Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19
Identifieur interne : 000510 ( Istex/Corpus ); précédent : 000509; suivant : 000511Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19
Auteurs : Heather A. Cubie ; John Grzybowski ; Chandu Da Silva ; Linda Duncan ; Tom Brown ; Nicholas M. SmithSource :
- Journal of Virological Methods [ 0166-0934 ] ; 1995.
English descriptors
- Teeft :
- Alkaline phosphatase, Amersham hybond membrane, Assay, Blot assay, Blot hybridisation assay, Boehringer mannheim, Cellular specimens, Chemical labelling, Clinical specimens, Consecutive cases, Cubie, Cytospin preparations, Diagnostic applications, Digoxigenin, Edinburgh, Enzymic labelling, Fetal, Fetal hydrops, Final concentration, Greater sensitivity, Health laboratory, High stringency washes, Histological, Histological sections, Human papilloma virus, Human parvovirus, Hybridisation, Hybridisation buffer, Hybridisation temperature, Hydrops, Hydrops fetalis, Hypoplastic aorta, Labelling, Large quantities, Negative cases, Normal karyotype, Nucleic acid, Nylon membrane, Oligonucleotide, Oligonucleotides, Parvovirus, Positive cases, Positive cells, Probe, Probe cocktail, Probe cocktails, Probe concentrations, Room temperature, Single molecule, Single oligonucleotides, Small quantities, Stronger signals, Synthetic oligonucleotides, Virological, Virological methods, Virus reference division, Water bath.
Abstract
Abstract: A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.
Url:
DOI: 10.1016/0166-0934(94)00179-K
Links to Exploration step
ISTEX:ED8B15BFA8DB6C3D6184F820EEACFE714F0EE8BFLe document en format XML
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Cubie</name><affiliation><mods:affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Grzybowski, John" sort="Grzybowski, John" uniqKey="Grzybowski J" first="John" last="Grzybowski">John Grzybowski</name><affiliation><mods:affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Da Silva, Chandu" sort="Da Silva, Chandu" uniqKey="Da Silva C" first="Chandu" last="Da Silva">Chandu Da Silva</name><affiliation><mods:affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Duncan, Linda" sort="Duncan, Linda" uniqKey="Duncan L" first="Linda" last="Duncan">Linda Duncan</name><affiliation><mods:affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Brown, Tom" sort="Brown, Tom" uniqKey="Brown T" first="Tom" last="Brown">Tom Brown</name><affiliation><mods:affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Smith, Nicholas M" sort="Smith, Nicholas M" uniqKey="Smith N" first="Nicholas M." last="Smith">Nicholas M. 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Cubie</name><affiliation><mods:affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Grzybowski, John" sort="Grzybowski, John" uniqKey="Grzybowski J" first="John" last="Grzybowski">John Grzybowski</name><affiliation><mods:affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Da Silva, Chandu" sort="Da Silva, Chandu" uniqKey="Da Silva C" first="Chandu" last="Da Silva">Chandu Da Silva</name><affiliation><mods:affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Duncan, Linda" sort="Duncan, Linda" uniqKey="Duncan L" first="Linda" last="Duncan">Linda Duncan</name><affiliation><mods:affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Brown, Tom" sort="Brown, Tom" uniqKey="Brown T" first="Tom" last="Brown">Tom Brown</name><affiliation><mods:affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</mods:affiliation></affiliation></author><author><name sortKey="Smith, Nicholas M" sort="Smith, Nicholas M" uniqKey="Smith N" first="Nicholas M." last="Smith">Nicholas M. Smith</name><affiliation><mods:affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="ISSN">0166-0934</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1995">1995</date><biblScope unit="volume">53</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="91">91</biblScope><biblScope unit="page" to="102">102</biblScope></imprint><idno type="ISSN">0166-0934</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Alkaline phosphatase</term><term>Amersham hybond membrane</term><term>Assay</term><term>Blot assay</term><term>Blot hybridisation assay</term><term>Boehringer mannheim</term><term>Cellular specimens</term><term>Chemical labelling</term><term>Clinical specimens</term><term>Consecutive cases</term><term>Cubie</term><term>Cytospin preparations</term><term>Diagnostic applications</term><term>Digoxigenin</term><term>Edinburgh</term><term>Enzymic labelling</term><term>Fetal</term><term>Fetal hydrops</term><term>Final concentration</term><term>Greater sensitivity</term><term>Health laboratory</term><term>High stringency washes</term><term>Histological</term><term>Histological sections</term><term>Human papilloma virus</term><term>Human parvovirus</term><term>Hybridisation</term><term>Hybridisation buffer</term><term>Hybridisation temperature</term><term>Hydrops</term><term>Hydrops fetalis</term><term>Hypoplastic aorta</term><term>Labelling</term><term>Large quantities</term><term>Negative cases</term><term>Normal karyotype</term><term>Nucleic acid</term><term>Nylon membrane</term><term>Oligonucleotide</term><term>Oligonucleotides</term><term>Parvovirus</term><term>Positive cases</term><term>Positive cells</term><term>Probe</term><term>Probe cocktail</term><term>Probe cocktails</term><term>Probe