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. 2012 Apr 4;3(3):204–212. doi: 10.1007/s13238-012-2017-6

Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood

Bin Liu 1, Xiaohua He 1, Wanyi Chen 1, Shuijing Yu 1, Chunlei Shi 1, Xiujuan Zhou 1, Jing Chen 1, Dapeng Wang 1, Xianming Shi 1,✉
PMCID: PMC4875422  PMID: 22477699

Abstract

A real time PCR assay for the detection of Vibrio parahaemolyticus in seafood samples was developed using a novel specific target and a competitive internal amplification control (IAC). The specificity of this assay was evaluated using 390 bacterial strains including V. parahaemolyticus, and other strains belonging to Vibrio and non-Vibrio species. The real time PCR assay unambiguously distinguished V. parahaemolyticus with a detection sensitivity of 4.8 fg per PCR with purified genomic DNA or 1 CFU per reaction by counting V. parahaemolyticus colonies. The assays of avoiding interference demonstrated that, even in the presence of 2.1 μg genomic DNA or 107 CFU background bacteria, V. parahaemolyticus could still be accurately detected. In addition, the IAC was used to indicate false-negative results, and lower than 94 copies of IAC per reaction had no influence on the detection limit. Ninety-six seafood samples were tested, of which 58 (60.4%) were positive, including 3 false negative results. Consequently, the real time PCR assay is effective for the rapid detection of V. parahaemotyticus contaminants in seafood.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s13238-012-2017-6 and is accessible for authorized users.

Keywords: Vibrio parahaemolyticus, real time PCR, internal amplification control, seafood

Electronic supplementary material

13238_2012_2017_MOESM1_ESM.pdf (87.8KB, pdf)

Supplementary material, approximately 87.7 KB.

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13238_2012_2017_MOESM1_ESM.pdf (87.8KB, pdf)

Supplementary material, approximately 87.7 KB.

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