Quantification of infectious duck hepatitis B virus by radioimmunofocus assay
A simple method is described for the precise quantification of infectious duck hepatitis B virus (DHBV) in cell culture, using a radioimmunofocus assay (RIFA). Primary duck hepatocyte cell cultures were infected with serial dilutions of viral samples as for a plaque assay, but then maintained with l...
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| Veröffentlicht in: | Journal of medical virology 1997-08, Vol.52 (4), p.354-361 |
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| container_title | Journal of medical virology |
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| creator | Anderson, David A. Grgacic, Elizabeth V. L. Luscombe, Carolyn A. Gu, Xingnian Dixon, Robert |
| description | A simple method is described for the precise quantification of infectious duck hepatitis B virus (DHBV) in cell culture, using a radioimmunofocus assay (RIFA). Primary duck hepatocyte cell cultures were infected with serial dilutions of viral samples as for a plaque assay, but then maintained with liquid overlay medium. After incubation for up to 14 days, cell monolayers were fixed with acetone, then stained with a monoclonal antibody to DHBV L protein followed by secondary antibody labelled with I. Foci of infection (representing individual infectious particles in the inoculum) were detected by autoradiography. The number of foci recovered was increased by addition of dimethyl sulphoxide to culture medium, but was not appreciably altered by the use of semi‐solid medium. The titre of virus suspensions determined by RIFA correlated well with titration in ducklings. The RIFA is a useful method for titration of DHBV, as it has a wide dynamic range and is well suited to parallel titration of large numbers of samples. This assay will have wide use for the analysis of DHBV growth kinetics, antiviral efficacy, and virus inactivation procedures. J. Med. Virol. 52:354–361, 1997. © 1997 Wiley‐Liss, Inc. |
| doi_str_mv | 10.1002/(SICI)1096-9071(199708)52:4<354::AID-JMV2>3.0.CO;2-0 |
| format | Article |
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The titre of virus suspensions determined by RIFA correlated well with titration in ducklings. The RIFA is a useful method for titration of DHBV, as it has a wide dynamic range and is well suited to parallel titration of large numbers of samples. This assay will have wide use for the analysis of DHBV growth kinetics, antiviral efficacy, and virus inactivation procedures. J. Med. Virol. 52:354–361, 1997. © 1997 Wiley‐Liss, Inc.</description><identifier>ISSN: 0146-6615</identifier><identifier>EISSN: 1096-9071</identifier><identifier>DOI: 10.1002/(SICI)1096-9071(199708)52:4<354::AID-JMV2>3.0.CO;2-0</identifier><identifier>PMID: 9260680</identifier><identifier>CODEN: JMVIDB</identifier><language>eng</language><publisher>New York: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; Antibodies, Monoclonal ; Biological and medical sciences ; Cell Count ; Cells, Cultured ; DHBV ; DNA, Viral - isolation & purification ; Ducks ; Evaluation Studies as Topic ; Fundamental and applied biological sciences. Psychology ; Hepatitis B Virus, Duck - immunology ; Hepatitis B Virus, Duck - isolation & purification ; Hepatitis B Virus, Duck - pathogenicity ; infectivity assay ; Microbiology ; Radioimmunoassay - methods ; Radioimmunoassay - statistics & numerical data ; Sensitivity and Specificity ; Techniques used in virology ; viral quantification ; Virology ; Virology - methods ; Virology - statistics & numerical data</subject><ispartof>Journal of medical virology, 1997-08, Vol.52 (4), p.354-361</ispartof><rights>Copyright © 1997 Wiley‐Liss, Inc.</rights><rights>1997 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c4582-7fba093279d284857e92510d66874e31ac10c2aafc3e77d5f50f854bacfbf72c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2F%28SICI%291096-9071%28199708%2952%3A4%3C354%3A%3AAID-JMV2%3E3.0.CO%3B2-0$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2F%28SICI%291096-9071%28199708%2952%3A4%3C354%3A%3AAID-JMV2%3E3.0.CO%3B2-0$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1416,27922,27923,45572,45573</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2788328$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9260680$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Anderson, David A.