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Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction

BACKGROUND: Peste des petits ruminants (PPR) disease is a cross-species infectious disease that severely affects small ruminants and causes great losses to livestock industries in various countries. Distinguishing vaccine-immunized animals from naturally infected animals is an important prerequisite...

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Autores principales: Tang, Jingyu, Du, Hanyu, Tang, Aoxing, Jia, Nannan, Zhu, Jie, Li, Chuanfeng, Meng, Chunchun, Liu, Guangqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9841696/
https://www.ncbi.nlm.nih.gov/pubmed/36647038
http://dx.doi.org/10.1186/s12917-023-03568-6
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author Tang, Jingyu
Du, Hanyu
Tang, Aoxing
Jia, Nannan
Zhu, Jie
Li, Chuanfeng
Meng, Chunchun
Liu, Guangqing
author_facet Tang, Jingyu
Du, Hanyu
Tang, Aoxing
Jia, Nannan
Zhu, Jie
Li, Chuanfeng
Meng, Chunchun
Liu, Guangqing
author_sort Tang, Jingyu
collection PubMed
description BACKGROUND: Peste des petits ruminants (PPR) disease is a cross-species infectious disease that severely affects small ruminants and causes great losses to livestock industries in various countries. Distinguishing vaccine-immunized animals from naturally infected animals is an important prerequisite for the eradication of PPR. At present PPRV are classified into lineages I through IV, and only one vaccination strain, Nigeria/75/1, belongs to lineage II, but all of the epidemic strains in China at present are from lineage IV. RESULTS: To achieve this goal, we developed an SYBR Green I real-time qRT-PCR method for rapid detection and identification of PPRV lineages II and IV by analyzing different melting curve analyses. The negative amplification of other commonly circulating viruses such as orf virus, goat poxvirus, and foot-and-mouth disease virus demonstrated that primers targeting the L gene of PPRV were extremely specific. The sensitivity of the assay was assessed based on plasmid DNA and the detection limit achieved was 100 copies of PPRV lineages II and IV. CONCLUSION: Since the method has high sensitivity, specificity, and reproducibility, it will be effectively differentiated PPRV lineages II from PPRV lineages IV in PPRV infected animals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-023-03568-6.
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spelling pubmed-98416962023-01-17 Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction Tang, Jingyu Du, Hanyu Tang, Aoxing Jia, Nannan Zhu, Jie Li, Chuanfeng Meng, Chunchun Liu, Guangqing BMC Vet Res Research BACKGROUND: Peste des petits ruminants (PPR) disease is a cross-species infectious disease that severely affects small ruminants and causes great losses to livestock industries in various countries. Distinguishing vaccine-immunized animals from naturally infected animals is an important prerequisite for the eradication of PPR. At present PPRV are classified into lineages I through IV, and only one vaccination strain, Nigeria/75/1, belongs to lineage II, but all of the epidemic strains in China at present are from lineage IV. RESULTS: To achieve this goal, we developed an SYBR Green I real-time qRT-PCR method for rapid detection and identification of PPRV lineages II and IV by analyzing different melting curve analyses. The negative amplification of other commonly circulating viruses such as orf virus, goat poxvirus, and foot-and-mouth disease virus demonstrated that primers targeting the L gene of PPRV were extremely specific. The sensitivity of the assay was assessed based on plasmid DNA and the detection limit achieved was 100 copies of PPRV lineages II and IV. CONCLUSION: Since the method has high sensitivity, specificity, and reproducibility, it will be effectively differentiated PPRV lineages II from PPRV lineages IV in PPRV infected animals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-023-03568-6. BioMed Central 2023-01-16 /pmc/articles/PMC9841696/ /pubmed/36647038 http://dx.doi.org/10.1186/s12917-023-03568-6 Text en © The Author(s) 2023 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
Tang, Jingyu
Du, Hanyu
Tang, Aoxing
Jia, Nannan
Zhu, Jie
Li, Chuanfeng
Meng, Chunchun
Liu, Guangqing
Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction
title Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction
title_full Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction
title_fullStr Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction
title_full_unstemmed Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction
title_short Simultaneous detection and identification of Peste des petits ruminants Virus Lineages II and IV by MCA-Based real-time quantitative RT-PCR assay within single reaction
title_sort simultaneous detection and identification of peste des petits ruminants virus lineages ii and iv by mca-based real-time quantitative rt-pcr assay within single reaction
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9841696/
https://www.ncbi.nlm.nih.gov/pubmed/36647038
http://dx.doi.org/10.1186/s12917-023-03568-6
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