A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals
Identifieur interne : 000726 ( Main/Exploration ); précédent : 000725; suivant : 000727A novel luciferase immunosorbent assay performs better than a commercial enzyme-linked immunosorbent assay to detect MERS-CoV specific IgG in humans and animals
Auteurs : Wenling Wang [République populaire de Chine] ; Tianyu Wang [République populaire de Chine] ; Yao Deng [République populaire de Chine] ; Peihua Niu [République populaire de Chine] ; Ruhan A [République populaire de Chine] ; Jincun Zhao [République populaire de Chine] ; Malik Peiris ; Shixing Tang [République populaire de Chine] ; Wenjie Tan [République populaire de Chine]Source :
- Biosafety and Health [ 2590-0536 ] ; 2019.
Abstract
The Middle East respiratory syndrome (MERS) is a lethal zoonosis caused by MERS coronavirus (MERS-CoV) and poses a significant threat to public health worldwide. Therefore, a rapid, sensitive, and specific serologic test for detecting anti-MERS-CoV antibodies in both humans and animals is urgently needed for the successful management of this illness. Here, we evaluated various novel luciferase immunosorbent assays (LISA) based on nucleocapsid protein (NP) as well as fragments derived from spike protein (S) including subunit 1 (S1), N terminal domain (NTD), receptor-binding domain (RBD) and subunit 2 (S2) of S for the detection of MERS-CoV-specific IgG. Fusion proteins, including nanoluciferase (NLuc) and various fragments derived from the NP or S protein of MERS-CoV, were expressed in human embryonic kidney 293 T cells. LISAs that detected anti-MERS-CoV IgG were further developed using cell lysates expressing various fusion proteins. Panels of human or animal samples infected with MERS-CoV were used to analyze the sensitivity and specificity of various LISAs in reference to a MERS-CoV RT-PCR, commercial S1-based ELISA, and pseudovirus particle neutralization test (ppNT). Our results showed that the S1-, RBD-, and NP-LISAs were more sensitive than the NTD- and S2-LISAs for the detection of anti-MERS-CoV IgG. Furthermore, the S1-, RBD-, and NP-LISAs were more sensitive (by at least 16-fold) than the commercially available S1-ELISA. Moreover, the S1-, RBD-, and NP-LISA specifically recognized anti-MERS-CoV IgG and did not cross-react with samples derived from other human CoV (OC43, 229E, HKU1, NL63)-infected patients. More importantly, these LISAs proved their applicability and reliability for detecting anti-MERS-CoV IgG in samples from camels, monkeys, and mice, among which the RBD-LISA exhibited excellent performance. The results of this study suggest that the novel MERS-CoV RBD- and S1- LISAs are highly effective platforms for the rapid and sensitive detection of anti-MERS-CoV IgG in human and animal samples. These assays have the potential to be used as serologic tests for the management and control of MERS-CoV infection.
Url:
DOI: 10.1016/j.bsheal.2019.12.006
PubMed: NONE
PubMed Central: 7148641
Affiliations:
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type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Wang, Tianyu" sort="Wang, Tianyu" uniqKey="Wang T" first="Tianyu" last="Wang">Tianyu Wang</name><affiliation wicri:level="1"><nlm:aff id="af0005">MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="1"><nlm:aff id="af0010">Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Epidemiology, School of Public Health, Southern Medical University, Guangzhou 510515, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Epidemiology, School of Public Health, Southern Medical University, Guangzhou 510515</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Deng, Yao" sort="Deng, Yao" uniqKey="Deng Y" first="Yao" last="Deng">Yao Deng</name><affiliation wicri:level="1"><nlm:aff id="af0005">MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Niu, Peihua" sort="Niu, Peihua" uniqKey="Niu P" first="Peihua" last="Niu">Peihua Niu</name><affiliation wicri:level="1"><nlm:aff id="af0005">MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="A, Ruhan" sort="A, Ruhan" uniqKey="A R" first="Ruhan" last="A">Ruhan A</name><affiliation wicri:level="1"><nlm:aff id="af0005">MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></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="af0015">State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Disease, The first Affiliated Hospital of Guangzhou Medical University, Guangzhou 510182, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Disease, The first Affiliated Hospital of Guangzhou Medical University, Guangzhou 510182</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Peiris, Malik" sort="Peiris, Malik" uniqKey="Peiris M" first="Malik" last="Peiris">Malik Peiris</name><affiliation><nlm:aff id="af0020">School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region</nlm:aff><wicri:noCountry code="subfield">Hong Kong Special Administrative Region</wicri:noCountry></affiliation></author><author><name sortKey="Tang, Shixing" sort="Tang, Shixing" uniqKey="Tang S" first="Shixing" last="Tang">Shixing Tang</name><affiliation wicri:level="1"><nlm:aff