A Rapid and Specific Assay for the Detection of MERS-CoV
Identifieur interne : 000C81 ( Pmc/Curation ); précédent : 000C80; suivant : 000C82A Rapid and Specific Assay for the Detection of MERS-CoV
Auteurs : Pei Huang [République populaire de Chine] ; Hualei Wang [République populaire de Chine] ; Zengguo Cao [République populaire de Chine] ; Hongli Jin [République populaire de Chine] ; Hang Chi [République populaire de Chine] ; Jincun Zhao [République populaire de Chine] ; Beibei Yu [République populaire de Chine] ; Feihu Yan [République populaire de Chine] ; Xingxing Hu [République populaire de Chine] ; Fangfang Wu [République populaire de Chine] ; Cuicui Jiao [République populaire de Chine] ; Pengfei Hou [République populaire de Chine] ; Shengnan Xu [République populaire de Chine] ; Yongkun Zhao [République populaire de Chine] ; Na Feng [République populaire de Chine] ; Jianzhong Wang [République populaire de Chine] ; Weiyang Sun [République populaire de Chine] ; Tiecheng Wang [République populaire de Chine] ; Yuwei Gao [République populaire de Chine] ; Songtao Yang [République populaire de Chine] ; Xianzhu Xia [République populaire de Chine]Source :
- Frontiers in Microbiology [ 1664-302X ] ; 2018.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel human coronavirus that can cause human respiratory disease. The development of a detection method for this virus that can lead to rapid and accurate diagnosis would be significant. In this study, we established a nucleic acid visualization technique that combines the reverse transcription loop-mediated isothermal amplification technique and a vertical flow visualization strip (RT-LAMP-VF) to detect the N gene of MERS-CoV. The RT-LAMP-VF assay was performed in a constant temperature water bath for 30 min, and the result was visible by the naked eye within 5 min. The RT-LAMP-VF assay was capable of detecting 2 × 101 copies/μl of synthesized RNA transcript and 1 × 101 copies/μl of MERS-CoV RNA. The method exhibits no cross-reactivities with multiple CoVs including SARS-related (SARSr)-CoV, HKU4, HKU1, OC43 and 229E, and thus exhibits high specificity. Compared to the real-time RT-PCR (rRT-PCR) method recommended by the World Health Organization (WHO), the RT-LAMP-VF assay is easy to handle, does not require expensive equipment and can rapidly complete detection within 35 min.
Url:
DOI: 10.3389/fmicb.2018.01101
PubMed: 29896174
PubMed Central: 5987675
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<author><name sortKey="Sun, Weiyang" sort="Sun, Weiyang" uniqKey="Sun W" first="Weiyang" last="Sun">Weiyang Sun</name>
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<author><name sortKey="Wang, Tiecheng" sort="Wang, Tiecheng" uniqKey="Wang T" first="Tiecheng" last="Wang">Tiecheng Wang</name>
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<author><name sortKey="Gao, Yuwei" sort="Gao, Yuwei" uniqKey="Gao Y" first="Yuwei" last="Gao">Yuwei Gao</name>
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<author><name sortKey="Yang, Songtao" sort="Yang, Songtao" uniqKey="Yang S" first="Songtao" last="Yang">Songtao Yang</name>
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<author><name sortKey="Xia, Xianzhu" sort="Xia, Xianzhu" uniqKey="Xia X" first="Xianzhu" last="Xia">Xianzhu Xia</name>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A Rapid and Specific Assay for the Detection of MERS-CoV</title>
<author><name sortKey="Huang, Pei" sort="Huang, Pei" uniqKey="Huang P" first="Pei" last="Huang">Pei Huang</name>
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</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<author><name sortKey="Wang, Hualei" sort="Wang, Hualei" uniqKey="Wang H" first="Hualei" last="Wang">Hualei Wang</name>
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<country xml:lang="fr">République populaire de Chine</country>
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<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
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</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
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</author>
