Genotyping of Vibrio parahaemolyticus by Infrequent Restriction Site Polymerase Chain Reaction.
- Author:
Dong G KEUM
1
;
Jung O KANG
;
Tae Y CHOI
Author Information
1. Department of Clinical Pathology, College of Medicine, Sungkygunwan University, Korea.
- Publication Type:Original Article
- Keywords:
Vibrio parahaemolyticus;
Infrequent restriction site-polymerase chain reaction;
Genotyping
- MeSH:
Diarrhea;
DNA;
DNA Fingerprinting;
Gwangju;
Humans;
Incheon;
Korea;
Polymerase Chain Reaction*;
Seoul;
Vibrio parahaemolyticus*;
Vibrio*
- From:Korean Journal of Clinical Microbiology
2002;5(2):119-123
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Infrequent restriction site PCR (IRS-PCR) is a recently described DNA fingerprinting technique based on selective amplification of restriction endonuclease-cleaved fragments. We applied of IRS-PCR to clinical isolates of Vibrio parahaemolyticus associated with diarrhea. METHODS: IRS-PCR assay was performed with adaptors for XbaI and HhaI restriction sites. A total of 35 strains of V. parahaemolyticus which were isolated from clinical specimens of patients with diarrhea were analyzed. The isolates were collected from different geographic areas of Seoul (n=12), Incheon (n=21) and Gwangju (n=2) during 1998-2000 in Korea. RESULTS: In IRS-PCR, amplifed DNA fragments between 50 and 400 bp were found to be the most reproducible in this study. When V. parahaemolyticus isolates were amplified with AH1 and PX-G as primers, 35 isolates could be grouped into five IRS-PCR patterns: A (n=16), B (n=4), C (n=6), D (n=5) and E (n=4). The patterns were subdivided into 15 subtypes: A1, A2, B1, B2, B3, B4, C1, C2, C3, D1, D2, D3, E1, E2 and E3. The IRS-PCR patterns of V. parahaemolyticus did not show any relationship with serotype or geographic origin, but the isolates from same outbreak produced a same pattern(A1). CONCLUSION: The results provide evidence of the discriminatory power of the IRS-PCR method as it applies to V. parahaemolyticus.