id="af0010">Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Epidemiology, School of Public Health, Southern Medical University, Guangzhou 510515, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Epidemiology, School of Public Health, Southern Medical University, Guangzhou 510515</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><affiliation wicri:level="1"><nlm:aff id="af0005">MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>MHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 102206</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="1"><nlm:aff id="af0025">Center for Biosafety Mega-science, Chinese Academy of Sciences, Beijing 100101, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Center for Biosafety Mega-science, Chinese Academy of Sciences, Beijing 100101</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author></analytic><series><title level="j">Biosafety and Health</title><idno type="ISSN">2590-0536</idno><idno type="eISSN">2590-0536</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The Middle East respiratory syndrome (MERS) is a lethal zoonosis caused by MERS coronavirus (MERS-CoV) and poses a significant threat to public health worldwide. Therefore, a rapid, sensitive, and specific serologic test for detecting anti-MERS-CoV antibodies in both humans and animals is urgently needed for the successful management of this illness. Here, we evaluated various novel luciferase immunosorbent assays (LISA) based on nucleocapsid protein (NP) as well as fragments derived from spike protein (S) including subunit 1 (S1), N terminal domain (NTD), receptor-binding domain (RBD) and subunit 2 (S2) of S for the detection of MERS-CoV-specific IgG. Fusion proteins, including nanoluciferase (NLuc) and various fragments derived from the NP or S protein of MERS-CoV, were expressed in human embryonic kidney 293 T cells. LISAs that detected anti-MERS-CoV IgG were further developed using cell lysates expressing various fusion proteins. Panels of human or animal samples infected with MERS-CoV were used to analyze the sensitivity and specificity of various LISAs in reference to a MERS-CoV RT-PCR, commercial S1-based ELISA, and pseudovirus particle neutralization test (ppNT). Our results showed that the S1-, RBD-, and NP-LISAs were more sensitive than the NTD- and S2-LISAs for the detection of anti-MERS-CoV IgG. Furthermore, the S1-, RBD-, and NP-LISAs were more sensitive (by at least 16-fold) than the commercially available S1-ELISA. Moreover, the S1-, RBD-, and NP-LISA specifically recognized anti-MERS-CoV IgG and did not cross-react with samples derived from other human CoV (OC43, 229E, HKU1, NL63)-infected patients. More importantly, these LISAs proved their applicability and reliability for detecting anti-MERS-CoV IgG in samples from camels, monkeys, and mice, among which the RBD-LISA exhibited excellent performance. The results of this study suggest that the novel MERS-CoV RBD- and S1- LISAs are highly effective platforms for the rapid and sensitive detection of anti-MERS-CoV IgG in human and animal samples. These assays have the potential to be used as serologic tests for the management and control of MERS-CoV infection.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Al Tayib, O A" uniqKey="Al Tayib O">O.A. Al-Tayib</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gossner, C" uniqKey="Gossner C">C. Gossner</name></author><author><name sortKey="Danielson, N" uniqKey="Danielson N">N. Danielson</name></author><author><name sortKey="Gervelmeyer, A" uniqKey="Gervelmeyer A">A. Gervelmeyer</name></author><author><name sortKey="Berthe, F" uniqKey="Berthe F">F. Berthe</name></author><author><name sortKey="Faye, B" uniqKey="Faye B">B. Faye</name></author><author><name sortKey="Aaslav, K K" uniqKey="Aaslav K">K.K. 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Rahman</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Guangdong</li></region><settlement><li>Jiangmen</li><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Peiris, Malik" sort="Peiris, Malik" uniqKey="Peiris M" first="Malik" last="Peiris">Malik Peiris</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Wenling" sort="Wang, Wenling" uniqKey="Wang W" first="Wenling" last="Wang">Wenling Wang</name></noRegion><name sortKey="A, Ruhan" sort="A, Ruhan" uniqKey="A R" first="Ruhan" last="A">Ruhan A</name><name sortKey="Deng, Yao" sort="Deng, Yao" uniqKey="Deng Y" first="Yao" last="Deng">Yao Deng</name><name sortKey="Niu, Peihua" sort="Niu, Peihua" uniqKey="Niu P" first="Peihua" last="Niu">Peihua Niu</name><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><name sortKey="Tang, Shixing" sort="Tang, Shixing" uniqKey="Tang S" first="Shixing" last="Tang">Shixing Tang</name><name sortKey="Wang, Tianyu" sort="Wang, Tianyu" uniqKey="Wang T" first="Tianyu" last="Wang">Tianyu Wang</name><name sortKey="Wang, Tianyu" sort="Wang, Tianyu" uniqKey="Wang T" first="Tianyu" last="Wang">Tianyu Wang</name><name sortKey="Zhao, Jincun" sort="Zhao, Jincun" uniqKey="Zhao J" first="Jincun" last="Zhao">Jincun Zhao</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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