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Immunocaptured-loop Mediated Isothermal Amplification Assay for Detection of Vibrio Parahaemolyticus in Seafood

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Abstract

Vibrio parahaemolyticus is an important seafood-borne pathogen associated with gastrointestinal disorders in humans. In this study, the efficacy of a loop-mediated isothermal amplification (LAMP) combined with immunocapture assay was firstly developed for detection of V. parahaemolyticus in pure culture, artificially and naturally contaminated seafood samples. The detection limit was 7.3 × 101 cfu/mL for pure culture and artificially contaminated samples by IC-LAMP, compared with 7.3 × 102 cfu/mL by LAMP. Furthermore, among 156 natural seafood samples, 13, 13 and 11 of samples were positive for V. arahaemolyticus by IC-LAMP, LAMP and culture-based method, respectively. V. parahaemolyticus was isolated from 11 of the 156 seafood samples by culture-based method, all of which were positive by IC-LAMP assay. A comparison between LAMP, IC-LAMP and culture-based method indicated that IC-LAMP was sensitive, specific and reliable for monitoring V. parahaemolyticus contamination in seafood samples.

Practical Application

Development of rapid, sensitive and specific methods for detection of V. parahaemolyticus is of utmost importance for monitoring and controlling contaminated seafood. In this study, loop-mediated isothermal amplification combined with the newly developed immunocapture showed higher detection limit than that by LAMP and more specific compared with that by culture-based method. The IC-LAMP was fit for monitoring V. parahaemolyticus contamination in seafood samples.

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