Application of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum
Infection with African swine fever virus (ASFV) causes an acute and highly lethal hemorrhagic disease that has been responsible for huge economic losses in China. To exactly detect the antigen of ASFV, we established a rapid, no-wash, one-step sandwich-type immunoassay based on the amplified lumines...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2021-06, Vol.105 (11), p.4751-4759 |
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| creator | Chen, Dongjie Wang, Di Wang, Caixia Wei, Fang Zhao, Hongyuan Lin, Xiangmei Wu, Shaoqiang |
| description | Infection with African swine fever virus (ASFV) causes an acute and highly lethal hemorrhagic disease that has been responsible for huge economic losses in China. To exactly detect the antigen of ASFV, we established a rapid, no-wash, one-step sandwich-type immunoassay based on the amplified luminescent proximity homogenous assay-linked immunosorbent assay (AlphaLISA) using two monoclonal antibodies (mAbs) M-5 and M-6 against ASFV p72. ASFV p72 in samples was captured by biotinylated mAb M-5 connected to the donor bead surface via streptavidin and “sandwiched” by mAb M-6 which was coated onto the acceptor bead. Efficacy and sensitivity trials revealed that the AlphaLISA could detect ≥0.78 ng/ml of purified p72 and with a linear range of 0.78–100 ng/ml. The AlphaLISA was specific for ASFV and did not cross-react with other common pathogenic porcine viruses. Compared with RealPCR ASFV DNA test and ASFV antigen detection kit, the sensitivity of the AlphaLISA evaluated in 60 porcine serum samples was 93% and 100%, respectively. The specificity was 100% and 91.7%, respectively. This study presents a good laboratory diagnostic tool for sensitive and efficient detection of ASFV in porcine serum.
Key points
•
MAbs M-5 and M-6 recognized various epitopes of ASFV p72.
•
The established ASFV p72 AlphaLISA showed well specificity, high sensitivity, and satisfied correlation coefficient. |
| doi_str_mv | 10.1007/s00253-021-11339-2 |
| format | Article |
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Key points
•
MAbs M-5 and M-6 recognized various epitopes of ASFV p72.
•
The established ASFV p72 AlphaLISA showed well specificity, high sensitivity, and satisfied correlation coefficient.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-021-11339-2</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>African swine fever ; Antigens ; Asfarviridae ; Biomedical and Life Sciences ; Biotechnology ; Correlation coefficient ; Correlation coefficients ; Economic impact ; Enzyme-linked immunosorbent assay ; Epitopes ; Fever ; Genetic aspects ; Hemorrhage ; Hemorrhagic disease ; Immunoassay ; Life Sciences ; Methods ; Methods and Protocols ; Microbial Genetics and Genomics ; Microbiology ; Monoclonal antibodies ; Sensitivity analysis ; Streptavidin ; Swine ; Viruses</subject><ispartof>Applied microbiology and biotechnology, 2021-06, Vol.105 (11), p.4751-4759</ispartof><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021</rights><rights>COPYRIGHT 2021 Springer</rights><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c490t-91772ace291670bbd73de1d123c089ea560f4cb6b6e06c1306f05ae6c0ba8913</citedby><cites>FETCH-LOGICAL-c490t-91772ace291670bbd73de1d123c089ea560f4cb6b6e06c1306f05ae6c0ba8913</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-021-11339-2$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-021-11339-2$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids></links><search><creatorcontrib>Chen, Dongjie</creatorcontrib><creatorcontrib>Wang, Di</creatorcontrib><creatorcontrib>Wang, Caixia</creatorcontrib><creatorcontrib>Wei, Fang</creatorcontrib><creatorcontrib>Zhao, Hongyuan</creatorcontrib><creatorcontrib>Lin, Xiangmei</creatorcontrib><creatorcontrib>Wu, Shaoqiang</creatorcontrib><title>Application of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>Infection with African swine fever virus (ASFV) causes an acute and highly lethal hemorrhagic disease that has been responsible for huge economic losses in China. To exactly detect the antigen of ASFV, we established a rapid, no-wash, one-step sandwich-type immunoassay based on the amplified luminescent proximity homogenous assay-linked immunosorbent assay (AlphaLISA) using two monoclonal antibodies (mAbs) M-5 and M-6 against ASFV p72. ASFV p72 in samples was captured by biotinylated mAb M-5 connected to the donor bead surface via streptavidin and “sandwiched” by mAb M-6 which was coated onto the acceptor bead. Efficacy and sensitivity trials revealed that the AlphaLISA could detect ≥0.78 ng/ml of purified p72 and with a linear range of 0.78–100 ng/ml. The AlphaLISA was specific for ASFV and did not cross-react with other common pathogenic porcine viruses. Compared with RealPCR ASFV DNA test and ASFV antigen detection kit, the sensitivity of the AlphaLISA evaluated in 60 porcine serum samples was 93% and 100%, respectively. The specificity was 100% and 91.7%, respectively. This study presents a good laboratory diagnostic tool for sensitive and efficient detection of ASFV in porcine serum.
Key points
•
MAbs M-5 and M-6 recognized various epitopes of ASFV p72.