<author><name sortKey="Cao, Zengguo" sort="Cao, Zengguo" uniqKey="Cao Z" first="Zengguo" last="Cao">Zengguo Cao</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>College of Veterinary Medicine, Jilin University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Jin, Hongli" sort="Jin, Hongli" uniqKey="Jin H" first="Hongli" last="Jin">Hongli Jin</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>College of Veterinary Medicine, Jilin University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Chi, Hang" sort="Chi, Hang" uniqKey="Chi H" first="Hang" last="Chi">Hang Chi</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Zhao, Jincun" sort="Zhao, Jincun" uniqKey="Zhao J" first="Jincun" last="Zhao">Jincun Zhao</name>
<affiliation wicri:level="1"><nlm:aff id="aff5"><institution>State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Heath, The First Affiliated Hospital of Guangzhou Medical University</institution>
,<addr-line>Guangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff6"><institution>Guangzhou Eighth People’s Hospital of Guangzhou Medical University</institution>
,<addr-line>Guangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Yu, Beibei" sort="Yu, Beibei" uniqKey="Yu B" first="Beibei" last="Yu">Beibei Yu</name>
<affiliation wicri:level="1"><nlm:aff id="aff7"><institution>Department of Clinical Laboratory, College of Medicine, Sir Run Run Shaw Hospital, Zhejiang University</institution>
,<addr-line>Hangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Yan, Feihu" sort="Yan, Feihu" uniqKey="Yan F" first="Feihu" last="Yan">Feihu Yan</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Hu, Xingxing" sort="Hu, Xingxing" uniqKey="Hu X" first="Xingxing" last="Hu">Xingxing Hu</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Animal Science and Technology College, Jilin Agricultural University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Wu, Fangfang" sort="Wu, Fangfang" uniqKey="Wu F" first="Fangfang" last="Wu">Fangfang Wu</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Jiao, Cuicui" sort="Jiao, Cuicui" uniqKey="Jiao C" first="Cuicui" last="Jiao">Cuicui Jiao</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Hou, Pengfei" sort="Hou, Pengfei" uniqKey="Hou P" first="Pengfei" last="Hou">Pengfei Hou</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff3"><institution>College of Veterinary Medicine, Jilin University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Xu, Shengnan" sort="Xu, Shengnan" uniqKey="Xu S" first="Shengnan" last="Xu">Shengnan Xu</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Animal Science and Technology College, Jilin Agricultural University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Zhao, Yongkun" sort="Zhao, Yongkun" uniqKey="Zhao Y" first="Yongkun" last="Zhao">Yongkun Zhao</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Feng, Na" sort="Feng, Na" uniqKey="Feng N" first="Na" last="Feng">Na Feng</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Wang, Jianzhong" sort="Wang, Jianzhong" uniqKey="Wang J" first="Jianzhong" last="Wang">Jianzhong Wang</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Animal Science and Technology College, Jilin Agricultural University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Sun, Weiyang" sort="Sun, Weiyang" uniqKey="Sun W" first="Weiyang" last="Sun">Weiyang Sun</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Wang, Tiecheng" sort="Wang, Tiecheng" uniqKey="Wang T" first="Tiecheng" last="Wang">Tiecheng Wang</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Gao, Yuwei" sort="Gao, Yuwei" uniqKey="Gao Y" first="Yuwei" last="Gao">Yuwei Gao</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Yang, Songtao" sort="Yang, Songtao" uniqKey="Yang S" first="Songtao" last="Yang">Songtao Yang</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Xia, Xianzhu" sort="Xia, Xianzhu" uniqKey="Xia X" first="Xianzhu" last="Xia">Xianzhu Xia</name>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series><title level="j">Frontiers in Microbiology</title>
<idno type="eISSN">1664-302X</idno>
<imprint><date when="2018">2018</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel human coronavirus that can cause human respiratory disease. The development of a detection method for this virus that can lead to rapid and accurate diagnosis would be significant. In this study, we established a nucleic acid visualization technique that combines the reverse transcription loop-mediated isothermal amplification technique and a vertical flow visualization strip (RT-LAMP-VF) to detect the N gene of MERS-CoV. The RT-LAMP-VF assay was performed in a constant temperature water bath for 30 min, and the result was visible by the naked eye within 5 min. The RT-LAMP-VF assay was capable of detecting 2 × 10<sup>1</sup>
