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Development of a rapid loop-mediated isothermal amplification (LAMP) assay for visual detection of porcine parvovirus (PPV) and its application

  • Bacterial, Fungal and Virus Molecular Biology - Research Paper
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Abstract

Porcine parvovirus (PPV) infection is one of the most important causes of reproductive failure in pigs impacting the piggery industry globally with huge economic losses. A cost-effective, simple, rapid, specific, and sensitive method is critical for monitoring PPV infection on pig farms. The main aim of the present study was to develop and evaluate a loop-mediated isothermal amplification (LAMP) assay for rapid visual detection of porcine parvovirus (PPV) in pigs. A set of six LAMP primers including two outer primers, two inner primers, and two loop primers were designed utilizing the conserved region of capsid protein VP2 gene sequences of PPV and was applied for detection of PPV from porcine samples. Time and temperature conditions for amplification of PPV genes were optimized to be 30 min at 63 °C. The developed assay was ten-fold more sensitive than conventional PCR with analytical sensitivity of 20 pg and 200 pg, respectively. This is the first report of detection of PPV by LAMP assay from India. The assay did not cross-react with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), or classical swine fever virus (CSFV). The LAMP assay was assembled into a LAMP assay kit of 20 reactions and was validated in different laboratories in India. The newly developed LAMP assay was proved to be a specific, sensitive, rapid, and simple method for visual detection of PPV which does not require even costly equipments for performing the test. It complements and extends previous methods for PPV detection and provides an alternative approach for detection of PPV.

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Funding

This work was supported by funding from DBT, Govt. of India in the form of ADMaC project.

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Authors and Affiliations

  1. Animal Health Laboratory, Indian Council of Agricultural Research – National Research Centre On Pig (ICAR-NRC On Pig), Rani, Guwahati, 781131, Assam, India

    S. Rajkhowa, M. Choudhury & S. R. Pegu

  2. Department of Microbiology, AAU, Guwahati, India

    D. K. Sarma

  3. Division of Veterinary Microbiology & Immunology, SKUAST-Kashmir, Srinagar, India

    I. Hussain

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  1. S. Rajkhowa
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  2. M. Choudhury
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  3. S. R. Pegu
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  4. D. K. Sarma
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  5. I. Hussain
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Correspondence to S. Rajkhowa.

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Rajkhowa, S., Choudhury, M., Pegu, S.R. et al. Development of a rapid loop-mediated isothermal amplification (LAMP) assay for visual detection of porcine parvovirus (PPV) and its application . Braz J Microbiol 52, 1725–1732 (2021). https://doi.org/10.1007/s42770-021-00569-1

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  • DOI: https://doi.org/10.1007/s42770-021-00569-1

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