•
The established ASFV p72 AlphaLISA showed well specificity, high sensitivity, and satisfied correlation coefficient.</description><subject>African swine fever</subject><subject>Antigens</subject><subject>Asfarviridae</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Correlation coefficient</subject><subject>Correlation coefficients</subject><subject>Economic impact</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Epitopes</subject><subject>Fever</subject><subject>Genetic aspects</subject><subject>Hemorrhage</subject><subject>Hemorrhagic disease</subject><subject>Immunoassay</subject><subject>Life Sciences</subject><subject>Methods</subject><subject>Methods and Protocols</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Monoclonal antibodies</subject><subject>Sensitivity analysis</subject><subject>Streptavidin</subject><subject>Swine</subject><subject>Viruses</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kl2L1DAUhosoOK7-Aa8C3uhF15O0TdrLsvgxMCC4ex_S9GQ2S5vUJB3135vZ2WUZEclF4PA8LznhLYq3FC4pgPgYAVhTlcBoSWlVdSV7VmxoXbESOK2fFxugoilF07Uvi1cx3gFQ1nK-KVy_LJPVKlnviDdEOdJPy63aba97MmO69SMxPpCgFjuSiC7aZA9IRkyoH6XehBzhSPxpHRKDBwzkYMMaiXVk8UEfxxHDOr8uXhg1RXzzcF8UN58_3Vx9LXffvmyv-l2p6w5S2VEhmNLIOsoFDMMoqhHpSFmloe1QNRxMrQc-cASuaQXcQKOQaxhU29Hqonh_il2C_7FiTHK2UeM0KYd-jTL_Vc0pg3v03V_onV-Dy4_LVA0AjajFE7VXE0rrjE9B6WOo7DnnrAPedpm6_AeVz4iz1d6hsXl-Jnw4EzKT8FfaqzVGub3-fs6yE6uDjzGgkUuwswq_JQV5LIE8lUDmEsj7EkiWpeokxQy7PYan7f5j_QF82bHv</recordid><startdate>20210601</startdate><enddate>20210601</enddate><creator>Chen, 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of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum</title><author>Chen, Dongjie ; Wang, Di ; Wang, Caixia ; Wei, Fang ; Zhao, Hongyuan ; Lin, Xiangmei ; Wu, Shaoqiang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c490t-91772ace291670bbd73de1d123c089ea560f4cb6b6e06c1306f05ae6c0ba8913</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>African swine fever</topic><topic>Antigens</topic><topic>Asfarviridae</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Correlation coefficient</topic><topic>Correlation coefficients</topic><topic>Economic impact</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Epitopes</topic><topic>Fever</topic><topic>Genetic aspects</topic><topic>Hemorrhage</topic><topic>Hemorrhagic disease</topic><topic>Immunoassay</topic><topic>Life 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biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Dongjie</au><au>Wang, Di</au><au>Wang, Caixia</au><au>Wei, Fang</au><au>Zhao, Hongyuan</au><au>Lin, Xiangmei</au><au>Wu, Shaoqiang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Application of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><date>2021-06-01</date><risdate>2021</risdate><volume>105</volume><issue>11</issue><spage>4751</spage><epage>4759</epage><pages>4751-4759</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>Infection with African swine fever virus (ASFV) causes an acute and highly lethal hemorrhagic disease that has been responsible for huge economic losses in China. To exactly detect the antigen of ASFV, we established a rapid, no-wash, one-step sandwich-type immunoassay based on the amplified luminescent proximity homogenous assay-linked immunosorbent assay (AlphaLISA) using two monoclonal antibodies (mAbs) M-5 and M-6 against ASFV p72. ASFV p72 in samples was captured by biotinylated mAb M-5 connected to the donor bead surface via streptavidin and “sandwiched” by mAb M-6 which was coated onto the acceptor bead. Efficacy and sensitivity trials revealed that the AlphaLISA could detect ≥0.78 ng/ml of purified p72 and with a linear range of 0.78–100 ng/ml. The AlphaLISA was specific for ASFV and did not cross-react with other common pathogenic porcine viruses. Compared with RealPCR ASFV DNA test and ASFV antigen detection kit, the sensitivity of the AlphaLISA evaluated in 60 porcine serum samples was 93% and 100%, respectively. The specificity was 100% and 91.7%, respectively. This study presents a good laboratory diagnostic tool for sensitive and efficient detection of ASFV in porcine serum.
Key points
•
MAbs M-5 and M-6 recognized various epitopes of ASFV p72.
•
The established ASFV p72 AlphaLISA showed well specificity, high sensitivity, and satisfied correlation coefficient.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><doi>10.1007/s00253-021-11339-2</doi><tpages>9</tpages></addata></record> |
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| subjects | African swine fever Antigens Asfarviridae Biomedical and Life Sciences Biotechnology Correlation coefficient Correlation coefficients Economic impact Enzyme-linked immunosorbent assay Epitopes Fever Genetic aspects Hemorrhage Hemorrhagic disease Immunoassay Life Sciences Methods Methods and Protocols Microbial Genetics and Genomics Microbiology Monoclonal antibodies Sensitivity analysis Streptavidin Swine Viruses |
| title | Application of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum |
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