A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses
Identifieur interne : 001C77 ( Ncbi/Merge ); précédent : 001C76; suivant : 001C78A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses
Auteurs : Yong Yan [République populaire de Chine] ; Jian-Yong Luo [République populaire de Chine] ; Yin Chen [République populaire de Chine] ; Heng-Hui Wang [République populaire de Chine] ; Guo-Ying Zhu [République populaire de Chine] ; Pei-Yan He [République populaire de Chine] ; Jin-Lei Guo [République populaire de Chine] ; Yong-Liang Lei [République populaire de Chine] ; Zhong-Wen Chen [République populaire de Chine]Source :
- Oncotarget [ 1949-2553 ] ; 2017.
Abstract
We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.10% for FluA, 94.59% for FluB, 98.68% for PIV-3, 94.87% for RSV and 95.92% for MPV (No Data for MERS-CoV due to the lack of positive specimens). Data of analytical sensitivities showed that the detection limits of the rMLA assay were 5–25 viral RNA copies per μl for FluA, FluB, PIV-3 and MERS-CoV, approximate to the real-time RT-PCR assay; while the values were 8 and 22copies/μl for MPV and RSV, lower than the real-time RT-PCR(78 and 114 copies/μl respectively). The results indicated that the rMLA is a sensitive, specific detection tool and comparable to real-time RT-PCR, especially suitable for high-throughput detection of respiratory specimens.
Url:
DOI: 10.18632/oncotarget.18533
PubMed: 29228581
PubMed Central: 5722533
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<front><div type="abstract" xml:lang="en"><p>We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.10% for FluA, 94.59% for FluB, 98.68% for PIV-3, 94.87% for RSV and 95.92% for MPV (No Data for MERS-CoV due to the lack of positive specimens). Data of analytical sensitivities showed that the detection limits of the rMLA assay were 5–25 viral RNA copies per μl for FluA, FluB, PIV-3 and MERS-CoV, approximate to the real-time RT-PCR assay; while the values were 8 and 22copies/μl for MPV and RSV, lower than the real-time RT-PCR(78 and 114 copies/μl respectively). The results indicated that the rMLA is a sensitive, specific detection tool and comparable to real-time RT-PCR, especially suitable for high-throughput detection of respiratory specimens.</p>
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<affiliation wicri:level="1"><nlm:aff id="A3"> Lishui Municipal Center for Disease Control and Prevention, Lishui 323000, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Lishui Municipal Center for Disease Control and Prevention, Lishui 323000</wicri:regionArea>
<wicri:noRegion>Lishui 323000</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chen, Zhong Wen" sort="Chen, Zhong Wen" uniqKey="Chen Z" first="Zhong-Wen" last="Chen">Zhong-Wen Chen</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
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<idno type="pmid">29228581</idno>
<idno type="pmc">5722533</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5722533</idno>
<idno type="RBID">PMC:5722533</idno>
<idno type="doi">10.18632/oncotarget.18533</idno>
<date when="2017">2017</date>
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<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000946</idno>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses</title>
<author><name sortKey="Yan, Yong" sort="Yan, Yong" uniqKey="Yan Y" first="Yong" last="Yan">Yong Yan</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Luo, Jian Yong" sort="Luo, Jian Yong" uniqKey="Luo J" first="Jian-Yong" last="Luo">Jian-Yong Luo</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chen, Yin" sort="Chen, Yin" uniqKey="Chen Y" first="Yin" last="Chen">Yin Chen</name>
<affiliation wicri:level="1"><nlm:aff id="A2"> Institute of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Institute of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051</wicri:regionArea>
<placeName><settlement type="city">Hangzhou</settlement>
