Frontiers in Cellular and Infection Microbiology (May 2022)

Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay

  • Jing Wei,
  • Jing Wei,
  • Yanan Li,
  • Yanan Li,
  • Yingli Cao,
  • Yingli Cao,
  • Qi Liu,
  • Qi Liu,
  • Kankan Yang,
  • Kankan Yang,
  • Xiangjun Song,
  • Xiangjun Song,
  • Ying Shao,
  • Ying Shao,
  • Kezong Qi,
  • Kezong Qi,
  • Jian Tu,
  • Jian Tu

DOI
https://doi.org/10.3389/fcimb.2022.879887
Journal volume & issue
Vol. 12

Abstract

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Porcine parvovirus (PPV) is one of the important causes of pig reproductive diseases. The most prevalent methods for PPV authentication are the polymerase chain reaction (PCR), enzyme-linked immunosorbent assay, and quantitative real-time PCR. However, these procedures have downsides, such as the fact that they take a long time and require expensive equipment. As a result, a rapid, visible, and economical clinical diagnostic strategy to detect PPV is necessary. In this study, three pairs of crRNA primers were designed to recognize the VP2 gene, and an ERA-CRISPR/Cas12a system for PPV detection was successfully developed. The approach involved isothermal detection at 37°C, and the method can be used for visual inspection. The detection limit of the ERA-CRISPR/Cas12a system was 3.75 × 102 copies/μL, and no cross reactions with other porcine viruses were found. In view of the preceding, a rapid, visible, and low-cost nucleic acid testing approach for PPV has been developed using the ERA-CRISPR/Cas12a system.

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