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Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus

African swine fever virus (ASFV), the etiological agent of African swine fever (ASF), a hemorrhagic fever of domestic pigs, has devastating consequences for the pig farming industry. More than 1,000,000 pigs have been slaughtered since 3 August 2018 in China. However, vaccines or drugs for ASF have...

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Autores principales: Miao, Faming, Zhang, Jingyuan, Li, Nan, Chen, Teng, Wang, Lidong, Zhang, Fei, Mi, Lijuan, Zhang, Jinxia, Wang, Shuchao, Wang, Ying, Zhou, Xintao, Zhang, Yanyan, Li, Min, Zhang, Shoufeng, Hu, Rongliang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6530510/
https://www.ncbi.nlm.nih.gov/pubmed/31156571
http://dx.doi.org/10.3389/fmicb.2019.01004
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author Miao, Faming
Zhang, Jingyuan
Li, Nan
Chen, Teng
Wang, Lidong
Zhang, Fei
Mi, Lijuan
Zhang, Jinxia
Wang, Shuchao
Wang, Ying
Zhou, Xintao
Zhang, Yanyan
Li, Min
Zhang, Shoufeng
Hu, Rongliang
author_facet Miao, Faming
Zhang, Jingyuan
Li, Nan
Chen, Teng
Wang, Lidong
Zhang, Fei
Mi, Lijuan
Zhang, Jinxia
Wang, Shuchao
Wang, Ying
Zhou, Xintao
Zhang, Yanyan
Li, Min
Zhang, Shoufeng
Hu, Rongliang
author_sort Miao, Faming
collection PubMed
description African swine fever virus (ASFV), the etiological agent of African swine fever (ASF), a hemorrhagic fever of domestic pigs, has devastating consequences for the pig farming industry. More than 1,000,000 pigs have been slaughtered since 3 August 2018 in China. However, vaccines or drugs for ASF have yet to be developed. As such, a rapid test that can accurately detect ASFV on-site is important to the timely implementation of control measures. In this study, we developed a rapid test that combines recombinase polymerase amplification (RPA) of the ASFV p72 gene with lateral flow detection (LFD). Results showed that the sensitivity of recombinase polymerase amplification with lateral flow dipstick (RPA-LFD) for ASFV was 150 copies per reaction within 10 min at 38°C. The assay was highly specific to ASFV and had no cross-reactions with other porcine viruses, including classical swine fever virus (CSFV). A total of 145 field samples were examined using our method, and the agreement of the positive rate between RPA-LFD (10/145) and real-time PCR (10/145) was 100%. Overall, RPA-LFD provides a novel alternative for the simple, sensitive, and specific identification of ASFV and showed potential for on-site ASFV detection.
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spelling pubmed-65305102019-05-31 Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus Miao, Faming Zhang, Jingyuan Li, Nan Chen, Teng Wang, Lidong Zhang, Fei Mi, Lijuan Zhang, Jinxia Wang, Shuchao Wang, Ying Zhou, Xintao Zhang, Yanyan Li, Min Zhang, Shoufeng Hu, Rongliang Front Microbiol Microbiology African swine fever virus (ASFV), the etiological agent of African swine fever (ASF), a hemorrhagic fever of domestic pigs, has devastating consequences for the pig farming industry. More than 1,000,000 pigs have been slaughtered since 3 August 2018 in China. However, vaccines or drugs for ASF have yet to be developed. As such, a rapid test that can accurately detect ASFV on-site is important to the timely implementation of control measures. In this study, we developed a rapid test that combines recombinase polymerase amplification (RPA) of the ASFV p72 gene with lateral flow detection (LFD). Results showed that the sensitivity of recombinase polymerase amplification with lateral flow dipstick (RPA-LFD) for ASFV was 150 copies per reaction within 10 min at 38°C. The assay was highly specific to ASFV and had no cross-reactions with other porcine viruses, including classical swine fever virus (CSFV). A total of 145 field samples were examined using our method, and the agreement of the positive rate between RPA-LFD (10/145) and real-time PCR (10/145) was 100%. Overall, RPA-LFD provides a novel alternative for the simple, sensitive, and specific identification of ASFV and showed potential for on-site ASFV detection. Frontiers Media S.A. 2019-05-15 /pmc/articles/PMC6530510/ /pubmed/31156571 http://dx.doi.org/10.3389/fmicb.2019.01004 Text en Copyright © 2019 Miao, Zhang, Li, Chen, Wang, Zhang, Mi, Zhang, Wang, Wang, Zhou, Zhang, Li, Zhang and Hu. http://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 Microbiology
Miao, Faming
Zhang, Jingyuan
Li, Nan
Chen, Teng
Wang, Lidong
Zhang, Fei
Mi, Lijuan
Zhang, Jinxia
Wang, Shuchao
Wang, Ying
Zhou, Xintao
Zhang, Yanyan
Li, Min
Zhang, Shoufeng
Hu, Rongliang
Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus
title Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus
title_full Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus
title_fullStr Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus
title_full_unstemmed Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus
title_short Rapid and Sensitive Recombinase Polymerase Amplification Combined With Lateral Flow Strip for Detecting African Swine Fever Virus
title_sort rapid and sensitive recombinase polymerase amplification combined with lateral flow strip for detecting african swine fever virus
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6530510/
https://www.ncbi.nlm.nih.gov/pubmed/31156571
http://dx.doi.org/10.3389/fmicb.2019.01004
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