CGAATGAAGTGAAAGAAAGTGG    
C MECI P2 ATCAAGACTTGCATTCAGGC 209 II, III
  MECI P3 GCGGTTTCAATTCACTTGTC    
D DCS F2 CATCCTATGATAGCTTGGTC 342 I, II, IV
  DCS R1 CTAAATCATAGCCATGACCG    
E RIF4 F3 GTGATTGTTCGAGATATGTGG 243 III
  RIF4 R9 CGCTTTATCTGTATCTATCGC    
F RIF5 F10 TTCTTAAGTACACGCTGAATCG 414 III
  RIF5 R13 GTCACAGTAATTCCATCAATGC    
G IS431 P4 CAGGTCTCTTCAGATCTACG 381 IA
  pUB110 R1 GAGCCATAAACACCAATAGCC    
H IS431 P4 CAGGTCTCTTCAGATCTACG 303 IIIA
  pT181 R1 GAAGAATGGGGAAAGCTTCAC    
Primer for Sau1 R-M complex genes       Sau1 gene type
hsdM1-FL   TTA GCT TCA AAG ATG GCA GT 1,577 Sau1hsdM1
hsdM1-RL   GGG AAC CTC AAC TCT GGC AC    
hsdM2-F   CCT GGG AAT CTC AAC TCT GG 1,535 Sau1hsdM2
hsdM2-R   GAC ATT CAC CCA ATC CTG AC    
hsdR1-F   CGA AAC AAG CGT ATT GTT GC 258 Sau1hsdR1
hsdR1-R   TGC TCT ACA ACT TGT TCC AC    
hsdR2-F   GCG TTG ATT GTA TTC GGG AC 2,810 Sau1hsdR2
hsdR2-R   CCT ACA CTA ATC TGA CGA GG    
hsdS1-F   CAA CGG CAT ACC ATT TTT AAG 327 Sau1hsdS1
hsdS1-R   AGA CCT TCT CGA CTA CCT C    
hsdS2-F   CAA TTA ATA GGT TGT TAT CA 1,220 Sau1hsdS2
hsdS2-R   ATG CAT ACC TGA AAG AAC TT    

Reference, Cha, et al. (7), Waldron, et al. (18)

Table 2.
CLSI disk diffusion antimicrobial susceptibility of S. aureus isolates according to coagulase types
Antimicrobial disk Coagulase type
I II III IV V VI VII VIII
Penicillin S or R R R R R R R R
Methicillin S R R R R S R S
Cefoxitin S R R R S or R S R S
Vancomycin S S S S S S S S
Teicoplanin S S S S S S S S

R: resistant, S: susceptible

Table 3.
Summary of relationship of Sau1 R-M type, coagulase type, SCCmec type, and methicillin susceptibility
Types Sau1 R-M type
M1 R2M2 R2M2 R2M2S1 R2M2S2
Coagulase type I VI, VIII III, V II IV, VII
SCCmec type ND ND III, IV II IV
Methicillin type MSSA MSSA MRSA MRSA MRSA
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