Cargando…

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

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 fa...

Descripción completa

Detalles Bibliográficos
Autores principales: Wei, Jing, Li, Yanan, Cao, Yingli, Liu, Qi, Yang, Kankan, Song, Xiangjun, Shao, Ying, Qi, Kezong, Tu, Jian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9131491/
https://www.ncbi.nlm.nih.gov/pubmed/35646725
http://dx.doi.org/10.3389/fcimb.2022.879887
_version_ 1784713187371778048
author Wei, Jing
Li, Yanan
Cao, Yingli
Liu, Qi
Yang, Kankan
Song, Xiangjun
Shao, Ying
Qi, Kezong
Tu, Jian
author_facet Wei, Jing
Li, Yanan
Cao, Yingli
Liu, Qi
Yang, Kankan
Song, Xiangjun
Shao, Ying
Qi, Kezong
Tu, Jian
author_sort Wei, Jing
collection PubMed
description 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 × 10(2) 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.
format Online
Article
Text
id pubmed-9131491
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-91314912022-05-26 Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay Wei, Jing Li, Yanan Cao, Yingli Liu, Qi Yang, Kankan Song, Xiangjun Shao, Ying Qi, Kezong Tu, Jian Front Cell Infect Microbiol Cellular and Infection Microbiology 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 × 10(2) 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. Frontiers Media S.A. 2022-05-11 /pmc/articles/PMC9131491/ /pubmed/35646725 http://dx.doi.org/10.3389/fcimb.2022.879887 Text en Copyright © 2022 Wei, Li, Cao, Liu, Yang, Song, Shao, Qi and Tu https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Wei, Jing
Li, Yanan
Cao, Yingli
Liu, Qi
Yang, Kankan
Song, Xiangjun
Shao, Ying
Qi, Kezong
Tu, Jian
Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay
title Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay
title_full Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay
title_fullStr Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay
title_full_unstemmed Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay
title_short Rapid and Visual Detection of Porcine Parvovirus Using an ERA-CRISPR/Cas12a System Combined With Lateral Flow Dipstick Assay
title_sort rapid and visual detection of porcine parvovirus using an era-crispr/cas12a system combined with lateral flow dipstick assay
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9131491/
https://www.ncbi.nlm.nih.gov/pubmed/35646725
http://dx.doi.org/10.3389/fcimb.2022.879887
work_keys_str_mv AT weijing rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT liyanan rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT caoyingli rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT liuqi rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT yangkankan rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT songxiangjun rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT shaoying rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT qikezong rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay
AT tujian rapidandvisualdetectionofporcineparvovirususinganeracrisprcas12asystemcombinedwithlateralflowdipstickassay