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

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

doi:10.3389/fcimb.2022.879887

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

Front Cell Infect Microbiol. 2022 May 11:12:879887. doi: 10.3389/fcimb.2022.879887. eCollection 2022.

Authors

Jing Wei^{1

2}, Yanan Li^{1

2}, Yingli Cao^{1

2}, Qi Liu^{1

2}, Kankan Yang^{1

2}, Xiangjun Song^{1

2}, Ying Shao^{1

2}, Kezong Qi^{1

2}, Jian Tu^{1

2}

Affiliations

¹ Anhui Province Engineering Laboratory for Animal Food Quality and Bio-safety, College of Animal Science and Technology, Anhui Agricultural University, Hefei, China.
² Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, Anhui Agricultural University, Hefei, China.

Abstract

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.

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems
Parvovirus, Porcine* / genetics
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Swine
Swine Diseases* / genetics