A Rapid and Specific Assay for the Detection of MERS-CoV.
Identifieur interne : 000D22 ( Main/Exploration ); précédent : 000D21; suivant : 000D23A 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.
DOI: 10.3389/fmicb.2018.01101
PubMed: 29896174
Affiliations:
- République populaire de Chine
- Dongbei, Guangdong, Jilin, Zhejiang
- Changchun, Hangzhou, Jiangmen
- Université de Zhejiang
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Xianzhu" sort="Xia, Xianzhu" uniqKey="Xia X" first="Xianzhu" last="Xia">Xianzhu Xia</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2018">2018</date><idno type="RBID">pubmed:29896174</idno><idno type="pmid">29896174</idno><idno type="doi">10.3389/fmicb.2018.01101</idno><idno 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uniqKey="Huang P" first="Pei" last="Huang">Pei Huang</name><affiliation wicri:level="3"><nlm:affiliation>Animal Science and Technology College, Jilin Agricultural University, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Animal Science and Technology College, Jilin Agricultural University, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Wang, Hualei" sort="Wang, Hualei" uniqKey="Wang H" first="Hualei" last="Wang">Hualei Wang</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Cao, Zengguo" sort="Cao, Zengguo" uniqKey="Cao Z" first="Zengguo" last="Cao">Zengguo Cao</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Jin, Hongli" sort="Jin, Hongli" uniqKey="Jin H" first="Hongli" last="Jin">Hongli Jin</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Chi, Hang" sort="Chi, Hang" uniqKey="Chi H" first="Hang" last="Chi">Hang Chi</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Zhao, Jincun" sort="Zhao, Jincun" uniqKey="Zhao J" first="Jincun" last="Zhao">Jincun Zhao</name><affiliation wicri:level="3"><nlm:affiliation>State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Heath, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Heath, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Yu, Beibei" sort="Yu, Beibei" uniqKey="Yu B" first="Beibei" last="Yu">Beibei Yu</name><affiliation wicri:level="4"><nlm:affiliation>Department of Clinical Laboratory, College of Medicine, Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Clinical Laboratory, College of Medicine, Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou</wicri:regionArea><placeName><settlement type="city">Hangzhou</settlement><region type="province">Zhejiang</region></placeName><orgName type="university">Université de Zhejiang</orgName></affiliation></author><author><name sortKey="Yan, Feihu" sort="Yan, Feihu" uniqKey="Yan F" first="Feihu" last="Yan">Feihu Yan</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Hu, Xingxing" sort="Hu, Xingxing" uniqKey="Hu X" first="Xingxing" last="Hu">Xingxing Hu</name><affiliation wicri:level="3"><nlm:affiliation>Animal Science and Technology College, Jilin Agricultural University, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Animal Science and Technology College, Jilin Agricultural University, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Wu, Fangfang" sort="Wu, Fangfang" uniqKey="Wu F" first="Fangfang" last="Wu">Fangfang Wu</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Jiao, Cuicui" sort="Jiao, Cuicui" uniqKey="Jiao C" first="Cuicui" last="Jiao">Cuicui Jiao</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Hou, Pengfei" sort="Hou, Pengfei" uniqKey="Hou P" first="Pengfei" last="Hou">Pengfei Hou</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Xu, Shengnan" sort="Xu, Shengnan" uniqKey="Xu S" first="Shengnan" last="Xu">Shengnan Xu</name><affiliation wicri:level="3"><nlm:affiliation>Animal Science and Technology College, Jilin Agricultural University, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Animal Science and Technology College, Jilin Agricultural University, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Zhao, Yongkun" sort="Zhao, Yongkun" uniqKey="Zhao Y" first="Yongkun" last="Zhao">Yongkun Zhao</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Feng, Na" sort="Feng, Na" uniqKey="Feng N" first="Na" last="Feng">Na Feng</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Wang, Jianzhong" sort="Wang, Jianzhong" uniqKey="Wang