Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein

Abstract Background Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, t...

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Autores principales:	Jun Zhao, Rubo Zhang, Ling Zhu, Huidan Deng, Fengqing Li, Lei Xu, Jianbo Huan, Xiangang Sun, Zhiwen Xu
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Porcine reproductive and respiratory syndrome virus M protein Synthetic peptide Enzyme-linked immunosorbent assay NADC30-like PRRSV inactivated vaccine Pig Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/dadee292ef4544c98b82f0180840890f
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spelling	oai:doaj.org-article:dadee292ef4544c98b82f0180840890f2021-11-21T12:25:53ZEstablishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein10.1186/s12917-021-03060-z1746-6148https://doaj.org/article/dadee292ef4544c98b82f0180840890f2021-11-01T00:00:00Zhttps://doi.org/10.1186/s12917-021-03060-zhttps://doaj.org/toc/1746-6148Abstract Background Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, the B cell line epitopes of PRRSV M protein were predicted, and two peptide ELISA assays were established (M-A110-129 ELISA, M-A148-174 ELISA) to detect antibodies against PRRSV M protein. Field serum samples collected from pig farms were used to validate the peptide ELISA and compare it with an indirect immunofluorescence assay. Results The sensitivity and specificity of M-A110-129 ELISA and M-A148-174 ELISA were (111/125) 88.80%, (69/70) 98.57% and (122/125) 97.60%, (70/70) 100%, relative to indirect immunofluorescence assay. This peptide ELISA could detect antibodies against different genotypes of PRRSV including type 1 PRRSV, classical PRRSV, HP-PRRSV, and NADC30 like PRRSV, but not antibodies against other common swine viruses. The results of ROC analysis showed that the area under the curve (AUC) of the M-A110-129 ELISA and M-A148-174 ELISA were 0.967 and 0.996, respectively. Compared the concordance of results using two peptide ELISA assays, the IDEXX PRRSV X3 Ab ELISA and a virus neutralization test, were assessed using a series of 147 sera from pigs vaccinated with the NADC30-like PRRSV inactivated vaccine. The M-A148-174 ELISA had the best consistency, with a Cohen’s kappa coefficient of 0.8772. The concordance rates of the Hipra PRRSV ELISA kit, M-A110-129 ELISA and M-A148-174 ELISA in the field seropositive detection results were 91.08, 86.32 and 95.35%, relative to indirect immunofluorescence assay. Conclusions In summary, compared with M-A110-129 ELISA, the PRRSV M-A148-174 ELISA is of value for detecting antibodies against PRRSV and the evaluation of the NADC30-like PRRSV inactivated vaccine, but the advantage is insufficient in serological early diagnosis.Jun ZhaoRubo ZhangLing ZhuHuidan DengFengqing LiLei XuJianbo HuanXiangang SunZhiwen XuBMCarticlePorcine reproductive and respiratory syndrome virusM proteinSynthetic peptideEnzyme-linked immunosorbent assayNADC30-like PRRSV inactivated vaccinePigVeterinary medicineSF600-1100ENBMC Veterinary Research, Vol 17, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Porcine reproductive and respiratory syndrome virus M protein Synthetic peptide Enzyme-linked immunosorbent assay NADC30-like PRRSV inactivated vaccine Pig Veterinary medicine SF600-1100
spellingShingle	Porcine reproductive and respiratory syndrome virus M protein Synthetic peptide Enzyme-linked immunosorbent assay NADC30-like PRRSV inactivated vaccine Pig Veterinary medicine SF600-1100 Jun Zhao Rubo Zhang Ling Zhu Huidan Deng Fengqing Li Lei Xu Jianbo Huan Xiangang Sun Zhiwen Xu Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
description	Abstract Background Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry globally. Evaluation of antibody responses and neutralizing antibody titers is the most effective method for vaccine evaluation. In this study, the B cell line epitopes of PRRSV M protein were predicted, and two peptide ELISA assays were established (M-A110-129 ELISA, M-A148-174 ELISA) to detect antibodies against PRRSV M protein. Field serum samples collected from pig farms were used to validate the peptide ELISA and compare it with an indirect immunofluorescence assay. Results The sensitivity and specificity of M-A110-129 ELISA and M-A148-174 ELISA were (111/125) 88.80%, (69/70) 98.57% and (122/125) 97.60%, (70/70) 100%, relative to indirect immunofluorescence assay. This peptide ELISA could detect antibodies against different genotypes of PRRSV including type 1 PRRSV, classical PRRSV, HP-PRRSV, and NADC30 like PRRSV, but not antibodies against other common swine viruses. The results of ROC analysis showed that the area under the curve (AUC) of the M-A110-129 ELISA and M-A148-174 ELISA were 0.967 and 0.996, respectively. Compared the concordance of results using two peptide ELISA assays, the IDEXX PRRSV X3 Ab ELISA and a virus neutralization test, were assessed using a series of 147 sera from pigs vaccinated with the NADC30-like PRRSV inactivated vaccine. The M-A148-174 ELISA had the best consistency, with a Cohen’s kappa coefficient of 0.8772. The concordance rates of the Hipra PRRSV ELISA kit, M-A110-129 ELISA and M-A148-174 ELISA in the field seropositive detection results were 91.08, 86.32 and 95.35%, relative to indirect immunofluorescence assay. Conclusions In summary, compared with M-A110-129 ELISA, the PRRSV M-A148-174 ELISA is of value for detecting antibodies against PRRSV and the evaluation of the NADC30-like PRRSV inactivated vaccine, but the advantage is insufficient in serological early diagnosis.
format	article
author	Jun Zhao Rubo Zhang Ling Zhu Huidan Deng Fengqing Li Lei Xu Jianbo Huan Xiangang Sun Zhiwen Xu
author_facet	Jun Zhao Rubo Zhang Ling Zhu Huidan Deng Fengqing Li Lei Xu Jianbo Huan Xiangang Sun Zhiwen Xu
author_sort	Jun Zhao
title	Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
title_short	Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
title_full	Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
title_fullStr	Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
title_full_unstemmed	Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein
title_sort	establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against prrsv m protein
publisher	BMC
publishDate	2021
url	https://doaj.org/article/dadee292ef4544c98b82f0180840890f
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Establishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M protein

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