Korean J Clin Pathol.  2001 Oct;21(5):355-359.

Detection of Thermostable Direct Hemolysin Gene and Thermostable Direct Hemolysin-Related Hemolysin Gene of Vibrio parahaemolyticus by Polymerase Chain Reaction

• Kim WJ
• Keum DG
• Soe YH
• Choi TY

Abstract

BACKGROUND
Vibrio parahaemolyticus is one of the most important food-borne pathogens in Korea. Although the mechanism of its pathologic effect is still not clearly understood, epidemiological studies have suggested a very strong association of thermostable direct hemolysin (TDH) and thermostable direct hemolysin-related hemolysin (TRH) with the disease. We detected the tdh gene and the trh gene of V. parahaemolyticus isolates by polymerase chain reaction (PCR).
METHODS
V. parahaemolyticus strains were isolated from clinical specimens of patients with diarrhea in different geographic areas of Seoul (16 cases), Inchon (27 cases) and Kwang Joo (2 cases) in Korea between 1998 and 2000. The colonies selected were identified by using the API 20E identification strip (API system, Analytab Products, Montalieu-Vercieu, France). PCR protocols were established for specific detection of the tdh and trh genes. Oligonucleotide primers were designed based on the reported nucleotide sequence of the tdh2 gene and the trh1 gene, respectively.
RESULTS
The protocols established for the tdh and trh genes could detect 400 fg (100 colony-forming units) of cellular DNA carrying the respective gene. All strains of V. parahaemolyticus isolates in this study contained the tdh gene but not the trh gene.
CONCLUSIONS
The detection of the tdh gene and trh gene of V. parahaemolyticus using the PCR is an easy and rapid method.

Keyword

Vibrio parahaemolyticus; tdh gene; trh gene; PCR

MeSH Terms

Base Sequence
Diarrhea
DNA
DNA Primers
Humans
Incheon
Korea
Polymerase Chain Reaction*
Seoul
Vibrio parahaemolyticus*
Vibrio*
DNA
DNA Primers