J" first="Jianzhong" last="Wang">Jianzhong Wang</name><affiliation wicri:level="3"><nlm:affiliation>Animal Science and Technology College, Jilin Agricultural University, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Animal Science and Technology College, Jilin Agricultural University, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Sun, Weiyang" sort="Sun, Weiyang" uniqKey="Sun W" first="Weiyang" last="Sun">Weiyang Sun</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Wang, Tiecheng" sort="Wang, Tiecheng" uniqKey="Wang T" first="Tiecheng" last="Wang">Tiecheng Wang</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Gao, Yuwei" sort="Gao, Yuwei" uniqKey="Gao Y" first="Yuwei" last="Gao">Yuwei Gao</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Yang, Songtao" sort="Yang, Songtao" uniqKey="Yang S" first="Songtao" last="Yang">Songtao Yang</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author><author><name sortKey="Xia, Xianzhu" sort="Xia, Xianzhu" uniqKey="Xia X" first="Xianzhu" last="Xia">Xianzhu Xia</name><affiliation wicri:level="3"><nlm:affiliation>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun</wicri:regionArea><placeName><settlement type="city">Changchun</settlement><region type="province">Jilin</region><region type="groupement">Dongbei</region></placeName></affiliation></author></analytic><series><title level="j">Frontiers in microbiology</title><idno type="ISSN">1664-302X</idno><imprint><date when="2018" type="published">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Dongbei</li><li>Guangdong</li><li>Jilin</li><li>Zhejiang</li></region><settlement><li>Changchun</li><li>Hangzhou</li><li>Jiangmen</li></settlement><orgName><li>Université de Zhejiang</li></orgName></list><tree><country name="République populaire de Chine"><region name="Jilin"><name sortKey="Huang, Pei" sort="Huang, Pei" uniqKey="Huang P" first="Pei" last="Huang">Pei Huang</name></region><name sortKey="Cao, Zengguo" sort="Cao, Zengguo" uniqKey="Cao Z" first="Zengguo" last="Cao">Zengguo Cao</name><name sortKey="Chi, Hang" sort="Chi, Hang" uniqKey="Chi H" first="Hang" last="Chi">Hang Chi</name><name sortKey="Feng, Na" sort="Feng, Na" uniqKey="Feng N" first="Na" last="Feng">Na Feng</name><name sortKey="Gao, Yuwei" sort="Gao, Yuwei" uniqKey="Gao Y" first="Yuwei" last="Gao">Yuwei Gao</name><name sortKey="Hou, Pengfei" sort="Hou, Pengfei" uniqKey="Hou P" first="Pengfei" last="Hou">Pengfei Hou</name><name sortKey="Hu, Xingxing" sort="Hu, Xingxing" uniqKey="Hu X" first="Xingxing" last="Hu">Xingxing Hu</name><name sortKey="Jiao, Cuicui" sort="Jiao, Cuicui" uniqKey="Jiao C" first="Cuicui" last="Jiao">Cuicui Jiao</name><name sortKey="Jin, Hongli" sort="Jin, Hongli" uniqKey="Jin H" first="Hongli" last="Jin">Hongli Jin</name><name sortKey="Sun, Weiyang" sort="Sun, Weiyang" uniqKey="Sun W" first="Weiyang" last="Sun">Weiyang Sun</name><name sortKey="Wang, Hualei" sort="Wang, Hualei" uniqKey="Wang H" first="Hualei" last="Wang">Hualei Wang</name><name sortKey="Wang, Jianzhong" sort="Wang, Jianzhong" uniqKey="Wang J" first="Jianzhong" last="Wang">Jianzhong Wang</name><name sortKey="Wang, Tiecheng" sort="Wang, Tiecheng" uniqKey="Wang T" first="Tiecheng" last="Wang">Tiecheng Wang</name><name sortKey="Wu, Fangfang" sort="Wu, Fangfang" uniqKey="Wu F" first="Fangfang" last="Wu">Fangfang Wu</name><name sortKey="Xia, Xianzhu" sort="Xia, Xianzhu" uniqKey="Xia X" first="Xianzhu" last="Xia">Xianzhu Xia</name><name sortKey="Xu, Shengnan" sort="Xu, Shengnan" uniqKey="Xu S" first="Shengnan" last="Xu">Shengnan Xu</name><name sortKey="Yan, Feihu" sort="Yan, Feihu" uniqKey="Yan F" first="Feihu" last="Yan">Feihu Yan</name><name sortKey="Yang, Songtao" sort="Yang, Songtao" uniqKey="Yang S" first="Songtao" last="Yang">Songtao Yang</name><name sortKey="Yu, Beibei" sort="Yu, Beibei" uniqKey="Yu B" first="Beibei" last="Yu">Beibei Yu</name><name sortKey="Zhao, Jincun" sort="Zhao, Jincun" uniqKey="Zhao J" first="Jincun" last="Zhao">Jincun Zhao</name><name sortKey="Zhao, Yongkun" sort="Zhao, Yongkun" uniqKey="Zhao Y" first="Yongkun" last="Zhao">Yongkun Zhao</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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