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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...

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Autores principales: Wen, Li-bin, Wen, Shi-fu, He, Kong-wang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
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.
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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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