concentrations</term><term>Room temperature</term><term>Single molecule</term><term>Single oligonucleotides</term><term>Small quantities</term><term>Stronger signals</term><term>Synthetic oligonucleotides</term><term>Virological</term><term>Virological methods</term><term>Virus reference division</term><term>Water bath</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>hybridisation</json:string><json:string>parvovirus</json:string><json:string>cubie</json:string><json:string>oligonucleotides</json:string><json:string>digoxigenin</json:string><json:string>hydrops</json:string><json:string>human parvovirus</json:string><json:string>fetal</json:string><json:string>oligonucleotide</json:string><json:string>virological</json:string><json:string>virological methods</json:string><json:string>labelling</json:string><json:string>fetal hydrops</json:string><json:string>histological</json:string><json:string>edinburgh</json:string><json:string>probe cocktail</json:string><json:string>histological sections</json:string><json:string>boehringer mannheim</json:string><json:string>synthetic oligonucleotides</json:string><json:string>blot assay</json:string><json:string>assay</json:string><json:string>probe</json:string><json:string>room temperature</json:string><json:string>probe cocktails</json:string><json:string>human papilloma virus</json:string><json:string>nylon membrane</json:string><json:string>consecutive cases</json:string><json:string>single molecule</json:string><json:string>blot hybridisation assay</json:string><json:string>chemical labelling</json:string><json:string>cytospin preparations</json:string><json:string>cellular specimens</json:string><json:string>large quantities</json:string><json:string>virus reference division</json:string><json:string>health laboratory</json:string><json:string>positive cells</json:string><json:string>hybridisation temperature</json:string><json:string>hybridisation buffer</json:string><json:string>water bath</json:string><json:string>nucleic acid</json:string><json:string>alkaline phosphatase</json:string><json:string>stronger signals</json:string><json:string>enzymic labelling</json:string><json:string>final concentration</json:string><json:string>probe concentrations</json:string><json:string>amersham hybond membrane</json:string><json:string>high stringency washes</json:string><json:string>negative cases</json:string><json:string>positive cases</json:string><json:string>hypoplastic aorta</json:string><json:string>normal karyotype</json:string><json:string>single oligonucleotides</json:string><json:string>small quantities</json:string><json:string>greater sensitivity</json:string><json:string>hydrops fetalis</json:string><json:string>clinical specimens</json:string><json:string>diagnostic applications</json:string></teeft></keywords><author><json:item><name>Heather A. Cubie</name><affiliations><json:string>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</json:string></affiliations></json:item><json:item><name>John Grzybowski</name><affiliations><json:string>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</json:string></affiliations></json:item><json:item><name>Chandu Da Silva</name><affiliations><json:string>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</json:string></affiliations></json:item><json:item><name>linda Duncan</name><affiliations><json:string>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</json:string></affiliations></json:item><json:item><name>Tom Brown</name><affiliations><json:string>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</json:string></affiliations></json:item><json:item><name>Nicholas M. Smith</name><affiliations><json:string>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Oligonucleotides</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>ISH</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Parvovirus B19</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>DIG labelling</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>DNP labelling</value></json:item></subject><arkIstex>ark:/67375/6H6-KF8F7TF7-J</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.</abstract><qualityIndicators><score>7.56</score><pdfWordCount>3856</pdfWordCount><pdfCharCount>24186</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>12</pdfPageCount><pdfPageSize>468 x 684 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>142</abstractWordCount><abstractCharCount>944</abstractCharCount><keywordCount>5</keywordCount></qualityIndicators><title>Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19</title><pmid><json:string>7635929</json:string></pmid><pii><json:string>0166-0934(94)00179-K</json:string></pii><genre><json:string>research-article</json:string></genre><host><title>Journal of Virological Methods</title><language><json:string>unknown</json:string></language><publicationDate>1995</publicationDate><issn><json:string>0166-0934</json:string></issn><pii><json:string>S0166-0934(00)X0014-6</json:string></pii><volume>53</volume><issue>1</issue><pages><first>91</first><last>102</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1993</json:string><json:string>1995</json:string><json:string>1984</json:string></date><geogName></geogName><orgName><json:string>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH</json:string><json:string>University of Edinburgh</json:string><json:string>Central Public Health Laboratory, London</json:string><json:string>Boehringer Mannheim UK Ltd</json:string><json:string>Department of Chemistry, University of Edinburgh, King</json:string><json:string>Cruachem Ltd., Glasgow</json:string><json:string>Dako Ltd</json:string><json:string>Dako Ltd.