Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus
Rift Valley fever (RVF) is a severe infectious disease, which can through mosquito bites, direct contact and aerosol transmission infect sheep, goats, people, camels, cattle, buffaloes, and so on. In this paper, a conserved region of the S RNA segment of Rift Valley fever virus (RVFV) ZH501 strain w...
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2020.590732/full |
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| author | Qiuxue Han Qiuxue Han Shengnan Zhang Dongping Liu Feihu Yan Feihu Yan Hualei Wang Pei Huang Pei Huang Jinhao Bi Jinhao Bi Hongli Jin Hongli Jin Na Feng Zengguo Cao Zengguo Cao Yuwei Gao Yuwei Gao Hang Chi Hang Chi Songtao Yang Songtao Yang Yongkun Zhao Yongkun Zhao Xianzhu Xia Xianzhu Xia Xianzhu Xia |
| author_facet | Qiuxue Han Qiuxue Han Shengnan Zhang Dongping Liu Feihu Yan Feihu Yan Hualei Wang Pei Huang Pei Huang Jinhao Bi Jinhao Bi Hongli Jin Hongli Jin Na Feng Zengguo Cao Zengguo Cao Yuwei Gao Yuwei Gao Hang Chi Hang Chi Songtao Yang Songtao Yang Yongkun Zhao Yongkun Zhao Xianzhu Xia Xianzhu Xia Xianzhu Xia |
| author_sort | Qiuxue Han |
| collection | DOAJ |
| description | Rift Valley fever (RVF) is a severe infectious disease, which can through mosquito bites, direct contact and aerosol transmission infect sheep, goats, people, camels, cattle, buffaloes, and so on. In this paper, a conserved region of the S RNA segment of Rift Valley fever virus (RVFV) ZH501 strain was used as target sequence. The RVFV RT-LAMP-VF assay was successfully established combined reverse transcription-loop-mediated isothermal amplification with a vertical flow visualization strip. The detection limit is up to 1.94 × 100 copies/μl of synthesized RVFV-RNA. RNA extracted from cell culture of an inactivated RVFV-BJ01 strain was also used as templates, and the detection limit is 1.83 × 103 copies/μl. In addition, there was no cross-reactivity with other viruses that can cause similar fever symptoms. The RVFV-LAMP-VF assay exhibited very high levels of diagnostic sensitivity, which had 100-fold more sensitive than RVFV real-time RT-PCR assay. Accordingly, the RVFV RT-LAMP-VF assay developed in this study is suitable for the rapid and sensitive diagnosis of RVFV without specialized equipment and can rapidly complete detection within 60 min, and the results are visible by vertical flow visualization strip within 5 min. |
| first_indexed | 2024-12-22T20:27:13Z |
| format | Article |
| id | doaj.art-cb5a12cea0e246fbad89900538b4ebb3 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-22T20:27:13Z |
| publishDate | 2020-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-cb5a12cea0e246fbad89900538b4ebb32022-12-21T18:13:42ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-11-011110.3389/fmicb.2020.590732590732Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever VirusQiuxue Han0Qiuxue Han1Shengnan Zhang2Dongping Liu3Feihu Yan4Feihu Yan5Hualei Wang6Pei Huang7Pei Huang8Jinhao Bi9Jinhao Bi10Hongli Jin11Hongli Jin12Na Feng13Zengguo Cao14Zengguo Cao15Yuwei Gao16Yuwei Gao17Hang Chi18Hang Chi19Songtao Yang20Songtao Yang21Yongkun Zhao22Yongkun Zhao23Xianzhu Xia24Xianzhu Xia25Xianzhu Xia26Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS) and Comparative Medicine Center, Peking Union Medical College (PUMC), Beijing, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaThe Nanjing Unicorn Academy of Innovation, Institute Pasteur of Shanghai, Chinese Academy of Sciences, Nanjing, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, ChinaCollege of Veterinary Medicine, Jilin University, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaAnimal Science and Technology College, Jilin Agricultural University, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaAnimal Science and Technology College, Jilin Agricultural University, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaCollege of Veterinary Medicine, Jilin University, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaCollege of Veterinary Medicine, Jilin University, Changchun, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, ChinaInstitute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS) and Comparative Medicine Center, Peking Union Medical College (PUMC), Beijing, ChinaKey Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, ChinaRift Valley fever (RVF) is a severe infectious disease, which can through mosquito bites, direct contact and aerosol transmission infect sheep, goats, people, camels, cattle, buffaloes, and so on. In this paper, a conserved region of the S RNA segment of Rift Valley fever virus (RVFV) ZH501 strain was used as target sequence. The RVFV RT-LAMP-VF assay was successfully established combined reverse transcription-loop-mediated isothermal amplification with a vertical flow visualization strip. The detection limit is up to 1.94 × 100 copies/μl of synthesized RVFV-RNA. RNA extracted from cell culture of an inactivated RVFV-BJ01 strain was also used as templates, and the detection limit is 1.83 × 103 copies/μl. In addition, there was no cross-reactivity with other viruses that can cause similar fever symptoms. The RVFV-LAMP-VF assay exhibited very high levels of diagnostic sensitivity, which had 100-fold more sensitive than RVFV real-time RT-PCR assay. Accordingly, the RVFV RT-LAMP-VF assay developed in this study is suitable for the rapid and sensitive diagnosis of RVFV without specialized equipment and can rapidly complete detection within 60 min, and the results are visible by vertical flow visualization strip within 5 min.https://www.frontiersin.org/articles/10.3389/fmicb.2020.590732/fullRift Valley fever virusreverse transcription-loop-mediated isothermal amplificationnucleic acid visualizationvisual detectioninactivated RVFV-BJ01 strain |
| spellingShingle | Qiuxue Han Qiuxue Han Shengnan Zhang Dongping Liu Feihu Yan Feihu Yan Hualei Wang Pei Huang Pei Huang Jinhao Bi Jinhao Bi Hongli Jin Hongli Jin Na Feng Zengguo Cao Zengguo Cao Yuwei Gao Yuwei Gao Hang Chi Hang Chi Songtao Yang Songtao Yang Yongkun Zhao Yongkun Zhao Xianzhu Xia Xianzhu Xia Xianzhu Xia Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus Frontiers in Microbiology Rift Valley fever virus reverse transcription-loop-mediated isothermal amplification nucleic acid visualization visual detection inactivated RVFV-BJ01 strain |
| title | Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus |
| title_full | Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus |
| title_fullStr | Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus |
| title_full_unstemmed | Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus |
| title_short | Development of a Visible Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for the Detection of Rift Valley Fever Virus |
| title_sort | development of a visible reverse transcription loop mediated isothermal amplification assay for the detection of rift valley fever virus |
| topic | Rift Valley fever virus reverse transcription-loop-mediated isothermal amplification nucleic acid visualization visual detection inactivated RVFV-BJ01 strain |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2020.590732/full |
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