Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens
Identifieur interne : 004430 ( Main/Exploration ); précédent : 004429; suivant : 004431Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens
Auteurs : A. Petrich [Canada] ; J. Mahony [Canada] ; S. Chong [Canada] ; G. Broukhanski [Canada] ; F. Gharabaghi [Canada] ; G. Johnson [Canada] ; L. Louie [Canada] ; K. Luinstra [Canada] ; B. Willey [Canada] ; P. Akhaven [Canada] ; L. Chui [Canada] ; F. Jamieson [Canada] ; M. Louie [Canada] ; T. Mazzulli [Canada] ; R. Tellier [Canada] ; M. Smieja [Canada] ; W. Cai [Canada] ; M. Chernesky [Canada] ; S. E. Richardson [Canada]Source :
- Journal of clinical microbiology : (Print) [ 0095-1137 ] ; 2006.
Descripteurs français
- KwdFr :
- ARN viral (analyse), ARN viral (génétique), ARN viral (isolement et purification), Fèces (virologie), Humains, RT-PCR, Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Techniques de diagnostic moléculaire, Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- analyse : ARN viral.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ARN viral, Virus du SRAS.
- isolement et purification : ARN viral, Virus du SRAS.
- virologie : Fèces, Syndrome respiratoire aigu sévère.
- Pascal (Inist)
English descriptors
- KwdEn :
- Coronavirus, Detection, Feces, Feces (virology), Humans, Method, Microbiology, Molecular Diagnostic Techniques, Nucleic acid, RNA, Viral (analysis), RNA, Viral (genetics), RNA, Viral (isolation & purification), Reverse Transcriptase Polymerase Chain Reaction, SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Severe acute respiratory syndrome.
- MESH :
- chemical , analysis : RNA, Viral.
- chemical , genetics : RNA, Viral.
- chemical , isolation & purification : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- virology : Feces, Severe Acute Respiratory Syndrome.
- Humans, Molecular Diagnostic Techniques, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity.
Abstract
The emergence of a novel coronavirus (CoV) as the cause of severe acute respiratory syndrome (SARS) catalyzed the development of rapid diagnostic tests. Stool samples have been shown to be appropriate for diagnostic testing for SARS CoV, although it has been recognized to be a heterogeneous and difficult sample that contains amplification inhibitors. Limited information on the efficiency of extraction methods for the purification and concentration of SARS CoV RNA from stool samples is available. Our study objectives were to determine the optimal extraction method for SARS CoV RNA detection and to examine the effect of increased specimen volume for the detection of SARS CoV RNA in stool specimens. We conducted a multicenter evaluation of four automated and four manual extraction methods using dilutions of viral lysate in replicate mock stool samples, followed by quantitation of SARS CoV RNA using real-time reverse transcriptase PCR. The sensitivities of the manual methods ranged from 50% to 100%, with the Cortex Biochem Magazorb method, a magnetic bead isolation method, allowing detection of all 12 positive samples. The sensitivities of the automated methods ranged from 75% to 100%. The bioMérieux NucliSens automated extractor and miniMag extraction methods each had a sensitivity of 100%. Examination of the copy numbers detected and the generation of 10-fold dilutions of the extracted material indicated that a number of extraction methods retained inhibitory substances that prevented optimal amplification. Increasing the volume of sample input did improve detection. This information could be useful for the extraction of other RNA viruses from stool samples and demonstrates the need to evaluate extraction methods for different specimen types.
