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Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay

BACKGROUND: Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects. Specific MDPV detection is often rife with false positive results because of high identity at the genomic nucleotide level and antigenic...

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Autores principales: Wan, Chunhe, Chen, Cuiteng, Cheng, Longfei, Chen, Hongmei, Fu, Qiuling, Shi, Shaohua, Fu, Guanghua, Liu, Rongchang, Huang, Yu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6122767/
https://www.ncbi.nlm.nih.gov/pubmed/30176903
http://dx.doi.org/10.1186/s12917-018-1600-3
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author Wan, Chunhe
Chen, Cuiteng
Cheng, Longfei
Chen, Hongmei
Fu, Qiuling
Shi, Shaohua
Fu, Guanghua
Liu, Rongchang
Huang, Yu
author_facet Wan, Chunhe
Chen, Cuiteng
Cheng, Longfei
Chen, Hongmei
Fu, Qiuling
Shi, Shaohua
Fu, Guanghua
Liu, Rongchang
Huang, Yu
author_sort Wan, Chunhe
collection PubMed
description BACKGROUND: Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects. Specific MDPV detection is often rife with false positive results because of high identity at the genomic nucleotide level and antigenic similarity with goose parvovirus (GPV). The objective of this study was to develop a sensitive, highly specific, and repeatable TaqMan-based real-time PCR (qPCR) assay for facilitating the molecular detection of MDPV. RESULTS: The specific primers and probe were designed based on the conserved regions within MDPVs, but there was a variation in GPVs of the nonstructural (NS) genes after genetic comparison. After the optimization of qPCR conditions, the detection limit of this qPCR assay was 29.7 copies/μl. The assay was highly specific for the detection of MDPV, and no cross-reactivity was observed with other non-targeted duck-derived pathogens. Intra- and inter-assay variability was less than 2.21%, means a high degree of repeatability. The diagnostic applicability of the qPCR assay was proven that MDPV-positive can be found in cloacal swabs samples, Muscovy duck embryos and newly hatched Muscovy ducklings. CONCLUSIONS: Our data provided incidents that MDPV could be possible vertically transmitted from breeder Muscovy ducks to Muscovy ducklings. The developed qPCR assay in the study could be a reliable and specific tool for epidemiological surveillance and pathogenesis studies of MDPV.
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spelling pubmed-61227672018-09-10 Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay Wan, Chunhe Chen, Cuiteng Cheng, Longfei Chen, Hongmei Fu, Qiuling Shi, Shaohua Fu, Guanghua Liu, Rongchang Huang, Yu BMC Vet Res Research Article BACKGROUND: Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects. Specific MDPV detection is often rife with false positive results because of high identity at the genomic nucleotide level and antigenic similarity with goose parvovirus (GPV). The objective of this study was to develop a sensitive, highly specific, and repeatable TaqMan-based real-time PCR (qPCR) assay for facilitating the molecular detection of MDPV. RESULTS: The specific primers and probe were designed based on the conserved regions within MDPVs, but there was a variation in GPVs of the nonstructural (NS) genes after genetic comparison. After the optimization of qPCR conditions, the detection limit of this qPCR assay was 29.7 copies/μl. The assay was highly specific for the detection of MDPV, and no cross-reactivity was observed with other non-targeted duck-derived pathogens. Intra- and inter-assay variability was less than 2.21%, means a high degree of repeatability. The diagnostic applicability of the qPCR assay was proven that MDPV-positive can be found in cloacal swabs samples, Muscovy duck embryos and newly hatched Muscovy ducklings. CONCLUSIONS: Our data provided incidents that MDPV could be possible vertically transmitted from breeder Muscovy ducks to Muscovy ducklings. The developed qPCR assay in the study could be a reliable and specific tool for epidemiological surveillance and pathogenesis studies of MDPV. BioMed Central 2018-09-03 /pmc/articles/PMC6122767/ /pubmed/30176903 http://dx.doi.org/10.1186/s12917-018-1600-3 Text en © The Author(s). 2018 Open AccessThis 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
Wan, Chunhe
Chen, Cuiteng
Cheng, Longfei
Chen, Hongmei
Fu, Qiuling
Shi, Shaohua
Fu, Guanghua
Liu, Rongchang
Huang, Yu
Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay
title Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay
title_full Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay
title_fullStr Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay
title_full_unstemmed Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay
title_short Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay
title_sort specific detection of muscovy duck parvovirus infection by taqman-based real-time pcr assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6122767/
https://www.ncbi.nlm.nih.gov/pubmed/30176903
http://dx.doi.org/10.1186/s12917-018-1600-3
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