AUTHOR=Wang Yi , Li Hui , Li Dongxun , Li Kewei , Wang Yan , Xu Jianguo , Ye Changyun 

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

JOURNAL=Frontiers in Microbiology

VOLUME=7

YEAR=2016

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2016.02047

DOI=10.3389/fmicb.2016.02047

ISSN=1664-302X

ABSTRACT=<p><italic>Vibrio parahaemolyticus</italic> (<italic>V. parahaemolyticus</italic>) 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 <italic>toxR</italic> gene of <italic>V. parahaemolyticus</italic> 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 <italic>V. parahaemolyticus</italic> DNA and from approximately 4.2 × 10<sup>2</sup> CFU in 1 mL of oyster homogenate. No cross-reaction with other <italic>Vibrio</italic> species and with non-<italic>Vibrio</italic> species was observed. Therefore, the MCDA-LFB method established in the current report is suitable for the rapid screening of <italic>V. parahaemolyticus</italic> in clinical, food, and environmental samples.</p>