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Vibrio parahaemolyticus  ( V. parahaemolyticus ) is a marine seafood-borne pathogen causing severe illnesses in humans and aquatic animals. In the present study, multiple cross displacement amplification was combined with a lateral flow biosensor (MCDA-LFB) to detect the  toxR  gene of  V. parahaemolyticus  in DNA extracts from pure cultures and spiked oyster homogenates. Amplification was carried out at a constant temperature (62°C) for only 30 min, and amplification products were directly applied to the biosensor. The entire process, including oyster homogenate processing (30 min), isothermal amplification (30 min) and results indicating (∼2 min), could be completed within 65 min. Amplification product was detectable from as little as 10 fg of pure  V. parahaemolyticus  DNA and from approximately 4.2 × 10 2  CFU in 1 mL of oyster homogenate. No cross-reaction with other  Vibrio  species and with non- Vibrio  species was observed. Therefore, the MCDA-LFB method established in the current report is suitable for the rapid screening of  V. parahaemolyticus  in clinical, food, and environmental samples.

Multiple Cross Displacement Amplification Combined with Gold Nanoparticle-Based Lateral Flow Biosensor for Detection of Vibrio parahaemolyticus