</json:string><json:string>Royal Society of Edinburgh for the Caledonian Research Fellowship</json:string><json:string>Scottish Cot Death Trust</json:string></orgName><orgName_funder><json:string>Royal Society of Edinburgh for the Caledonian Research Fellowship</json:string><json:string>Scottish Cot Death Trust</json:string></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Bernard Cohen</json:string><json:string>B.J. Cohen</json:string><json:string>Nil Nil</json:string><json:string>Pamela Munro</json:string><json:string>Nicholas M. Smith</json:string><json:string>H.A. Cubie</json:string><json:string>Nil Moderate</json:string></persName><placeName><json:string>Mannheim</json:string><json:string>Edinburgh</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Baccard-Longere et al., 1994</json:string><json:string>Cubic and Norval, 1988</json:string><json:string>Cubie and Norval (1988)</json:string><json:string>Mori et al. (1989)</json:string><json:string>Porter et al., 1988</json:string><json:string>Keeling, 1993</json:string><json:string>Cubie et al.</json:string><json:string>Hall, 1994</json:string><json:string>Salimans et al., 1989</json:string><json:string>Morey et al. (1992)</json:string><json:string>February and October, 1993</json:string><json:string>Jasani et al., 1983</json:string><json:string>Cubie and Norval, 1988</json:string><json:string>Sano et al., 1988</json:string><json:string>Cubie et al., 1994</json:string><json:string>Cubie et al., 1991a,b</json:string><json:string>Grzybowski et al., 1993</json:string><json:string>Brown et al., 1984</json:string><json:string>Shade et al., 1986</json:string><json:string>Hewlins et al., 1994</json:string><json:string>Grzybowski et al. (1993)</json:string><json:string>Morey et al., 1992</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-KF8F7TF7-J</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - virology</json:string><json:string>2 - biotechnology & applied microbiology</json:string><json:string>2 - biochemical research methods</json:string></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - virology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Immunology and Microbiology</json:string><json:string>3 - Virology</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences medicales</json:string></inist></categories><publicationDate>1995</publicationDate><copyrightDate>1995</copyrightDate><doi><json:string>10.1016/0166-0934(94)00179-K</json:string></doi><id>ED8B15BFA8DB6C3D6184F820EEACFE714F0EE8BF</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-KF8F7TF7-J/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-KF8F7TF7-J/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/6H6-KF8F7TF7-J/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a">Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>elsevier</p></licence></availability><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M"></p><date>1995</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note type="content">Section title: Research paper</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19</title><author xml:id="author-0000"><persName><forename type="first">Heather A.</forename><surname>Cubie</surname></persName><note type="correspondence"><p>Corresponding author. Tel. 031 536 6221 ext. 3221, Fax 031 536 6153.</p></note><affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</affiliation></author><author xml:id="author-0001"><persName><forename type="first">John</forename><surname>Grzybowski</surname></persName><affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Chandu</forename><surname>Da Silva</surname></persName><affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</affiliation></author><author xml:id="author-0003"><persName><forename type="first">linda</forename><surname>Duncan</surname></persName><affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</affiliation></author><author xml:id="author-0004"><persName><forename type="first">Tom</forename><surname>Brown</surname></persName><affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</affiliation></author><author xml:id="author-0005"><persName><forename type="first">Nicholas M.</forename><surname>Smith</surname></persName><affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</affiliation></author><idno type="istex">ED8B15BFA8DB6C3D6184F820EEACFE714F0EE8BF</idno><idno type="ark">ark:/67375/6H6-KF8F7TF7-J</idno><idno type="DOI">10.1016/0166-0934(94)00179-K</idno><idno type="PII">0166-0934(94)00179-K</idno></analytic><monogr><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="pISSN">0166-0934</idno><idno type="PII">S0166-0934(00)X0014-6</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1995"></date><biblScope unit="volume">53</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="91">91</biblScope><biblScope unit="page" to="102">102</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1995</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>Oligonucleotides</term></item><item><term>ISH</term></item><item><term>Parvovirus B19</term></item><item><term>DIG labelling</term></item><item><term>DNP labelling</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1995">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-KF8F7TF7-J/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>VIRMET</jid><aid>9400179K</aid><ce:pii>0166-0934(94)00179-K</ce:pii><ce:doi>10.1016/0166-0934(94)00179-K</ce:doi><ce:copyright type="unknown" year="1995"></ce:copyright></item-info><head><ce:dochead><ce:textfn>Research paper</ce:textfn></ce:dochead><ce:title>Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19</ce:title><ce:author-group><ce:author><ce:given-name>Heather A.