Detection of human rhinovirus RNA in nasal washings by PCR
Identifieur interne : 000546 ( Istex/Corpus ); précédent : 000545; suivant : 000547Detection of human rhinovirus RNA in nasal washings by PCR
Auteurs : E. Arruda ; F. G. HaydenSource :
- Molecular and Cellular Probes [ 0890-8508 ] ; 1993.
English descriptors
Abstract
Abstract: A PCR assay was developed to detect human rhinovirus (HRV) RNA in nasal washings from individuals experimentally infected with HRV-39 or HRV strain Hanks. Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID50) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. A GTC-based method for HRV RNA extraction in nasal washings was superior to a proteinase K-phenol/chloroform-based method in regard to sensitivity, consumption of reagents, material and time.
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DOI: 10.1006/mcpr.1993.1055
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Detection of human rhinovirus RNA in nasal washings by PCR</title><author><name sortKey="Arruda, E" sort="Arruda, E" uniqKey="Arruda E" first="E." last="Arruda">E. Arruda</name><affiliation><mods:affiliation>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</mods:affiliation></affiliation></author><author><name sortKey="Hayden, F G" sort="Hayden, F G" uniqKey="Hayden F" first="F. G." last="Hayden">F. G. Hayden</name><affiliation><mods:affiliation>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:A1F5E1A591AAF16BEE479E270C469EDF4AF5751A</idno><date when="1993" year="1993">1993</date><idno type="doi">10.1006/mcpr.1993.1055</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-N1G3R24L-6/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000546</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000546</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Detection of human rhinovirus RNA in nasal washings by PCR</title><author><name sortKey="Arruda, E" sort="Arruda, E" uniqKey="Arruda E" first="E." last="Arruda">E. Arruda</name><affiliation><mods:affiliation>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</mods:affiliation></affiliation></author><author><name sortKey="Hayden, F G" sort="Hayden, F G" uniqKey="Hayden F" first="F. G." last="Hayden">F. G. Hayden</name><affiliation><mods:affiliation>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Molecular and Cellular Probes</title><title level="j" type="abbrev">YMCPR</title><idno type="ISSN">0890-8508</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1993">1993</date><biblScope unit="volume">7</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="373">373</biblScope><biblScope unit="page" to="379">379</biblScope></imprint><idno type="ISSN">0890-8508</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0890-8508</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Rhinovirus, polymerase chain reaction, picornavirus</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: A PCR assay was developed to detect human rhinovirus (HRV) RNA in nasal washings from individuals experimentally infected with HRV-39 or HRV strain Hanks. Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID50) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. A GTC-based method for HRV RNA extraction in nasal washings was superior to a proteinase K-phenol/chloroform-based method in regard to sensitivity, consumption of reagents, material and time.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>E. Arruda</name><affiliations><json:string>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</json:string></affiliations></json:item><json:item><name>F.G. 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Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID50) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. 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Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID50) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. A GTC-based method for HRV RNA extraction in nasal washings was superior to a proteinase K-phenol/chloroform-based method in regard to sensitivity, consumption of reagents, material and time.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>Keywords</head><item><term>Rhinovirus, polymerase chain reaction, picornavirus</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1993">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-N1G3R24L-6/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier converted-article found"><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" xml:lang="en"><item-info><jid>YMCPR</jid><aid>71055</aid><ce:pii>S0890-8508(83)71055-8</ce:pii><ce:doi>10.1006/mcpr.1993.1055</ce:doi><ce:copyright type="full-transfer" year="1993">Academic Press</ce:copyright></item-info><head><ce:dochead><ce:textfn>Original Articles</ce:textfn></ce:dochead><ce:title>Detection of human rhinovirus RNA in nasal washings by PCR</ce:title><ce:author-group><ce:author><ce:given-name>E.</ce:given-name><ce:surname>Arruda</ce:surname></ce:author><ce:author><ce:given-name>F.G.</ce:given-name><ce:surname>Hayden</ce:surname></ce:author><ce:affiliation><ce:textfn>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</ce:textfn></ce:affiliation></ce:author-group><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>A PCR assay was developed to detect human rhinovirus (HRV) RNA in nasal washings from individuals experimentally infected with HRV-39 or HRV strain Hanks. Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID<ce:inf>50</ce:inf>) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. A GTC-based method for HRV RNA extraction in nasal washings was superior to a proteinase K-phenol/chloroform-based method in regard to sensitivity, consumption of reagents, material and time.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Rhinovirus, polymerase chain reaction, picornavirus</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Detection of human rhinovirus RNA in nasal washings by PCR</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Detection of human rhinovirus RNA in nasal washings by PCR</title></titleInfo><name type="personal"><namePart type="given">E.</namePart><namePart type="family">Arruda</namePart><affiliation>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">F.G.</namePart><namePart type="family">Hayden</namePart><affiliation>Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia, USA</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">1993</dateIssued><copyrightDate encoding="w3cdtf">1993</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: A PCR assay was developed to detect human rhinovirus (HRV) RNA in nasal washings from individuals experimentally infected with HRV-39 or HRV strain Hanks. Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID50) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. 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