Affiliations:
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Le document en format XML
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens</title>
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<author><name sortKey="Chong, S" sort="Chong, S" uniqKey="Chong S" first="S." last="Chong">S. Chong</name>
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<author><name sortKey="Broukhanski, G" sort="Broukhanski, G" uniqKey="Broukhanski G" first="G." last="Broukhanski">G. Broukhanski</name>
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<author><name sortKey="Luinstra, K" sort="Luinstra, K" uniqKey="Luinstra K" first="K." last="Luinstra">K. Luinstra</name>
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<s2>Hamilton, Ontario</s2>
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<author><name sortKey="Willey, B" sort="Willey, B" uniqKey="Willey B" first="B." last="Willey">B. Willey</name>
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</affiliation>
</author>
<author><name sortKey="Akhaven, P" sort="Akhaven, P" uniqKey="Akhaven P" first="P." last="Akhaven">P. Akhaven</name>
<affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Mt. Sinai Hospital</s1>
<s2>Toronto, Ontario</s2>
<s3>CAN</s3>
<sZ>9 aut.</sZ>
<sZ>10 aut.</sZ>
<sZ>14 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Mt. Sinai Hospital</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chui, L" sort="Chui, L" uniqKey="Chui L" first="L." last="Chui">L. Chui</name>
<affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Edmonton Public Health Laboratory</s1>
<s2>Edmonton, Alberta</s2>
<s3>CAN</s3>
<sZ>11 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Edmonton Public Health Laboratory</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><inist:fA14 i1="08"><s1>Calgary Public Health Laboratory</s1>
<s2>Calgary, Alberta</s2>
<s3>CAN</s3>
<sZ>11 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Calgary Public Health Laboratory</wicri:noRegion>
</affiliation>
<affiliation wicri:level="4"><inist:fA14 i1="09"><s1>McMaster University</s1>
<s2>Hamilton, Ontario</s2>
<s3>CAN</s3>
<sZ>11 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<placeName><settlement type="city">Hamilton (Ontario)</settlement>
<region type="state">Ontario</region>
</placeName>
<orgName type="university">Université McMaster</orgName>
</affiliation>
</author>
<author><name sortKey="Jamieson, F" sort="Jamieson, F" uniqKey="Jamieson F" first="F." last="Jamieson">F. Jamieson</name>
<affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Ontario Ministry of Health</s1>
<s2>Etobicoke, Ontario</s2>
<s3>CAN</s3>
<sZ>4 aut.</sZ>
<sZ>12 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Ontario Ministry of Health</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Louie, M" sort="Louie, M" uniqKey="Louie M" first="M." last="Louie">M. Louie</name>
<affiliation wicri:level="4"><inist:fA14 i1="10"><s1>The University of Toronto</s1>
<s2>Toronto, Ontario</s2>
<s3>CAN</s3>
<sZ>13 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<placeName><settlement type="city">Toronto</settlement>
<region type="state">Ontario</region>
</placeName>
<orgName type="university">Université de Toronto</orgName>
</affiliation>
</author>
<author><name sortKey="Mazzulli, T" sort="Mazzulli, T" uniqKey="Mazzulli T" first="T." last="Mazzulli">T. Mazzulli</name>
<affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Mt. Sinai Hospital</s1>
<s2>Toronto, Ontario</s2>
<s3>CAN</s3>
<sZ>9 aut.</sZ>
<sZ>10 aut.</sZ>
<sZ>14 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Mt. Sinai Hospital</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Tellier, R" sort="Tellier, R" uniqKey="Tellier R" first="R." last="Tellier">R. Tellier</name>
<affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Hospital for Sick Children</s1>
<s3>CAN</s3>
<sZ>5 aut.</sZ>
<sZ>15 aut.</sZ>
<sZ>19 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Hospital for Sick Children</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Smieja, M" sort="Smieja, M" uniqKey="Smieja M" first="M." last="Smieja">M. Smieja</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>St. Joseph's Hospital</s1>
<s2>Hamilton, Ontario</s2>
<s3>CAN</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>8 aut.</sZ>
<sZ>16 aut.</sZ>
<sZ>17 aut.</sZ>
<sZ>18 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>St. Joseph's Hospital</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Cai, W" sort="Cai, W" uniqKey="Cai W" first="W." last="Cai">W. Cai</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>St. Joseph's Hospital</s1>
<s2>Hamilton, Ontario</s2>
<s3>CAN</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>8 aut.</sZ>
<sZ>16 aut.</sZ>
<sZ>17 aut.</sZ>
<sZ>18 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>St. Joseph's Hospital</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chernesky, M" sort="Chernesky, M" uniqKey="Chernesky M" first="M." last="Chernesky">M. Chernesky</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>St. Joseph's Hospital</s1>
<s2>Hamilton, Ontario</s2>