</creatorcontrib><creatorcontrib>Grgacic, Elizabeth V. L.</creatorcontrib><creatorcontrib>Luscombe, Carolyn A.</creatorcontrib><creatorcontrib>Gu, Xingnian</creatorcontrib><creatorcontrib>Dixon, Robert</creatorcontrib><title>Quantification of infectious duck hepatitis B virus by radioimmunofocus assay</title><title>Journal of medical virology</title><addtitle>J. Med. Virol</addtitle><description>A simple method is described for the precise quantification of infectious duck hepatitis B virus (DHBV) in cell culture, using a radioimmunofocus assay (RIFA). Primary duck hepatocyte cell cultures were infected with serial dilutions of viral samples as for a plaque assay, but then maintained with liquid overlay medium. After incubation for up to 14 days, cell monolayers were fixed with acetone, then stained with a monoclonal antibody to DHBV L protein followed by secondary antibody labelled with I. Foci of infection (representing individual infectious particles in the inoculum) were detected by autoradiography. The number of foci recovered was increased by addition of dimethyl sulphoxide to culture medium, but was not appreciably altered by the use of semi‐solid medium. The titre of virus suspensions determined by RIFA correlated well with titration in ducklings. The RIFA is a useful method for titration of DHBV, as it has a wide dynamic range and is well suited to parallel titration of large numbers of samples. This assay will have wide use for the analysis of DHBV growth kinetics, antiviral efficacy, and virus inactivation procedures. J. Med. Virol. 52:354–361, 1997. © 1997 Wiley‐Liss, Inc.</description><subject>Animals</subject><subject>Antibodies, Monoclonal</subject><subject>Biological and medical sciences</subject><subject>Cell Count</subject><subject>Cells, Cultured</subject><subject>DHBV</subject><subject>DNA, Viral - isolation & purification</subject><subject>Ducks</subject><subject>Evaluation Studies as Topic</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hepatitis B Virus, Duck - immunology</subject><subject>Hepatitis B Virus, Duck - isolation & purification</subject><subject>Hepatitis B Virus, Duck - pathogenicity</subject><subject>infectivity assay</subject><subject>Microbiology</subject><subject>Radioimmunoassay - methods</subject><subject>Radioimmunoassay - statistics & numerical data</subject><subject>Sensitivity and Specificity</subject><subject>Techniques used in virology</subject><subject>viral quantification</subject><subject>Virology</subject><subject>Virology - methods</subject><subject>Virology - statistics & numerical data</subject><issn>0146-6615</issn><issn>1096-9071</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUdFu0zAUjRBodINPQMoDQttDyrUdx06ZkLYMSlFLhSjweOU6tjBLkxIng_49Dq3KA0h7su49x8fH50TRJYExAaAvzz_NitkFgTxLchDknOS5AHnB6SS9ZDydTK5mN8n7xRf6mo1hXCxf0QQeRKPjhYfRCEiaJVlG-OPo1PvvACBzSk-ik5xmkEkYRYuPvao7Z51WnWvquLGxq63RYeh9XPb6Nv5mtgHrnI-v4zvXhvV6F7eqdI3bbPq6sY0OO-W92j2JHllVefP0cJ5Fn9--WRXvkvlyOiuu5olOuaSJsGsFOaMiL6lMJRcmp5xAmWVSpIYRpQloqpTVzAhRcsvBSp6ulbZrK6hmZ9GLve62bX70xne4cV6bqlK1Cb5RhF-ClOReIskYCMEH4mpP1G3jfWssblu3Ue0OCeBQB-JQBw7p4pAu7utATjHFUAdiqAOHOpAhYLFEihBknx3e79cbUx5FD_kH_PkBV16ryraq1s4faVRIyaj86-6nq8zuH2v3OPuPsT9zkE32ss535tdRVrW3mAkmOH79MMXVfCGmeXqNN-w3PyvAog</recordid><startdate>199708</startdate><enddate>199708</enddate><creator>Anderson, David A.</creator><creator>Grgacic, Elizabeth V. L.</creator><creator>Luscombe, Carolyn A.