Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus

Duck enteritis virus (DEV), goose parvovirus (GPV), and muscovy duck parvovirus (MDPV) all have similar symptoms after infection, such as severe diarrhea, which seriously affects the healthy development of the waterfowl industry. Hence, it is important to devise a rapid and precise assay for the det...

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Main Authors: Qian Qiu, Ruiming Hu, Zirui Liu, Linjie Yan, Fan Yang, Xueyan Dai, Chenghong Xing, Huabin Cao
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/15/11/1599
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author Qian Qiu
Ruiming Hu
Zirui Liu
Linjie Yan
Fan Yang
Xueyan Dai
Chenghong Xing
Huabin Cao
author_facet Qian Qiu
Ruiming Hu
Zirui Liu
Linjie Yan
Fan Yang
Xueyan Dai
Chenghong Xing
Huabin Cao
author_sort Qian Qiu
collection DOAJ
description Duck enteritis virus (DEV), goose parvovirus (GPV), and muscovy duck parvovirus (MDPV) all have similar symptoms after infection, such as severe diarrhea, which seriously affects the healthy development of the waterfowl industry. Hence, it is important to devise a rapid and precise assay for the detection of these three viruses. In this study, a TaqMan probe-based multi-quantitative polymerase chain reaction (qPCR) assay was developed and optimized. Three specific primers and probes were designed according to the conserved regions of UL6 of DEV, REP of GPV, and VP1 of MDPV, respectively. DEV demonstrated a detection limit of 11.6 copies, GPV detected a limit of 95 copies, and MDPV showcased a detection limit of 14.8 copies. The correlation coefficient is greater than 0.99, and the amplification efficiency is 89% to 93%. These results indicate that the multiplex qPCR assay has high sensitivity, specificity, and stability. Of the 215 clinical samples used in this study, 33 tested DEV positive, 25 tested GPV positive, and 24 tested MDPV positive. Overall, the assay established in the current study presents a rapid, efficient, specific, and sensitive tool for of detecting DEV, GPV, and MDPV.
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institution OA Journals
issn 2076-2615
language English
publishDate 2025-05-01
publisher MDPI AG
record_format Article
series Animals
spelling doaj-art-cf39eb45950d488c958eaf44e80408ed2025-08-20T02:22:56ZengMDPI AGAnimals2076-26152025-05-011511159910.3390/ani15111599Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck ParvovirusQian Qiu0Ruiming Hu1Zirui Liu2Linjie Yan3Fan Yang4Xueyan Dai5Chenghong Xing6Huabin Cao7Jiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaJiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, No. 22 Xinong Road, Yangling District, Xianyang 712000, ChinaJiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaJiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaJiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaJiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaJiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, ChinaDuck enteritis virus (DEV), goose parvovirus (GPV), and muscovy duck parvovirus (MDPV) all have similar symptoms after infection, such as severe diarrhea, which seriously affects the healthy development of the waterfowl industry. Hence, it is important to devise a rapid and precise assay for the detection of these three viruses. In this study, a TaqMan probe-based multi-quantitative polymerase chain reaction (qPCR) assay was developed and optimized. Three specific primers and probes were designed according to the conserved regions of UL6 of DEV, REP of GPV, and VP1 of MDPV, respectively. DEV demonstrated a detection limit of 11.6 copies, GPV detected a limit of 95 copies, and MDPV showcased a detection limit of 14.8 copies. The correlation coefficient is greater than 0.99, and the amplification efficiency is 89% to 93%. These results indicate that the multiplex qPCR assay has high sensitivity, specificity, and stability. Of the 215 clinical samples used in this study, 33 tested DEV positive, 25 tested GPV positive, and 24 tested MDPV positive. Overall, the assay established in the current study presents a rapid, efficient, specific, and sensitive tool for of detecting DEV, GPV, and MDPV.https://www.mdpi.com/2076-2615/15/11/1599duck enteritis virusgoose parvovirusmuscovy duck parvovirusmulti-quantitative qPCRTaqMan probe
spellingShingle Qian Qiu
Ruiming Hu
Zirui Liu
Linjie Yan
Fan Yang
Xueyan Dai
Chenghong Xing
Huabin Cao
Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus
Animals
duck enteritis virus
goose parvovirus
muscovy duck parvovirus
multi-quantitative qPCR
TaqMan probe
title Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus
title_full Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus
title_fullStr Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus
title_full_unstemmed Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus
title_short Development of a Multiplex Quantitative Polymerase Chain Reaction Assay for the Detection of Duck Enteritis Virus, Goose Parvovirus, and Muscovy Duck Parvovirus
title_sort development of a multiplex quantitative polymerase chain reaction assay for the detection of duck enteritis virus goose parvovirus and muscovy duck parvovirus
topic duck enteritis virus
goose parvovirus
muscovy duck parvovirus
multi-quantitative qPCR
TaqMan probe
url https://www.mdpi.com/2076-2615/15/11/1599
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