Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus
Abstract Porcine reproductive and respiratory syndrome virus (PRRSV) infection causes significant economic loss to the global pig industry. Genotype 1 and 2 PRRSV (PRRSV-1 and -2) infections have been reported in China, Europe and America. For accurate prevention, nanobodies were first used as diagn...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-03-01
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| Series: | Animal Diseases |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s44149-024-00114-1 |
| _version_ | 1797233667672637440 |
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| author | Xu Chen Yueting Chang Lu Zhang Xinyu Zhao Zhihan Li Zhijie Zhang Pinpin Ji Qingyuan Liu Jiakai Zhao Jiahong Zhu Baoyuan Liu Xinjie Wang Yani Sun Qin Zhao |
| author_facet | Xu Chen Yueting Chang Lu Zhang Xinyu Zhao Zhihan Li Zhijie Zhang Pinpin Ji Qingyuan Liu Jiakai Zhao Jiahong Zhu Baoyuan Liu Xinjie Wang Yani Sun Qin Zhao |
| author_sort | Xu Chen |
| collection | DOAJ |
| description | Abstract Porcine reproductive and respiratory syndrome virus (PRRSV) infection causes significant economic loss to the global pig industry. Genotype 1 and 2 PRRSV (PRRSV-1 and -2) infections have been reported in China, Europe and America. For accurate prevention, nanobodies were first used as diagnostic reagents for PRRSV typing. In this study three nanobodies targeting both PRRSV-1 and -2, two targeting PRRSV-1 and three targeting PRRSV-2, were screened and produced. To develop two competitive ELISAs (cELISAs), the g1-2-PRRSV-Nb3-HRP nanobody was chosen for the g1-2-cELISA, to detect common antibodies against PRRSV-1 and -2, and the g1-PRRSV-Nb136-HRP nanobody was chosen for the g1-cELISA, to detect anti-PRRSV-1 antibodies. The two cELISAs were developed using PRRSV-1-N protein as coating antigen, and the amounts for both were 100 ng/well. The optimized dilution of testing pig sera was 1:20, the optimized reaction times were 30 min, and the colorimetric reaction times were 15 min. Then, the cut-off values of the g1-2-cELISA and g1-cELISA were 26.6% and 35.6%, respectively. Both of them have high sensitivity, strong specificity, good repeatability, and stability. In addition, for the 1534 clinical pig sera, an agreement rate of 99.02% (Kappa values = 0.97) was determined between the g1-2-cELISA and the commercial IDEXX ELISA kit. For the g1-cELSIA, it can specifically detect anti-PRRSV-1 antibodies in the clinical pig sera. Importantly, combining two nanobody-based cELISAs can differentially detect antibodies against PRRSV-1 and -2. Graphical abstract |
| first_indexed | 2024-04-24T16:19:49Z |
| format | Article |
| id | doaj.art-754610317da34670a12140bbb80b06b9 |
| institution | Directory Open Access Journal |
| issn | 2731-0442 |
| language | English |
| last_indexed | 2024-04-24T16:19:49Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Animal Diseases |
| spelling | doaj.art-754610317da34670a12140bbb80b06b92024-03-31T11:14:25ZengBMCAnimal Diseases2731-04422024-03-014111710.1186/s44149-024-00114-1Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virusXu Chen0Yueting Chang1Lu Zhang2Xinyu Zhao3Zhihan Li4Zhijie Zhang5Pinpin Ji6Qingyuan Liu7Jiakai Zhao8Jiahong Zhu9Baoyuan Liu10Xinjie Wang11Yani Sun12Qin Zhao13Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityShenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural SciencesDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F UniversityAbstract Porcine reproductive and respiratory syndrome virus (PRRSV) infection causes significant economic loss to the global pig industry. Genotype 1 and 2 PRRSV (PRRSV-1 and -2) infections have been reported in China, Europe and America. For accurate prevention, nanobodies were first used as diagnostic reagents for PRRSV typing. In this study three nanobodies targeting both PRRSV-1 and -2, two targeting PRRSV-1 and three targeting PRRSV-2, were screened and produced. To develop two competitive ELISAs (cELISAs), the g1-2-PRRSV-Nb3-HRP nanobody was chosen for the g1-2-cELISA, to detect common antibodies against PRRSV-1 and -2, and the g1-PRRSV-Nb136-HRP nanobody was chosen for the g1-cELISA, to detect anti-PRRSV-1 antibodies. The two cELISAs were developed using PRRSV-1-N protein as coating antigen, and the amounts for both were 100 ng/well. The optimized dilution of testing pig sera was 1:20, the optimized reaction times were 30 min, and the colorimetric reaction times were 15 min. Then, the cut-off values of the g1-2-cELISA and g1-cELISA were 26.6% and 35.6%, respectively. Both of them have high sensitivity, strong specificity, good repeatability, and stability. In addition, for the 1534 clinical pig sera, an agreement rate of 99.02% (Kappa values = 0.97) was determined between the g1-2-cELISA and the commercial IDEXX ELISA kit. For the g1-cELSIA, it can specifically detect anti-PRRSV-1 antibodies in the clinical pig sera. Importantly, combining two nanobody-based cELISAs can differentially detect antibodies against PRRSV-1 and -2. Graphical abstracthttps://doi.org/10.1186/s44149-024-00114-1PRRSVCompetitive ELISANanobodyAntigen epitope |
| spellingShingle | Xu Chen Yueting Chang Lu Zhang Xinyu Zhao Zhihan Li Zhijie Zhang Pinpin Ji Qingyuan Liu Jiakai Zhao Jiahong Zhu Baoyuan Liu Xinjie Wang Yani Sun Qin Zhao Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus Animal Diseases PRRSV Competitive ELISA Nanobody Antigen epitope |
| title | Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus |
| title_full | Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus |
| title_fullStr | Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus |
| title_full_unstemmed | Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus |
| title_short | Two nanobody-based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus |
| title_sort | two nanobody based immunoassays to differentiate antibodies against genotype 1 and 2 porcine reproductive and respiratory syndrome virus |
| topic | PRRSV Competitive ELISA Nanobody Antigen epitope |
| url | https://doi.org/10.1186/s44149-024-00114-1 |
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