</creator><creator>Gu, Xingnian</creator><creator>Dixon, Robert</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>199708</creationdate><title>Quantification of infectious duck hepatitis B virus by radioimmunofocus assay</title><author>Anderson, David A. ; Grgacic, Elizabeth V. L. ; Luscombe, Carolyn A. ; Gu, Xingnian ; Dixon, Robert</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4582-7fba093279d284857e92510d66874e31ac10c2aafc3e77d5f50f854bacfbf72c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal</topic><topic>Biological and medical sciences</topic><topic>Cell Count</topic><topic>Cells, Cultured</topic><topic>DHBV</topic><topic>DNA, Viral - isolation & purification</topic><topic>Ducks</topic><topic>Evaluation Studies as Topic</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hepatitis B Virus, Duck - immunology</topic><topic>Hepatitis B Virus, Duck - isolation & purification</topic><topic>Hepatitis B Virus, Duck - pathogenicity</topic><topic>infectivity assay</topic><topic>Microbiology</topic><topic>Radioimmunoassay - methods</topic><topic>Radioimmunoassay - statistics & numerical data</topic><topic>Sensitivity and Specificity</topic><topic>Techniques used in virology</topic><topic>viral quantification</topic><topic>Virology</topic><topic>Virology - methods</topic><topic>Virology - statistics & numerical data</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Anderson, David A.</creatorcontrib><creatorcontrib>Grgacic, Elizabeth V. L.</creatorcontrib><creatorcontrib>Luscombe, Carolyn A.</creatorcontrib><creatorcontrib>Gu, Xingnian</creatorcontrib><creatorcontrib>Dixon, Robert</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of medical virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Anderson, David A.</au><au>Grgacic, Elizabeth V. L.</au><au>Luscombe, Carolyn A.</au><au>Gu, Xingnian</au><au>Dixon, Robert</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of infectious duck hepatitis B virus by radioimmunofocus assay</atitle><jtitle>Journal of medical virology</jtitle><addtitle>J. Med. Virol</addtitle><date>1997-08</date><risdate>1997</risdate><volume>52</volume><issue>4</issue><spage>354</spage><epage>361</epage><pages>354-361</pages><issn>0146-6615</issn><eissn>1096-9071</eissn><coden>JMVIDB</coden><abstract>A simple method is described for the precise quantification of infectious duck hepatitis B virus (DHBV) in cell culture, using a radioimmunofocus assay (RIFA). Primary duck hepatocyte cell cultures were infected with serial dilutions of viral samples as for a plaque assay, but then maintained with liquid overlay medium. After incubation for up to 14 days, cell monolayers were fixed with acetone, then stained with a monoclonal antibody to DHBV L protein followed by secondary antibody labelled with I. Foci of infection (representing individual infectious particles in the inoculum) were detected by autoradiography. The number of foci recovered was increased by addition of dimethyl sulphoxide to culture medium, but was not appreciably altered by the use of semi‐solid medium. The titre of virus suspensions determined by RIFA correlated well with titration in ducklings. The RIFA is a useful method for titration of DHBV, as it has a wide dynamic range and is well suited to parallel titration of large numbers of samples. This assay will have wide use for the analysis of DHBV growth kinetics, antiviral efficacy, and virus inactivation procedures. J. Med. Virol. 52:354–361, 1997. © 1997 Wiley‐Liss, Inc.</abstract><cop>New York</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>9260680</pmid><doi>10.1002/(SICI)1096-9071(199708)52:4<354::AID-JMV2>3.0.CO;2-0</doi><tpages>8</tpages></addata></record> |
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| ispartof | Journal of medical virology, 1997-08, Vol.52 (4), p.354-361 |
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| subjects | Animals Antibodies, Monoclonal Biological and medical sciences Cell Count Cells, Cultured DHBV DNA, Viral - isolation & purification Ducks Evaluation Studies as Topic Fundamental and applied biological sciences. Psychology Hepatitis B Virus, Duck - immunology Hepatitis B Virus, Duck - isolation & purification Hepatitis B Virus, Duck - pathogenicity infectivity assay Microbiology Radioimmunoassay - methods Radioimmunoassay - statistics & numerical data Sensitivity and Specificity Techniques used in virology viral quantification Virology Virology - methods Virology - statistics & numerical data |
| title | Quantification of infectious duck hepatitis B virus by radioimmunofocus assay |
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