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Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1
BACKGROUND: Porcine circovirus-like virus P1 is a newly discovered virus. To date, there has been no specific serological assay for use in the diagnosis of P1 infection. RESULTS: Because P1 has high homology to porcine circovirus type 2 (PCV2) at the nucleotide level, the C-terminal portion of the c...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4717539/ https://www.ncbi.nlm.nih.gov/pubmed/26786046 http://dx.doi.org/10.1186/s12917-016-0641-8 |
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author | Wen, Li-bin Wen, Shi-fu He, Kong-wang |
author_facet | Wen, Li-bin Wen, Shi-fu He, Kong-wang |
author_sort | Wen, Li-bin |
collection | PubMed |
description | BACKGROUND: Porcine circovirus-like virus P1 is a newly discovered virus. To date, there has been no specific serological assay for use in the diagnosis of P1 infection. RESULTS: Because P1 has high homology to porcine circovirus type 2 (PCV2) at the nucleotide level, the C-terminal portion of the capsid protein (amino acids 73–114), a discriminative antigen, was expressed in a prokaryotic expression system. The recombinant product (rctCap), composed of three identical repeated domains, was shown to be strongly immunoreactive to P1-specific serum. This assay was validated by comparison with an indirect immunofluorescence assay (IFA). The diagnostic sensitivity and specificity of the rctCap enzyme-linked immunosorbent assay (ELISA) developed in this study are 93.6 % and 98.3 %, respectively, compared with the results from IFAs on 450 sera samples from pigs. CONCLUSIONS: The indirect ELISA that we developed with rctCap, the recombinant capsid fragment containing the 217–342 nt repeat domain, was sensitive, specific, and suitable for the large-scale detection of P1 infections in swine. |
format | Online Article Text |
id | pubmed-4717539 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-47175392016-01-20 Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 Wen, Li-bin Wen, Shi-fu He, Kong-wang BMC Vet Res Methodology Article BACKGROUND: Porcine circovirus-like virus P1 is a newly discovered virus. To date, there has been no specific serological assay for use in the diagnosis of P1 infection. RESULTS: Because P1 has high homology to porcine circovirus type 2 (PCV2) at the nucleotide level, the C-terminal portion of the capsid protein (amino acids 73–114), a discriminative antigen, was expressed in a prokaryotic expression system. The recombinant product (rctCap), composed of three identical repeated domains, was shown to be strongly immunoreactive to P1-specific serum. This assay was validated by comparison with an indirect immunofluorescence assay (IFA). The diagnostic sensitivity and specificity of the rctCap enzyme-linked immunosorbent assay (ELISA) developed in this study are 93.6 % and 98.3 %, respectively, compared with the results from IFAs on 450 sera samples from pigs. CONCLUSIONS: The indirect ELISA that we developed with rctCap, the recombinant capsid fragment containing the 217–342 nt repeat domain, was sensitive, specific, and suitable for the large-scale detection of P1 infections in swine. BioMed Central 2016-01-19 /pmc/articles/PMC4717539/ /pubmed/26786046 http://dx.doi.org/10.1186/s12917-016-0641-8 Text en © Wen et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Wen, Li-bin Wen, Shi-fu He, Kong-wang Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 |
title | Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 |
title_full | Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 |
title_fullStr | Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 |
title_full_unstemmed | Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 |
title_short | Development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated Cap protein for the diagnosis of porcine circovirus-like virus P1 |
title_sort | development and application of an indirect enzyme-linked immunosorbent assay using recombinant truncated cap protein for the diagnosis of porcine circovirus-like virus p1 |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4717539/ https://www.ncbi.nlm.nih.gov/pubmed/26786046 http://dx.doi.org/10.1186/s12917-016-0641-8 |
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