期刊
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
卷 145, 期 1, 页码 244-249出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.ijfoodmicro.2010.12.030
关键词
Vibrio parahaemolyticus; Thermostable direct hemolysin (TDH); TDH-related hemolysin (TRH); Monoclonal antibody (mAb); Enzyme-linked immunosorbent assay (ELISA)
资金
- Department of Biotechnology Govt. of India
Thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) are considered important virulence factors of Vibrio parahaemolyticus and strains producing either of these or both are considered pathogenic. In this study, we generated monoclonal antibodies (mAbs) against purified TRH recombinant protein of pathogenic V. parahaemolyticus. Sandwich enzyme-linked immunosorbent assays (ELISA) using the hybridoma clone 4810 showed higher sensitivity of detection compared to other clones. Using mAb 4B10 based sandwich ELISA, we could detect pathogenic V. parahaemolyticus in 41.18% (14 out of 34) of the seafood samples analyzed. PCR targeting the toxR gene showed the presence of V. parahaemolyticus in 64.7% (22 out of 34) seafood samples. Further. PCR targeting the virulence genes showed that 6 seafood samples harboured the tdh gene while 9 harboured the trh gene indicating the presence of pathogenic V. parahaemolyticus. Our results show that mAb 4810 sandwich ELISA developed in this study could be used as a rapid method for screening seafood samples for the presence of pathogenic V. parahaemolyticus. (C) 2011 Elsevier B.V. All rights reserved.
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