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A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses

BACKGROUND: Chicken anemia virus (CAV), avian reovirus (ARV), infectious bursal disease virus (IBDV), Marek’s disease virus (MDV) and reticuloendotheliosis virus (REV) all cause immunosuppressive disease in birds through vertical or horizontal transmission. Mixed infections with these immunosuppress...

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Autores principales: Cong, Feng, Zhu, Yujun, Wang, Jing, Lian, Yuexiao, Liu, Xiangnan, Xiao, Li, Huang, Ren, Zhang, Yu, Chen, Meili, Guo, Pengju
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6238339/
https://www.ncbi.nlm.nih.gov/pubmed/30442149
http://dx.doi.org/10.1186/s12917-018-1663-1
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author Cong, Feng
Zhu, Yujun
Wang, Jing
Lian, Yuexiao
Liu, Xiangnan
Xiao, Li
Huang, Ren
Zhang, Yu
Chen, Meili
Guo, Pengju
author_facet Cong, Feng
Zhu, Yujun
Wang, Jing
Lian, Yuexiao
Liu, Xiangnan
Xiao, Li
Huang, Ren
Zhang, Yu
Chen, Meili
Guo, Pengju
author_sort Cong, Feng
collection PubMed
description BACKGROUND: Chicken anemia virus (CAV), avian reovirus (ARV), infectious bursal disease virus (IBDV), Marek’s disease virus (MDV) and reticuloendotheliosis virus (REV) all cause immunosuppressive disease in birds through vertical or horizontal transmission. Mixed infections with these immunosuppressive pathogens lead to atypical clinical signs and obstruct accurate diagnoses and epidemiological investigations. Therefore, it is essential to develop a high-throughput assay for the simultaneous detection of these immunosuppressive viruses with high specificity and sensitivity. The aim of this study was to establish a novel method using a RT-PCR assay combined with fluorescence labeled polystyrene bead microarray (multiplex xTAG assay) to detect single or mixed viral infections. RESULTS: The results showed that the established xTAG assay had no nonspecific reactions with avian influenza virus (AIV), infectious bronchitis virus (IBV), newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). The limit of detection was 1.0 × 10(3) copies/μL for IBDV and 1.0 × 10(2)copies/μL for the other four viruses. Ninety field samples were tested and the results were confirmed using conventional RT-PCR methods. The detection results of these two methods were 100% consistent. The established multiplex xTAG assay allows a high throughput and simultaneous detection of five chicken immunosuppressive viruses. CONCLUSION: The multiplex xTAG assay has been showed to be an additional tool for molecular epidemiology studies of five chicken immunosuppressive viruses in the poultry industry. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-018-1663-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-62383392018-11-26 A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses Cong, Feng Zhu, Yujun Wang, Jing Lian, Yuexiao Liu, Xiangnan Xiao, Li Huang, Ren Zhang, Yu Chen, Meili Guo, Pengju BMC Vet Res Research Article BACKGROUND: Chicken anemia virus (CAV), avian reovirus (ARV), infectious bursal disease virus (IBDV), Marek’s disease virus (MDV) and reticuloendotheliosis virus (REV) all cause immunosuppressive disease in birds through vertical or horizontal transmission. Mixed infections with these immunosuppressive pathogens lead to atypical clinical signs and obstruct accurate diagnoses and epidemiological investigations. Therefore, it is essential to develop a high-throughput assay for the simultaneous detection of these immunosuppressive viruses with high specificity and sensitivity. The aim of this study was to establish a novel method using a RT-PCR assay combined with fluorescence labeled polystyrene bead microarray (multiplex xTAG assay) to detect single or mixed viral infections. RESULTS: The results showed that the established xTAG assay had no nonspecific reactions with avian influenza virus (AIV), infectious bronchitis virus (IBV), newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). The limit of detection was 1.0 × 10(3) copies/μL for IBDV and 1.0 × 10(2)copies/μL for the other four viruses. Ninety field samples were tested and the results were confirmed using conventional RT-PCR methods. The detection results of these two methods were 100% consistent. The established multiplex xTAG assay allows a high throughput and simultaneous detection of five chicken immunosuppressive viruses. CONCLUSION: The multiplex xTAG assay has been showed to be an additional tool for molecular epidemiology studies of five chicken immunosuppressive viruses in the poultry industry. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-018-1663-1) contains supplementary material, which is available to authorized users. BioMed Central 2018-11-15 /pmc/articles/PMC6238339/ /pubmed/30442149 http://dx.doi.org/10.1186/s12917-018-1663-1 Text en © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Cong, Feng
Zhu, Yujun
Wang, Jing
Lian, Yuexiao
Liu, Xiangnan
Xiao, Li
Huang, Ren
Zhang, Yu
Chen, Meili
Guo, Pengju
A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses
title A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses
title_full A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses
title_fullStr A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses
title_full_unstemmed A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses
title_short A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses
title_sort multiplex xtag assay for the simultaneous detection of five chicken immunosuppressive viruses
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6238339/
https://www.ncbi.nlm.nih.gov/pubmed/30442149
http://dx.doi.org/10.1186/s12917-018-1663-1
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