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Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda

BACKGROUND: Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially...

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Autores principales: Huang, Pei, Yu, Yue, Meng, Xianyong, Wang, Tiecheng, Yan, Feihu, Li, Entao, Shi, Zhikang, He, Hongbin, Yang, Songtao, Xia, Xianzhu, Wang, Jianzhong, Feng, Na
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8063184/
https://www.ncbi.nlm.nih.gov/pubmed/33892731
http://dx.doi.org/10.1186/s12917-021-02880-3
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author Huang, Pei
Yu, Yue
Meng, Xianyong
Wang, Tiecheng
Yan, Feihu
Li, Entao
Shi, Zhikang
He, Hongbin
Yang, Songtao
Xia, Xianzhu
Wang, Jianzhong
Feng, Na
author_facet Huang, Pei
Yu, Yue
Meng, Xianyong
Wang, Tiecheng
Yan, Feihu
Li, Entao
Shi, Zhikang
He, Hongbin
Yang, Songtao
Xia, Xianzhu
Wang, Jianzhong
Feng, Na
author_sort Huang, Pei
collection PubMed
description BACKGROUND: Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). RESULTS: The RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 10(1) copies/μl RNA transcripts and 10(0.5) TCID(50) ml(− 1) viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples. CONCLUSIONS: The RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda.
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spelling pubmed-80631842021-04-23 Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda Huang, Pei Yu, Yue Meng, Xianyong Wang, Tiecheng Yan, Feihu Li, Entao Shi, Zhikang He, Hongbin Yang, Songtao Xia, Xianzhu Wang, Jianzhong Feng, Na BMC Vet Res Research Article BACKGROUND: Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). RESULTS: The RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 10(1) copies/μl RNA transcripts and 10(0.5) TCID(50) ml(− 1) viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples. CONCLUSIONS: The RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda. BioMed Central 2021-04-23 /pmc/articles/PMC8063184/ /pubmed/33892731 http://dx.doi.org/10.1186/s12917-021-02880-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Huang, Pei
Yu, Yue
Meng, Xianyong
Wang, Tiecheng
Yan, Feihu
Li, Entao
Shi, Zhikang
He, Hongbin
Yang, Songtao
Xia, Xianzhu
Wang, Jianzhong
Feng, Na
Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
title Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
title_full Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
title_fullStr Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
title_full_unstemmed Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
title_short Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
title_sort development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8063184/
https://www.ncbi.nlm.nih.gov/pubmed/33892731
http://dx.doi.org/10.1186/s12917-021-02880-3
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