Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus
Bovine rhinitis B virus (BRBV) has been frequently identified in cattle diagnosed with bovine respiratory disease complex (BRDC) in recent years, suggesting its potential contribution to BRDC. The goal of this study was to develop a TaqMan-based real-time quantitative RT-PCR assay for efficient BRBV...
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Format: | Article |
Language: | English |
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Frontiers Media S.A.
2021-06-01
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Series: | Frontiers in Veterinary Science |
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Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2021.680707/full |
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doaj-401edf128d6b48f6a27636ebb5ce8df8 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yi-Lun Xie Yi-Lun Xie Dian-Hong Lv Xiao-Hui Wen Qi Zhai Man-Lin Luo Wen-Kang Wei Wen-Kang Wei Qin-Ling Chen Qin-Ling Chen Shao-Lun Zhai |
spellingShingle |
Yi-Lun Xie Yi-Lun Xie Dian-Hong Lv Xiao-Hui Wen Qi Zhai Man-Lin Luo Wen-Kang Wei Wen-Kang Wei Qin-Ling Chen Qin-Ling Chen Shao-Lun Zhai Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus Frontiers in Veterinary Science bovine rhinitis B virus RT-qPCR TaqMan probe detection assay |
author_facet |
Yi-Lun Xie Yi-Lun Xie Dian-Hong Lv Xiao-Hui Wen Qi Zhai Man-Lin Luo Wen-Kang Wei Wen-Kang Wei Qin-Ling Chen Qin-Ling Chen Shao-Lun Zhai |
author_sort |
Yi-Lun Xie |
title |
Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus |
title_short |
Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus |
title_full |
Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus |
title_fullStr |
Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus |
title_full_unstemmed |
Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B Virus |
title_sort |
development of a real-time quantitative rt-pcr assay for detection of bovine rhinitis b virus |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Veterinary Science |
issn |
2297-1769 |
publishDate |
2021-06-01 |
description |
Bovine rhinitis B virus (BRBV) has been frequently identified in cattle diagnosed with bovine respiratory disease complex (BRDC) in recent years, suggesting its potential contribution to BRDC. The goal of this study was to develop a TaqMan-based real-time quantitative RT-PCR assay for efficient BRBV detection. A pair of primers and a probe were designed based on the 3D gene of the BRBV genome. The assay was specific for BRBV and able to exclude bovine rhinitis A virus, foot-and-mouth disease virus and Senecavirus A. The limit of detection of the assay was 4.46 copies per reaction. A standard curve was plotted, with a coefficient of determination of 0.999 in the concentration range of 100-108 copies/μl. The reproducibility of the assay was acceptable, with the standard deviations of cycle threshold values lower than 1.00 in both intra- and inter-assay. Of 200 samples collected from 150 head of cattle in recent years in China, 11% (22/200) of the samples tested positive in the assay, i.e., 4.6% (7/150) of the cattle were BRBV positive. This study provides an efficient diagnostic tool for the epidemiological investigations of BRBV. |
topic |
bovine rhinitis B virus RT-qPCR TaqMan probe detection assay |
url |
https://www.frontiersin.org/articles/10.3389/fvets.2021.680707/full |
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spelling |
doaj-401edf128d6b48f6a27636ebb5ce8df82021-06-04T06:13:19ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692021-06-01810.3389/fvets.2021.680707680707Development of a Real-Time Quantitative RT-PCR Assay for Detection of Bovine Rhinitis B VirusYi-Lun Xie0Yi-Lun Xie1Dian-Hong Lv2Xiao-Hui Wen3Qi Zhai4Man-Lin Luo5Wen-Kang Wei6Wen-Kang Wei7Qin-Ling Chen8Qin-Ling Chen9Shao-Lun Zhai10College of Veterinary Medicine, South China Agricultural University, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaAgro-biological Gene Research Center, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaAgro-biological Gene Research Center, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaScientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Key Laboratory of Animal Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, ChinaBovine rhinitis B virus (BRBV) has been frequently identified in cattle diagnosed with bovine respiratory disease complex (BRDC) in recent years, suggesting its potential contribution to BRDC. The goal of this study was to develop a TaqMan-based real-time quantitative RT-PCR assay for efficient BRBV detection. A pair of primers and a probe were designed based on the 3D gene of the BRBV genome. The assay was specific for BRBV and able to exclude bovine rhinitis A virus, foot-and-mouth disease virus and Senecavirus A. The limit of detection of the assay was 4.46 copies per reaction. A standard curve was plotted, with a coefficient of determination of 0.999 in the concentration range of 100-108 copies/μl. The reproducibility of the assay was acceptable, with the standard deviations of cycle threshold values lower than 1.00 in both intra- and inter-assay. Of 200 samples collected from 150 head of cattle in recent years in China, 11% (22/200) of the samples tested positive in the assay, i.e., 4.6% (7/150) of the cattle were BRBV positive. This study provides an efficient diagnostic tool for the epidemiological investigations of BRBV.https://www.frontiersin.org/articles/10.3389/fvets.2021.680707/fullbovine rhinitis B virusRT-qPCRTaqMan probedetectionassay |