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Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein
Getah virus (GETV) disease is a mosquito-borne infectious disease that causes fever, aseptic meningitis, and abortion in a variety of animals. Currently, the epidemic trend of GETV disease increases seriously worldwide, especially in China, posing a potential threat to animal safety and public healt...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9685671/ https://www.ncbi.nlm.nih.gov/pubmed/36439788 http://dx.doi.org/10.3389/fmicb.2022.1029444 |
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author | Qiu, Xiangshu Cao, Xinyu Shi, Ning Zhang, He Zhu, Xiangyu Gao, Yan Mai, Zhanhai Jin, Ningyi Lu, Huijun |
author_facet | Qiu, Xiangshu Cao, Xinyu Shi, Ning Zhang, He Zhu, Xiangyu Gao, Yan Mai, Zhanhai Jin, Ningyi Lu, Huijun |
author_sort | Qiu, Xiangshu |
collection | PubMed |
description | Getah virus (GETV) disease is a mosquito-borne infectious disease that causes fever, aseptic meningitis, and abortion in a variety of animals. Currently, the epidemic trend of GETV disease increases seriously worldwide, especially in China, posing a potential threat to animal safety and public health. However, there are few reports about the epidemiological investigation of GETV disease in China as well as a lack of commercial diagnostic kit for GETV antibody. Therefore, the establishment of a rapid, sensitive and suitable GETV antibody detection method for large-scale samples is an urgent request to fully understand the prevalence of GETV disease. Here, a recombinant plasmid pET22b-GETV-E2d that contained the domain of GETV-E2 (E2d) fused to His-tag was constructed to express recombinant protein E2d (rE2d) in Escherichia coli. The rE2d was mainly expressed in inclusion bodies. And it was purified successfully by nickel affinity column so that it could be used to develop an indirect ELISA (rE2d-ELISA). After optimizing reaction conditions of rE2d-ELISA, the cut-off value was determined as 0.396 with 100 equine sera tested by virus neutralization test (VNT). Furthermore, rE2d-ELISA method showed the positive rate of IgG antibodies against GETV was 54.3% based on testing 646 clinical serum samples obtained in Xinjiang whereas the overall coincidence rate between rE2d-ELISA and VNT was 94.0%, with 98.2% sensitivity and 92.6% specificity. The findings suggest that the developed IgG ELISA employing recombinant E2d promises was an efficient and low-cost type of antibody detection method for horse, which will benefit for prevention of GETV outbreaks in horses. |
format | Online Article Text |
id | pubmed-9685671 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-96856712022-11-25 Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein Qiu, Xiangshu Cao, Xinyu Shi, Ning Zhang, He Zhu, Xiangyu Gao, Yan Mai, Zhanhai Jin, Ningyi Lu, Huijun Front Microbiol Microbiology Getah virus (GETV) disease is a mosquito-borne infectious disease that causes fever, aseptic meningitis, and abortion in a variety of animals. Currently, the epidemic trend of GETV disease increases seriously worldwide, especially in China, posing a potential threat to animal safety and public health. However, there are few reports about the epidemiological investigation of GETV disease in China as well as a lack of commercial diagnostic kit for GETV antibody. Therefore, the establishment of a rapid, sensitive and suitable GETV antibody detection method for large-scale samples is an urgent request to fully understand the prevalence of GETV disease. Here, a recombinant plasmid pET22b-GETV-E2d that contained the domain of GETV-E2 (E2d) fused to His-tag was constructed to express recombinant protein E2d (rE2d) in Escherichia coli. The rE2d was mainly expressed in inclusion bodies. And it was purified successfully by nickel affinity column so that it could be used to develop an indirect ELISA (rE2d-ELISA). After optimizing reaction conditions of rE2d-ELISA, the cut-off value was determined as 0.396 with 100 equine sera tested by virus neutralization test (VNT). Furthermore, rE2d-ELISA method showed the positive rate of IgG antibodies against GETV was 54.3% based on testing 646 clinical serum samples obtained in Xinjiang whereas the overall coincidence rate between rE2d-ELISA and VNT was 94.0%, with 98.2% sensitivity and 92.6% specificity. The findings suggest that the developed IgG ELISA employing recombinant E2d promises was an efficient and low-cost type of antibody detection method for horse, which will benefit for prevention of GETV outbreaks in horses. Frontiers Media S.A. 2022-11-10 /pmc/articles/PMC9685671/ /pubmed/36439788 http://dx.doi.org/10.3389/fmicb.2022.1029444 Text en Copyright © 2022 Qiu, Cao, Shi, Zhang, Zhu, Gao, Mai, Jin and Lu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Qiu, Xiangshu Cao, Xinyu Shi, Ning Zhang, He Zhu, Xiangyu Gao, Yan Mai, Zhanhai Jin, Ningyi Lu, Huijun Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein |
title | Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein |
title_full | Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein |
title_fullStr | Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein |
title_full_unstemmed | Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein |
title_short | Development and application of an indirect ELISA for detecting equine IgG antibodies against Getah virus with recombinant E2 domain protein |
title_sort | development and application of an indirect elisa for detecting equine igg antibodies against getah virus with recombinant e2 domain protein |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9685671/ https://www.ncbi.nlm.nih.gov/pubmed/36439788 http://dx.doi.org/10.3389/fmicb.2022.1029444 |
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