copies/μl of synthesized RNA transcript and 1 × 10<sup>1</sup>
copies/μl of MERS-CoV RNA. The method exhibits no cross-reactivities with multiple CoVs including SARS-related (SARSr)-CoV, HKU4, HKU1, OC43 and 229E, and thus exhibits high specificity. Compared to the real-time RT-PCR (rRT-PCR) method recommended by the World Health Organization (WHO), the RT-LAMP-VF assay is easy to handle, does not require expensive equipment and can rapidly complete detection within 35 min.</p>
</div>
</front>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Front Microbiol</journal-id>
<journal-id journal-id-type="iso-abbrev">Front Microbiol</journal-id>
<journal-id journal-id-type="publisher-id">Front. Microbiol.</journal-id>
<journal-title-group><journal-title>Frontiers in Microbiology</journal-title>
</journal-title-group>
<issn pub-type="epub">1664-302X</issn>
<publisher><publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">29896174</article-id>
<article-id pub-id-type="pmc">5987675</article-id>
<article-id pub-id-type="doi">10.3389/fmicb.2018.01101</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Microbiology</subject>
<subj-group><subject>Original Research</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group><article-title>A Rapid and Specific Assay for the Detection of MERS-CoV</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Huang</surname>
<given-names>Pei</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Hualei</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
<xref ref-type="corresp" rid="c001"><sup>*</sup>
</xref>
<uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/200626/overview"></uri>
</contrib>
<contrib contrib-type="author"><name><surname>Cao</surname>
<given-names>Zengguo</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Jin</surname>
<given-names>Hongli</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chi</surname>
<given-names>Hang</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhao</surname>
<given-names>Jincun</given-names>
</name>
<xref ref-type="aff" rid="aff5"><sup>5</sup>
</xref>
<xref ref-type="aff" rid="aff6"><sup>6</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yu</surname>
<given-names>Beibei</given-names>
</name>
<xref ref-type="aff" rid="aff7"><sup>7</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yan</surname>
<given-names>Feihu</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Hu</surname>
<given-names>Xingxing</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wu</surname>
<given-names>Fangfang</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Jiao</surname>
<given-names>Cuicui</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Hou</surname>
<given-names>Pengfei</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff3"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Xu</surname>
<given-names>Shengnan</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhao</surname>
<given-names>Yongkun</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Feng</surname>
<given-names>Na</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Jianzhong</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Sun</surname>
<given-names>Weiyang</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Wang</surname>
<given-names>Tiecheng</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Gao</surname>
<given-names>Yuwei</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
<uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/234182/overview"></uri>
</contrib>
<contrib contrib-type="author"><name><surname>Yang</surname>
<given-names>Songtao</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
<uri xlink:type="simple" xlink:href="http://loop.frontiersin.org/people/424974/overview"></uri>
</contrib>
<contrib contrib-type="author"><name><surname>Xia</surname>
<given-names>Xianzhu</given-names>
</name>
<xref ref-type="aff" rid="aff2"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="aff4"><sup>4</sup>
</xref>
</contrib>
</contrib-group>
<aff id="aff1"><sup>1</sup>
<institution>Animal Science and Technology College, Jilin Agricultural University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</aff>
