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Rapid visual detection of Vibrio parahaemolyticus in seafood samples by loop-mediated isothermal amplification with hydroxynaphthol blue dye

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Abstract

The detection and monitoring of Vibrio parahaemolyticus pathogen in aquatic foods have become essential for preventing outbreaks. In this study, loop-mediated isothermal amplification (LAMP) assay with the azo dye, hydroxynaphthol blue (HNB) was developed targeting species-specific tlh gene. The assay was carried out on 62 seafood samples that included clam and shrimp and compared with conventional LAMP assay performed with the commonly used fluorescent dye, conventional PCR, and real-time PCR (RT-PCR). Of 62 samples studied for tlh gene, 32 (51.61%) gave positive by HNB-LAMP, which comprised 22 (70.96%) clam samples and 10 (32.25%) shrimp samples. The HNB-LAMP assay was found to be highly sensitive, specific, and superior to conventional PCR (p > 0.05). RT-PCR presented higher sensitivity than HNB-LAMP; however, it has the limitation of being cost-intensive and requiring technical expertise to perform. HNB-LAMP is affordable, rapid, simple, and easy to perform, allowing naked eye visualization.

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Acknowledgements

The financial support by the Department of Biotechnology, Government of India through the Project (BT/PR12230/AAQ/3/698/2014) is greatly acknowledged.

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Authors and Affiliations

  1. Division of Infectious Diseases, Nitte University Centre for Science Education and Research, Paneer Campus, Deralakatte, Mangalore, 575018, India

    Karanth Padyana Anupama, Ashwath Nayak, Indrani Karunasagar & Biswajit Maiti

  2. Nitte (Deemed To Be University), University Enclave, Medical Sciences Complex, Deralakatte, Mangalore, 575018, India

    Indrani Karunasagar

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  1. Karanth Padyana Anupama
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  2. Ashwath Nayak
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Anupama, K.P., Nayak, A., Karunasagar, I. et al. Rapid visual detection of Vibrio parahaemolyticus in seafood samples by loop-mediated isothermal amplification with hydroxynaphthol blue dye. World J Microbiol Biotechnol 36, 76 (2020). https://doi.org/10.1007/s11274-020-02851-0

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