Cargando…
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...
Autores principales: | , , , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1783420673228537856 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6530510 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT miaofaming rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT zhangjingyuan rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT linan rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT chenteng rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT wanglidong rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT zhangfei rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT milijuan rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT zhangjinxia rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT wangshuchao rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT wangying rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT zhouxintao rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT zhangyanyan rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT limin rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT zhangshoufeng rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus AT hurongliang rapidandsensitiverecombinasepolymeraseamplificationcombinedwithlateralflowstripfordetectingafricanswinefevervirus |