</ce:given-name><ce:surname>Cubie</ce:surname><ce:cross-ref refid="COR1"><ce:sup>∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>John</ce:given-name><ce:surname>Grzybowski</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Chandu</ce:given-name><ce:surname>Da Silva</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>linda</ce:given-name><ce:surname>Duncan</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Tom</ce:given-name><ce:surname>Brown</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Nicholas M.</ce:given-name><ce:surname>Smith</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>c</ce:label><ce:textfn>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>∗</ce:label><ce:text>Corresponding author. Tel. 031 536 6221 ext. 3221, Fax 031 536 6153.</ce:text></ce:correspondence></ce:author-group><ce:date-accepted day="12" month="12" year="1994"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection.</ce:simple-para><ce:simple-para>Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Oligonucleotides</ce:text></ce:keyword><ce:keyword><ce:text>ISH</ce:text></ce:keyword><ce:keyword><ce:text>Parvovirus B19</ce:text></ce:keyword><ce:keyword><ce:text>DIG labelling</ce:text></ce:keyword><ce:keyword><ce:text>DNP labelling</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19</title></titleInfo><name type="personal"><namePart type="given">Heather A.</namePart><namePart type="family">Cubie</namePart><affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</affiliation><description>Corresponding author. Tel. 031 536 6221 ext. 3221, Fax 031 536 6153.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">John</namePart><namePart type="family">Grzybowski</namePart><affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Chandu</namePart><namePart type="family">Da Silva</namePart><affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">linda</namePart><namePart type="family">Duncan</namePart><affiliation>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB, Scotland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Tom</namePart><namePart type="family">Brown</namePart><affiliation>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Nicholas M.</namePart><namePart type="family">Smith</namePart><affiliation>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF, Scotland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1995</dateIssued><copyrightDate encoding="w3cdtf">1995</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: A cocktail of 10 oligonucleotides selected at intervals along the length of the genome of human parvovirus B19 was labelled enzymically with digoxigenin and chemically with either digoxigenin (DIG) or dinitrophenyl (DNP). Chemical labelling was easier and more practical for the production of large quantities of probe. Pools labelled with either digoxigenin or DNP could detect 10 fg of B19 DNA in a dot blot reaction using an alkaline phosphatase antibody conjugate and colorimetric detection. Formalin fixed tissue from 11 consecutive cases of fetal hydrops were examined by in situ hybridisation (ISH). Both probe cocktails detected human parvovirus B19 DNA in 3 cases, with positive cells in all tissues examined and with equal sensitivity. The DNP pool is significantly cheaper and simpler to produce and could provide an inexpensive reagent suitable for diagnostic detection of viral nucleic acid in histopathological material.</abstract><note type="content">Section title: Research paper</note><subject><genre>Keywords</genre><topic>Oligonucleotides</topic><topic>ISH</topic><topic>Parvovirus B19</topic><topic>DIG labelling</topic><topic>DNP labelling</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Virological Methods</title></titleInfo><titleInfo type="abbreviated"><title>VIRMET</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1995</dateIssued></originInfo><identifier type="ISSN">0166-0934</identifier><identifier type="PII">S0166-0934(00)X0014-6</identifier><part><date>1995</date><detail type="volume"><number>53</number><caption>vol.</caption></detail><detail type="issue"><number>1</number><caption>no.</caption></detail><extent unit="issue-pages"><start>1</start><end>166</end></extent><extent unit="pages"><start>91</start><end>102</end></extent></part></relatedItem><identifier type="istex">ED8B15BFA8DB6C3D6184F820EEACFE714F0EE8BF</identifier><identifier type="ark">ark:/67375/6H6-KF8F7TF7-J</identifier><identifier type="DOI">10.1016/0166-0934(94)00179-K</identifier><identifier type="PII">0166-0934(94)00179-K</identifier><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-KF8F7TF7-J/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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