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A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies

Pseudorabies caused by pseudorabies virus (PRV) infection is still a major disease affecting the pig industry; its eradication depends on effective vaccination and antibody (Ab) detection. For a more rapid and accurate PRV detection method that is suitable for clinical application, here, we establis...

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Autores principales: Pan, Jiaojiao, Li, Yufang, Wang, Tongyan, Chang, Jingfeng, Hao, Liying, Chen, Junjie, Peng, Wuping, Deng, Junhua, Huang, Baicheng, Tian, Kegong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9360482/
https://www.ncbi.nlm.nih.gov/pubmed/35959367
http://dx.doi.org/10.3389/fcimb.2022.912108
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author Pan, Jiaojiao
Li, Yufang
Wang, Tongyan
Chang, Jingfeng
Hao, Liying
Chen, Junjie
Peng, Wuping
Deng, Junhua
Huang, Baicheng
Tian, Kegong
author_facet Pan, Jiaojiao
Li, Yufang
Wang, Tongyan
Chang, Jingfeng
Hao, Liying
Chen, Junjie
Peng, Wuping
Deng, Junhua
Huang, Baicheng
Tian, Kegong
author_sort Pan, Jiaojiao
collection PubMed
description Pseudorabies caused by pseudorabies virus (PRV) infection is still a major disease affecting the pig industry; its eradication depends on effective vaccination and antibody (Ab) detection. For a more rapid and accurate PRV detection method that is suitable for clinical application, here, we established a poly(dimethylsiloxane)-based (efficient removal of non-specific binding) solid-phase protein chip platform (blocking ELISA) for dual detection of PRV gD and gE Abs. The purified gD and gE proteins expressed in baculovirus were coated into the highly hydrophobic nanomembrane by an automatic spotter, and the gray values measured by a scanner were used for the S/N (sample/negative) value calculation (gD and gE Abs standard, positive: S/N value ≤0.6; negative: S/N value >0.7; suspicious: 0.6 < S/N ≤ 0.7). The method showed an equal sensitivity in the gD Ab test of immunized pig serum samples compared to the neutralization test and higher sensitivity in the gE Ab test compared to the commercial gE Ab detection kit. In the clinical evaluation, we found an agreement of 100% (122/122) in the gD Ab detection compared to the neutralization test and an agreement of 97.5% (119/122) in the gE Ab detection compared to the commercial PRV gE Ab detection kit. In summary, the protein chip platform for dual detection of PRV gD and gE Abs showed high sensitivity and specificity, which is suitable for PRV immune efficacy evaluation and epidemic monitoring.
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spelling pubmed-93604822022-08-10 A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies Pan, Jiaojiao Li, Yufang Wang, Tongyan Chang, Jingfeng Hao, Liying Chen, Junjie Peng, Wuping Deng, Junhua Huang, Baicheng Tian, Kegong Front Cell Infect Microbiol Cellular and Infection Microbiology Pseudorabies caused by pseudorabies virus (PRV) infection is still a major disease affecting the pig industry; its eradication depends on effective vaccination and antibody (Ab) detection. For a more rapid and accurate PRV detection method that is suitable for clinical application, here, we established a poly(dimethylsiloxane)-based (efficient removal of non-specific binding) solid-phase protein chip platform (blocking ELISA) for dual detection of PRV gD and gE Abs. The purified gD and gE proteins expressed in baculovirus were coated into the highly hydrophobic nanomembrane by an automatic spotter, and the gray values measured by a scanner were used for the S/N (sample/negative) value calculation (gD and gE Abs standard, positive: S/N value ≤0.6; negative: S/N value >0.7; suspicious: 0.6 < S/N ≤ 0.7). The method showed an equal sensitivity in the gD Ab test of immunized pig serum samples compared to the neutralization test and higher sensitivity in the gE Ab test compared to the commercial gE Ab detection kit. In the clinical evaluation, we found an agreement of 100% (122/122) in the gD Ab detection compared to the neutralization test and an agreement of 97.5% (119/122) in the gE Ab detection compared to the commercial PRV gE Ab detection kit. In summary, the protein chip platform for dual detection of PRV gD and gE Abs showed high sensitivity and specificity, which is suitable for PRV immune efficacy evaluation and epidemic monitoring. Frontiers Media S.A. 2022-07-26 /pmc/articles/PMC9360482/ /pubmed/35959367 http://dx.doi.org/10.3389/fcimb.2022.912108 Text en Copyright © 2022 Pan, Li, Wang, Chang, Hao, Chen, Peng, Deng, Huang and Tian 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 Cellular and Infection Microbiology
Pan, Jiaojiao
Li, Yufang
Wang, Tongyan
Chang, Jingfeng
Hao, Liying
Chen, Junjie
Peng, Wuping
Deng, Junhua
Huang, Baicheng
Tian, Kegong
A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies
title A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies
title_full A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies
title_fullStr A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies
title_full_unstemmed A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies
title_short A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies
title_sort poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of pseudorabies virus gd and ge antibodies
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9360482/
https://www.ncbi.nlm.nih.gov/pubmed/35959367
http://dx.doi.org/10.3389/fcimb.2022.912108
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