<aff id="aff2"><sup>2</sup>
<institution>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</aff>
<aff id="aff3"><sup>3</sup>
<institution>College of Veterinary Medicine, Jilin University</institution>
,<addr-line>Changchun</addr-line>
,<country>China</country>
</aff>
<aff id="aff4"><sup>4</sup>
<institution>Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses</institution>
,<addr-line>Yangzhou</addr-line>
,<country>China</country>
</aff>
<aff id="aff5"><sup>5</sup>
<institution>State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Heath, The First Affiliated Hospital of Guangzhou Medical University</institution>
,<addr-line>Guangzhou</addr-line>
,<country>China</country>
</aff>
<aff id="aff6"><sup>6</sup>
<institution>Guangzhou Eighth People’s Hospital of Guangzhou Medical University</institution>
,<addr-line>Guangzhou</addr-line>
,<country>China</country>
</aff>
<aff id="aff7"><sup>7</sup>
<institution>Department of Clinical Laboratory, College of Medicine, Sir Run Run Shaw Hospital, Zhejiang University</institution>
,<addr-line>Hangzhou</addr-line>
,<country>China</country>
</aff>
<author-notes><fn fn-type="edited-by"><p>Edited by: Dirk Dittmer, University of North Carolina at Chapel Hill, United States</p>
</fn>
<fn fn-type="edited-by"><p>Reviewed by: Timothy Sheahan, University of North Carolina at Chapel Hill, United States; Yize Li, University of Pennsylvania, United States</p>
</fn>
<corresp id="c001">*Correspondence: Hualei Wang, <email>whl831125@163.com</email>
; <email>wangh25@hotmail.com</email>
</corresp>
<fn fn-type="other" id="fn002"><p>This article was submitted to Virology, a section of the journal Frontiers in Microbiology</p>
</fn>
</author-notes>
<pub-date pub-type="epub"><day>29</day>
<month>5</month>
<year>2018</year>
</pub-date>
<pub-date pub-type="collection"><year>2018</year>
</pub-date>
<volume>9</volume>
<elocation-id>1101</elocation-id>
<history><date date-type="received"><day>30</day>
<month>12</month>
<year>2017</year>
</date>
<date date-type="accepted"><day>08</day>
<month>5</month>
<year>2018</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2018 Huang, Wang, Cao, Jin, Chi, Zhao, Yu, Yan, Hu, Wu, Jiao, Hou, Xu, Zhao, Feng, Wang, Sun, Wang, Gao, Yang and Xia.</copyright-statement>
<copyright-year>2018</copyright-year>
<copyright-holder>Huang, Wang, Cao, Jin, Chi, Zhao, Yu, Yan, Hu, Wu, Jiao, Hou, Xu, Zhao, Feng, Wang, Sun, Wang, Gao, Yang and Xia</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</license-p>
</license>
</permissions>
<abstract><p>Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel human coronavirus that can cause human respiratory disease. The development of a detection method for this virus that can lead to rapid and accurate diagnosis would be significant. In this study, we established a nucleic acid visualization technique that combines the reverse transcription loop-mediated isothermal amplification technique and a vertical flow visualization strip (RT-LAMP-VF) to detect the N gene of MERS-CoV. The RT-LAMP-VF assay was performed in a constant temperature water bath for 30 min, and the result was visible by the naked eye within 5 min. The RT-LAMP-VF assay was capable of detecting 2 × 10<sup>1</sup>
copies/μl of synthesized RNA transcript and 1 × 10<sup>1</sup>
copies/μl of MERS-CoV RNA. The method exhibits no cross-reactivities with multiple CoVs including SARS-related (SARSr)-CoV, HKU4, HKU1, OC43 and 229E, and thus exhibits high specificity. Compared to the real-time RT-PCR (rRT-PCR) method recommended by the World Health Organization (WHO), the RT-LAMP-VF assay is easy to handle, does not require expensive equipment and can rapidly complete detection within 35 min.</p>
</abstract>
<kwd-group><kwd>Middle East respiratory syndrome coronavirus</kwd>
<kwd>reverse transcription loop-mediated isothermal amplification</kwd>
<kwd>nucleic acid visualization</kwd>
<kwd>visual detection</kwd>
<kwd>RT-LAMP-VF</kwd>
</kwd-group>
<counts><fig-count count="6"></fig-count>
<table-count count="5"></table-count>
<equation-count count="0"></equation-count>
<ref-count count="26"></ref-count>
<page-count count="9"></page-count>
<word-count count="0"></word-count>
</counts>
</article-meta>
</front>
</pmc>
</record>
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