Using loop-mediated isothermal amplification for visual detection of porcine parvovirus 7

Abstract

We developed and optimized a loop-mediated isothermal amplification (LAMP)-based method to detect porcine parvovirus 7 (PPV7). After using three pairs of specific primers to amplify PPV7 isothermally at 62 °C for 40 min, the amplified product was mixed with SYBR Green I, after which the sample turned green. The method detected PPV7 at concentrations as low as 40 copies/μL, and the sensitivity was consistent with that of nested polymerase chain reaction (PCR) analysis, which was tenfold higher than that of conventional PCR. No cross-reactivity occurred with porcine parvovirus 1, porcine circovirus type 3, porcine circovirus type 2, porcine pseudorabies virus, porcine epidemic diarrhea virus, or porcine reproductive and respiratory syndrome virus. Simultaneous analysis of 76 clinical samples was performed using LAMP, conventional PCR, and nested PCR. The results showed that our method is simple, rapid, sensitive, and specific for the rapid diagnosis of PPV7 in pig farms.

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Acknowledgements

We would like to thank Editage (www.editage.cn) for English language editing.

Funding

This work was supported financially by Anhui Provincial Primary Research & Developement Plan ( 202004f06020008, 201904f06020030) and Ningbo Health Branding Subject Fund (No. ppxk2018-10).

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Correspondence to Yongdong Li.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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All experiments were compliant with the ethical standards of Anhui Agricultural University.

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Wang, Y., Zhang, D., Bai, C. et al. Using loop-mediated isothermal amplification for visual detection of porcine parvovirus 7. 3 Biotech 11, 111 (2021). https://doi.org/10.1007/s13205-020-02623-5

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Keywords

  • Porcine parvovirus 7
  • Loop-mediated isothermal amplification
  • Visualization
  • Rapid detection