Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay
Abstract 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...
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| Format: | Article |
| Language: | English |
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BMC
2018-04-01
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| Series: | BMC Veterinary Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12917-018-1438-8 |
| _version_ | 1818545548772245504 |
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| author | Miaoli Wu Yujun Zhu Feng Cong Dan Rao Wen Yuan Jing Wang Bihong Huang Yuexiao Lian Yu Zhang Ren Huang Pengju Guo |
| author_facet | Miaoli Wu Yujun Zhu Feng Cong Dan Rao Wen Yuan Jing Wang Bihong Huang Yuexiao Lian Yu Zhang Ren Huang Pengju Guo |
| author_sort | Miaoli Wu |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-12T07:41:35Z |
| format | Article |
| id | doaj.art-b4ee5d23ee2344a080a67640b10b1e2f |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-12T07:41:35Z |
| publishDate | 2018-04-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-b4ee5d23ee2344a080a67640b10b1e2f2022-12-22T00:32:47ZengBMCBMC Veterinary Research1746-61482018-04-011411610.1186/s12917-018-1438-8Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assayMiaoli Wu0Yujun Zhu1Feng Cong2Dan Rao3Wen Yuan4Jing Wang5Bihong Huang6Yuexiao Lian7Yu Zhang8Ren Huang9Pengju Guo10Guangdong Provincial Key Laboratory of Laboratory AnimalsGuangdong Laboratory Animal Monitoring InstituteGuangdong Provincial Key Laboratory of Laboratory AnimalsGuangdong Laboratory Animal Monitoring InstituteGuangdong Provincial Key Laboratory of Laboratory AnimalsGuangdong Provincial Key Laboratory of Laboratory AnimalsGuangdong Laboratory Animal Monitoring InstituteGuangdong Laboratory Animal Monitoring InstituteGuangdong Laboratory Animal Monitoring InstituteGuangdong laboratory animals monitoring instituteGuangdong key laboratory of laboratory AnimalsAbstract 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.http://link.springer.com/article/10.1186/s12917-018-1438-8Rabbit hemorrhagic disease virusRabbit rotavirusSendai virusMultiple PCRLuminex x-TAG |
| spellingShingle | Miaoli Wu Yujun Zhu Feng Cong Dan Rao Wen Yuan Jing Wang Bihong Huang Yuexiao Lian Yu Zhang Ren Huang Pengju Guo Rapid detection of three rabbit pathogens by use of the Luminex x-TAG assay BMC Veterinary Research Rabbit hemorrhagic disease virus Rabbit rotavirus Sendai virus Multiple PCR Luminex x-TAG |
| 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 | Rabbit hemorrhagic disease virus Rabbit rotavirus Sendai virus Multiple PCR Luminex x-TAG |
| url | http://link.springer.com/article/10.1186/s12917-018-1438-8 |
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