Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19
Identifieur interne : 004062 ( Main/Exploration ); précédent : 004061; suivant : 004063Synthetic oligonucleotide cocktails as probes for detection of human parvovirus B19
Auteurs : Heather A. Cubie [Royaume-Uni] ; John Grzybowski [Royaume-Uni] ; Chandu Da Silva [Royaume-Uni] ; Linda Duncan [Royaume-Uni] ; Tom Brown [Royaume-Uni] ; Nicholas M. Smith [Royaume-Uni]Source :
- 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
Affiliations:
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Cubie</name></author><author><name sortKey="Grzybowski, John" sort="Grzybowski, John" uniqKey="Grzybowski J" first="John" last="Grzybowski">John Grzybowski</name></author><author><name sortKey="Da Silva, Chandu" sort="Da Silva, Chandu" uniqKey="Da Silva C" first="Chandu" last="Da Silva">Chandu Da Silva</name></author><author><name sortKey="Duncan, Linda" sort="Duncan, Linda" uniqKey="Duncan L" first="Linda" last="Duncan">Linda Duncan</name></author><author><name sortKey="Brown, Tom" sort="Brown, Tom" uniqKey="Brown T" first="Tom" last="Brown">Tom Brown</name></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 wicri:level="2"><country>Royaume-Uni</country><placeName><region type="country">Écosse</region></placeName><wicri:cityArea>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB</wicri:cityArea></affiliation></author><author><name sortKey="Grzybowski, John" sort="Grzybowski, John" uniqKey="Grzybowski J" first="John" last="Grzybowski">John Grzybowski</name><affiliation wicri:level="2"><country>Royaume-Uni</country><placeName><region type="country">Écosse</region></placeName><wicri:cityArea>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ</wicri:cityArea></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 wicri:level="2"><country>Royaume-Uni</country><placeName><region type="country">Écosse</region></placeName><wicri:cityArea>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF</wicri:cityArea></affiliation></author><author><name sortKey="Duncan, Linda" sort="Duncan, Linda" uniqKey="Duncan L" first="Linda" last="Duncan">Linda Duncan</name><affiliation wicri:level="2"><country>Royaume-Uni</country><placeName><region type="country">Écosse</region></placeName><wicri:cityArea>Regional Virus Laboratory, City Hospital, Greenbank Drive, Edinburgh EH10 5SB</wicri:cityArea></affiliation></author><author><name sortKey="Brown, Tom" sort="Brown, Tom" uniqKey="Brown T" first="Tom" last="Brown">Tom Brown</name><affiliation wicri:level="2"><country>Royaume-Uni</country><placeName><region type="country">Écosse</region></placeName><wicri:cityArea>Department of Chemistry, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ</wicri:cityArea></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 wicri:level="2"><country>Royaume-Uni</country><placeName><region type="country">Écosse</region></placeName><wicri:cityArea>Department of Paediatric Pathology, Royal Hospital for Sick Children, Rillbank Terrace, Edinburgh EH9 1LF</wicri:cityArea></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><affiliations><list><country><li>Royaume-Uni</li></country><region><li>Écosse</li></region></list><tree><country name="Royaume-Uni"><region name="Écosse"><name sortKey="Cubie, Heather A" sort="Cubie, Heather A" uniqKey="Cubie H" first="Heather A." last="Cubie">Heather A. Cubie</name></region><name sortKey="Brown, Tom" sort="Brown, Tom" uniqKey="Brown T" first="Tom" last="Brown">Tom Brown</name><name sortKey="Da Silva, Chandu" sort="Da Silva, Chandu" uniqKey="Da Silva C" first="Chandu" last="Da Silva">Chandu Da Silva</name><name sortKey="Duncan, Linda" sort="Duncan, Linda" uniqKey="Duncan L" first="Linda" last="Duncan">Linda Duncan</name><name sortKey="Grzybowski, John" sort="Grzybowski, John" uniqKey="Grzybowski J" first="John" last="Grzybowski">John Grzybowski</name><name sortKey="Smith, Nicholas M" sort="Smith, Nicholas M" uniqKey="Smith N" first="Nicholas M." last="Smith">Nicholas M. Smith</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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