<region type="province">Zhejiang</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Wang, Heng Hui" sort="Wang, Heng Hui" uniqKey="Wang H" first="Heng-Hui" last="Wang">Heng-Hui Wang</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Zhu, Guo Ying" sort="Zhu, Guo Ying" uniqKey="Zhu G" first="Guo-Ying" last="Zhu">Guo-Ying Zhu</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="He, Pei Yan" sort="He, Pei Yan" uniqKey="He P" first="Pei-Yan" last="He">Pei-Yan He</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Guo, Jin Lei" sort="Guo, Jin Lei" uniqKey="Guo J" first="Jin-Lei" last="Guo">Jin-Lei Guo</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Lei, Yong Liang" sort="Lei, Yong Liang" uniqKey="Lei Y" first="Yong-Liang" last="Lei">Yong-Liang Lei</name>
<affiliation wicri:level="1"><nlm:aff id="A3"> Lishui Municipal Center for Disease Control and Prevention, Lishui 323000, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Lishui Municipal Center for Disease Control and Prevention, Lishui 323000</wicri:regionArea>
<wicri:noRegion>Lishui 323000</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chen, Zhong Wen" sort="Chen, Zhong Wen" uniqKey="Chen Z" first="Zhong-Wen" last="Chen">Zhong-Wen Chen</name>
<affiliation wicri:level="1"><nlm:aff id="A1"> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050, China</nlm:aff>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea> Jiaxing Key Laboratory of Pathogenic Microbiology, Jiaxing Municipal Center for Disease Control and Prevention, Jiaxing 314050</wicri:regionArea>
<wicri:noRegion>Jiaxing 314050</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">Oncotarget</title>
<idno type="eISSN">1949-2553</idno>
<imprint><date when="2017">2017</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.10% for FluA, 94.59% for FluB, 98.68% for PIV-3, 94.87% for RSV and 95.92% for MPV (No Data for MERS-CoV due to the lack of positive specimens). Data of analytical sensitivities showed that the detection limits of the rMLA assay were 5–25 viral RNA copies per μl for FluA, FluB, PIV-3 and MERS-CoV, approximate to the real-time RT-PCR assay; while the values were 8 and 22copies/μl for MPV and RSV, lower than the real-time RT-PCR(78 and 114 copies/μl respectively). The results indicated that the rMLA is a sensitive, specific detection tool and comparable to real-time RT-PCR, especially suitable for high-throughput detection of respiratory specimens.</p>
</div>
</front>
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<author><name sortKey="Wang, Heng Hui" sort="Wang, Heng Hui" uniqKey="Wang H" first="Heng-Hui" last="Wang">Heng-Hui Wang</name>
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<country xml:lang="fr">République populaire de Chine</country>
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</affiliation>
</author>
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<country xml:lang="fr">République populaire de Chine</country>
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<author><name sortKey="He, Pei Yan" sort="He, Pei Yan" uniqKey="He P" first="Pei-Yan" last="He">Pei-Yan He</name>
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<author><name sortKey="Guo, Jin Lei" sort="Guo, Jin Lei" uniqKey="Guo J" first="Jin-Lei" last="Guo">Jin-Lei Guo</name>
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<author><name sortKey="Lei, Yong Liang" sort="Lei, Yong Liang" uniqKey="Lei Y" first="Yong-Liang" last="Lei">Yong-Liang Lei</name>
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<author><name sortKey="Chen, Zhong Wen" sort="Chen, Zhong Wen" uniqKey="Chen Z" first="Zhong-Wen" last="Chen">Zhong-Wen Chen</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>
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<front><div type="abstract" xml:lang="en">We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.10% for FluA, 94.59% for FluB, 98.68% for PIV-3, 94.87% for RSV and 95.92% for MPV (No Data for MERS-CoV due to the lack of positive specimens). Data of analytical sensitivities showed that the detection limits of the rMLA assay were 5-25 viral RNA copies per μl for FluA, FluB, PIV-3 and MERS-CoV, approximate to the real-time RT-PCR assay; while the values were 8 and 22copies/μl for MPV and RSV, lower than the real-time RT-PCR(78 and 114 copies/μl respectively). The results indicated that the rMLA is a sensitive, specific detection tool and comparable to real-time RT-PCR, especially suitable for high-throughput detection of respiratory specimens.</div>
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