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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-11-01
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| Series: | BMC Veterinary Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12917-021-03060-z |
| _version_ | 1818856608052019200 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-19T08:27:12Z |
| format | Article |
| id | doaj.art-dadee292ef4544c98b82f0180840890f |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-19T08:27:12Z |
| publishDate | 2021-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-dadee292ef4544c98b82f0180840890f2022-12-21T20:29:17ZengBMCBMC Veterinary Research1746-61482021-11-0117111210.1186/s12917-021-03060-zEstablishment of a peptide-based enzyme-linked immunosorbent assay for detecting antibodies against PRRSV M proteinJun Zhao0Rubo Zhang1Ling Zhu2Huidan Deng3Fengqing Li4Lei Xu5Jianbo Huan6Xiangang Sun7Zhiwen Xu8College of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityCollege of Veterinary Medicine, Sichuan Agricultural UniversityAbstract 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.https://doi.org/10.1186/s12917-021-03060-zPorcine reproductive and respiratory syndrome virusM proteinSynthetic peptideEnzyme-linked immunosorbent assayNADC30-like PRRSV inactivated vaccinePig |
| spellingShingle | 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 BMC Veterinary Research Porcine reproductive and respiratory syndrome virus M protein Synthetic peptide Enzyme-linked immunosorbent assay NADC30-like PRRSV inactivated vaccine Pig |
| title | 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_short | 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 |
| topic | Porcine reproductive and respiratory syndrome virus M protein Synthetic peptide Enzyme-linked immunosorbent assay NADC30-like PRRSV inactivated vaccine Pig |
| url | https://doi.org/10.1186/s12917-021-03060-z |
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