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Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay
BACKGROUND: Domestic rabbits especially New Zealand white rabbits play an important role in biological research. The disease surveillance and quality control are essential to guarantee the results of animal experiments performed on rabbits. Rabbit hemorrhagic disease virus, rabbit rotavirus and Send...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5889542/ https://www.ncbi.nlm.nih.gov/pubmed/29625588 http://dx.doi.org/10.1186/s12917-018-1438-8 |
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author | Wu, Miaoli Zhu, Yujun Cong, Feng Rao, Dan Yuan, Wen Wang, Jing Huang, Bihong Lian, Yuexiao Zhang, Yu Huang, Ren Guo, Pengju |
author_facet | Wu, Miaoli Zhu, Yujun Cong, Feng Rao, Dan Yuan, Wen Wang, Jing Huang, Bihong Lian, Yuexiao Zhang, Yu Huang, Ren Guo, Pengju |
author_sort | Wu, Miaoli |
collection | PubMed |
description | BACKGROUND: Domestic rabbits especially New Zealand white rabbits play an important role in biological research. The disease surveillance and quality control are essential to guarantee the results of animal experiments performed on rabbits. Rabbit hemorrhagic disease virus, rabbit rotavirus and Sendai virus are the important pathogens that needed to be eliminated. Rapid and sensitive method focus on these three viruses should be established for routine monitoring. The Luminex x-TAG assay based on multiplex PCR and fluorescent microsphere is a fast developing technology applied in high throughput detection. Specific primers modified with oligonucleotide sequence/biotin were used to amplify target fragments. The conjugation between oligonucleotide sequence of the PCR products and the MagPlex-TAG microspheres was specific without any cross-reaction, and the hybridization products could be analyzed using the Luminex 200 analyzer instrument. Recombinant plasmids were constructed to estimate the detection limit of the viruses. Furthermore, 40 clinical samples were used to evaluate the efficiency of this multiplex PCR based Luminex x-TAG assay. RESULTS: According to the results, this new method showed high specificity and good stability. Assessed by the recombinant plasmids, the detection limit of three viruses was 100copies/μl. Among 40 clinical specimens, 18 specimens were found positive, which was completely concordant with the conventional PCR method. CONCLUSIONS: The new developed Luminex x-TAG assay is an accurate, high throughput method for rapid detection of three important viruses of rabbits. |
format | Online Article Text |
id | pubmed-5889542 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-58895422018-04-10 Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay Wu, Miaoli Zhu, Yujun Cong, Feng Rao, Dan Yuan, Wen Wang, Jing Huang, Bihong Lian, Yuexiao Zhang, Yu Huang, Ren Guo, Pengju BMC Vet Res Methodology Article BACKGROUND: Domestic rabbits especially New Zealand white rabbits play an important role in biological research. The disease surveillance and quality control are essential to guarantee the results of animal experiments performed on rabbits. Rabbit hemorrhagic disease virus, rabbit rotavirus and Sendai virus are the important pathogens that needed to be eliminated. Rapid and sensitive method focus on these three viruses should be established for routine monitoring. The Luminex x-TAG assay based on multiplex PCR and fluorescent microsphere is a fast developing technology applied in high throughput detection. Specific primers modified with oligonucleotide sequence/biotin were used to amplify target fragments. The conjugation between oligonucleotide sequence of the PCR products and the MagPlex-TAG microspheres was specific without any cross-reaction, and the hybridization products could be analyzed using the Luminex 200 analyzer instrument. Recombinant plasmids were constructed to estimate the detection limit of the viruses. Furthermore, 40 clinical samples were used to evaluate the efficiency of this multiplex PCR based Luminex x-TAG assay. RESULTS: According to the results, this new method showed high specificity and good stability. Assessed by the recombinant plasmids, the detection limit of three viruses was 100copies/μl. Among 40 clinical specimens, 18 specimens were found positive, which was completely concordant with the conventional PCR method. CONCLUSIONS: The new developed Luminex x-TAG assay is an accurate, high throughput method for rapid detection of three important viruses of rabbits. BioMed Central 2018-04-07 /pmc/articles/PMC5889542/ /pubmed/29625588 http://dx.doi.org/10.1186/s12917-018-1438-8 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 | Methodology Article Wu, Miaoli Zhu, Yujun Cong, Feng Rao, Dan Yuan, Wen Wang, Jing Huang, Bihong Lian, Yuexiao Zhang, Yu Huang, Ren Guo, Pengju Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay |
title | Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay |
title_full | Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay |
title_fullStr | Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay |
title_full_unstemmed | Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay |
title_short | Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay |
title_sort | rapid detection of three rabbit pathogens by use of the luminex x-tag assay |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5889542/ https://www.ncbi.nlm.nih.gov/pubmed/29625588 http://dx.doi.org/10.1186/s12917-018-1438-8 |
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