Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus
Equine Infectious Anemia Virus (EIAV) poses significant diagnostic challenges due to its genetic variability and the limitations of conventional nucleic acid detection methods. This study developed an antigen-capture, enzyme-linked immunosorbent assay (AC-ELISA) for the detection and quantification...
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MDPI AG
2025-06-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/13/7/1500 |
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| author | Haibing Liang Bingqian Zhou Zhe Hu Xiaoyu Chu Xuefeng Wang Cheng Du Xiaojun Wang |
| author_facet | Haibing Liang Bingqian Zhou Zhe Hu Xiaoyu Chu Xuefeng Wang Cheng Du Xiaojun Wang |
| author_sort | Haibing Liang |
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| description | Equine Infectious Anemia Virus (EIAV) poses significant diagnostic challenges due to its genetic variability and the limitations of conventional nucleic acid detection methods. This study developed an antigen-capture, enzyme-linked immunosorbent assay (AC-ELISA) for the detection and quantification of the EIAV capsid protein p26. The assay utilized a monoclonal antibody (1G11) specific to the p26 protein as the capture antibody and a polyclonal antibody as the detection antibody, forming a highly specific and sensitive detection system. Under optimized conditions, the detection limit of the AC-ELISA was 1.95 ng/mL, with a good linear relationship observed between 1.95 ng/mL and 60.5 ng/mL of p26 protein. Additionally, the AC-ELISA effectively distinguished EIAV from other equine viruses, including equine herpesvirus 1 (EHV-1), equine arteritis virus (EAV), and equine influenza virus (EIV), without cross-reactivity. Importantly, the AC-ELISA demonstrated the ability to detect multiple EIAV strains, including virulent strains, attenuated strains, and strains from other countries, highlighting its broad applicability across diverse EIAV isolates. Compared to western blot and reverse transcriptase assays, the AC-ELISA exhibited higher sensitivity and strong correlation in quantifying the EIAV p26 protein. The assay is simple, rapid, and cost-effective, making it suitable for both laboratory research and clinical applications. It provides a powerful tool for EIAV detection and quantification, supporting future vaccine development and clinical trials. |
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| institution | Kabale University |
| issn | 2076-2607 |
| language | English |
| publishDate | 2025-06-01 |
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| series | Microorganisms |
| spelling | doaj-art-23bfd851714a4eaf8ad4fd2b69f324032025-08-20T03:56:46ZengMDPI AGMicroorganisms2076-26072025-06-01137150010.3390/microorganisms13071500Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia VirusHaibing Liang0Bingqian Zhou1Zhe Hu2Xiaoyu Chu3Xuefeng Wang4Cheng Du5Xiaojun Wang6State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, ChinaEquine Infectious Anemia Virus (EIAV) poses significant diagnostic challenges due to its genetic variability and the limitations of conventional nucleic acid detection methods. This study developed an antigen-capture, enzyme-linked immunosorbent assay (AC-ELISA) for the detection and quantification of the EIAV capsid protein p26. The assay utilized a monoclonal antibody (1G11) specific to the p26 protein as the capture antibody and a polyclonal antibody as the detection antibody, forming a highly specific and sensitive detection system. Under optimized conditions, the detection limit of the AC-ELISA was 1.95 ng/mL, with a good linear relationship observed between 1.95 ng/mL and 60.5 ng/mL of p26 protein. Additionally, the AC-ELISA effectively distinguished EIAV from other equine viruses, including equine herpesvirus 1 (EHV-1), equine arteritis virus (EAV), and equine influenza virus (EIV), without cross-reactivity. Importantly, the AC-ELISA demonstrated the ability to detect multiple EIAV strains, including virulent strains, attenuated strains, and strains from other countries, highlighting its broad applicability across diverse EIAV isolates. Compared to western blot and reverse transcriptase assays, the AC-ELISA exhibited higher sensitivity and strong correlation in quantifying the EIAV p26 protein. The assay is simple, rapid, and cost-effective, making it suitable for both laboratory research and clinical applications. It provides a powerful tool for EIAV detection and quantification, supporting future vaccine development and clinical trials.https://www.mdpi.com/2076-2607/13/7/1500EIAVAC-ELISAp26 |
| spellingShingle | Haibing Liang Bingqian Zhou Zhe Hu Xiaoyu Chu Xuefeng Wang Cheng Du Xiaojun Wang Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus Microorganisms EIAV AC-ELISA p26 |
| title | Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus |
| title_full | Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus |
| title_fullStr | Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus |
| title_full_unstemmed | Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus |
| title_short | Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus |
| title_sort | development of a broad spectrum antigen capture elisa using combined anti p26 polyclonal and monoclonal antibodies for detection of equine infectious anemia virus |
| topic | EIAV AC-ELISA p26 |
| url | https://www.mdpi.com/2076-2607/13/7/1500 |
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