4.7 Article

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

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FRONTIERS MEDIA SA
DOI: 10.3389/fcimb.2022.879887

关键词

CRISPR-Cas12a; enzymatic recombinase amplification; porcine parvovirus; lateral flow dipstick; rapid detection

资金

  1. Application of supporting technology and poverty alleviation demonstration of ecological breeding of native black pig in contiguous poverty-stricken area of Dabie Mountains, Anhui [201907d06020016]
  2. University Excellent Talents Support Program [gxyqZD2020009]

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A rapid, visible, and low-cost nucleic acid testing approach for porcine parvovirus (PPV) has been developed using the ERA-CRISPR/Cas12a system.
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 degrees C, and the method can be used for visual inspection. The detection limit of the ERA-CRISPR/Cas12a system was 3.75 x 10(2) copies/mu L, 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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