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Jeon, Lee, and Song: Molecular Epidemiological Characteristics of Vibrio parahaemolyticus Isolated from Diarrheal Patients in Jeonnam, Korea
Original Article

J Bacteriol Virol 2009;39(3):229-235.

Published online: 18 January 2009

DOI: https://doi.org/10.4167/jbv.2009.39.3.229

Molecular Epidemiological Characteristics of Vibrio parahaemolyticus Isolated from Diarrheal Patients in Jeonnam, Korea

Doo Young Jeon1, Jeong Chi Lee2, Hyeon Je Song2

1Health and Environment Institute of Jeollanamdo, Gwangju, Korea

2Department of Clinical Pathology, Gwangju Health College University, Gwangju, Korea

∗Corresponding author: Hyeon-Je Song. Department of Clinical Pathology, Gwangju Health College University, 683-3 Sinchang-dong, Gwangsan-gu, Gwangju 506-701, Korea. Phone: +82-62-958-7622, e-mail: songha1@ghc.ac.kr

      Revised 6 July 200928 August 2009       Accepted 1 September 2009

Copyright © 2009 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

To investigate the occurrence and distribution of serotype, specific virulence genes, and pulse field gel electrophoresis (PFGE) patterns in Vibrio parahaemolyticus isolates from Jeonnam, Korea, we tested 87 strains which were identified with V. parahaemolyticus from diarrheal episode patients in 2005. In this study, 16 different O:K serotype combinations of V. parahaemolyticus were determined. The distributions of O and K serotypes were O4:K68 (51.72%), O1:K70 (18.39%), O3:K6 (5.74%), O1:K68 (4.60%) and O3:K57 (4.60%) respectively. Serotype O4:K68 was the regional dominant specific serotype of V. parahaemolyticus in Sinan of Jeonnam, Korea. For the detection of thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh) gene of V. parahaemolyticus, PCR was performed. The tdh gene was detected in all of the V. parahaemolyticus isolates from diarrheal patients, but trh gene was not detected. Analysis of PFGE patterns of 30 V. parahaemolyticus isolates showed 3 groups and 20 types. Among 14 O4:K68 serotypes which were isolated in Sinan, PFGE patterns of 12 strains were closely related (100%), but 2 strains were related by 58.3% and 45.4%, respectively. Also two strains of O1:K4 serotype in Gurye and two strains of O3:K6 serotype in Yeosu were closely related (100%), respectively. Although serotypes (O1:K4, O1:K70, O3:K6 and O4:K68) were different, PFGE patterns were related for more than 80.9%. Therefore, the epidemiological surveillance of V. parahaemolyticus is required by PFGE typing scheme as a further diagnostic tool.

Keywords: Vibrio parahaemolyticus, Serotyping, PFGE

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Figure 1.
PCR analysis of the tdh gene (198 bp) of the V. paraheamolyticus. M: molecular weight ladder (100 bp), lane 1: positive control (V. paraheamolyticus ATCC33816), lane 2: O1:K9, lane 3: O1:K13, lane 4: O1:K68, lane 5: O1:K70, lane 6:O2:K15, lane 7: O2:K19, lane 8: O3:K6, lane 9: O3:K57, lane 10:O4:K68, lane 11: O4:K68, lane 16: negative control.
jbv-39-229f1.tif
Figure 2.
PFGE patterns of enzyme (Not I) digested genomic DNA from V. parahaemolyticus isolates. M: Salmonella BAA-664, lane 1: O1:K4, lane 2: O1:K9, lane 3: O1:K13, lane 4: O1:K46, lane 5: O1:K56, lane 6: O1:K60, lane 7: O1:K61, lane 8: O1:K68, Lane 9: O1:K70, lane 10: O2:K15, lane 11: O2:K19, lane 12:O3:K6.
jbv-39-229f2.tif
Figure 3.
Dendrogram of 30 strains of V. parahaemolyticus isolates as generated by UPGMA method from genetic distances.
jbv-39-229f3.tif
Table 1.
Synthetic oligonucleotides used in this experiment
Target gene       5′-3′ DNA sequence of primer         Amplicon size (bps)
tdh F GGT ACT AAA TGG CTG ACA TCC 198
  R CCA AAT ACA TTT TAC TTG GAA  
trh F GGC TCA AAA TGG TTA AGC GCC 250
  R CAT TTC CGC TCT CAT ATG CTT  

F: forward primer, R: reverse primer

Table 2.
Occurrence and regional distribution of O and K serotype of V. parahaemolyticus isolated from diarrheal patients in Jeonnam, Korea
Serotype No. of strains
Total (%) Sinan Other regionsa
O1:K4 1 (1.15) 0 1
O1:K9 3 (3.45) 0 3
O1:K42 1 (1.15) 1 0
O1:K46 1 (1.15) 0 1
O1:K56 1 (1.15) 0 1
O1:K60 1 (1.15) 0 1
O1:K61 1 (1.15) 0 1
O1:K68 4 (4.60) 4 0
O1:K70 16 (18.39) 16 0
O2:K15 1 (1.15) 0 1
O2:K19 1 (1.15) 0 1
O3:K6 5 (5.74) 0 5
O3:K8 1 (1.15) 0 1
O3:K57 4 (4.60) 3 1
O4:K29 1 (1.15) 1 0
O4:K68 45 (51.72) 44 1
Total 87 69 18

a Yeosu, Hampyeong, Gurye, Boseong, Jangheung

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