<s3>CAN</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>8 aut.</sZ>
<sZ>16 aut.</sZ>
<sZ>17 aut.</sZ>
<sZ>18 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>St. Joseph's Hospital</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Richardson, S E" sort="Richardson, S E" uniqKey="Richardson S" first="S. E." last="Richardson">S. E. Richardson</name>
<affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Hospital for Sick Children</s1>
<s3>CAN</s3>
<sZ>5 aut.</sZ>
<sZ>15 aut.</sZ>
<sZ>19 aut.</sZ>
</inist:fA14>
<country>Canada</country>
<wicri:noRegion>Hospital for Sick Children</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j" type="main">Journal of clinical microbiology : (Print)</title>
<title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title>
<idno type="ISSN">0095-1137</idno>
<imprint><date when="2006">2006</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><title level="j" type="main">Journal of clinical microbiology : (Print)</title>
<title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title>
<idno type="ISSN">0095-1137</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus</term>
<term>Detection</term>
<term>Feces</term>
<term>Feces (virology)</term>
<term>Humans</term>
<term>Method</term>
<term>Microbiology</term>
<term>Molecular Diagnostic Techniques</term>
<term>Nucleic acid</term>
<term>RNA, Viral (analysis)</term>
<term>RNA, Viral (genetics)</term>
<term>RNA, Viral (isolation & purification)</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (isolation & purification)</term>
<term>Sensitivity and Specificity</term>
<term>Severe Acute Respiratory Syndrome (diagnosis)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Severe acute respiratory syndrome</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (analyse)</term>
<term>ARN viral (génétique)</term>
<term>ARN viral (isolement et purification)</term>
<term>Fèces (virologie)</term>
<term>Humains</term>
<term>RT-PCR</term>
<term>Sensibilité et spécificité</term>
<term>Syndrome respiratoire aigu sévère (diagnostic)</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Techniques de diagnostic moléculaire</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus du SRAS (isolement et purification)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN viral</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>ARN viral</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Fèces</term>
<term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Feces</term>
<term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Humans</term>
<term>Molecular Diagnostic Techniques</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>Sensitivity and Specificity</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term>
<term>Acide nucléique</term>
<term>Humains</term>
<term>Méthode</term>
<term>Détection</term>
<term>Fèces</term>
<term>Microbiologie</term>
<term>RT-PCR</term>
<term>Sensibilité et spécificité</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Techniques de diagnostic moléculaire</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">The emergence of a novel coronavirus (CoV) as the cause of severe acute respiratory syndrome (SARS) catalyzed the development of rapid diagnostic tests. Stool samples have been shown to be appropriate for diagnostic testing for SARS CoV, although it has been recognized to be a heterogeneous and difficult sample that contains amplification inhibitors. Limited information on the efficiency of extraction methods for the purification and concentration of SARS CoV RNA from stool samples is available. Our study objectives were to determine the optimal extraction method for SARS CoV RNA detection and to examine the effect of increased specimen volume for the detection of SARS CoV RNA in stool specimens. We conducted a multicenter evaluation of four automated and four manual extraction methods using dilutions of viral lysate in replicate mock stool samples, followed by quantitation of SARS CoV RNA using real-time reverse transcriptase PCR. The sensitivities of the manual methods ranged from 50% to 100%, with the Cortex Biochem Magazorb method, a magnetic bead isolation method, allowing detection of all 12 positive samples. The sensitivities of the automated methods ranged from 75% to 100%. The bioMérieux NucliSens automated extractor and miniMag extraction methods each had a sensitivity of 100%. Examination of the copy numbers detected and the generation of 10-fold dilutions of the extracted material indicated that a number of extraction methods retained inhibitory substances that prevented optimal amplification. Increasing the volume of sample input did improve detection. This information could be useful for the extraction of other RNA viruses from stool samples and demonstrates the need to evaluate extraction methods for different specimen types.</div>
</front>
</TEI>
<affiliations><list><country><li>Canada</li>
</country>
<region><li>Ontario</li>
</region>
<settlement><li>Hamilton (Ontario)</li>
<li>Toronto</li>
</settlement>
<orgName><li>Université McMaster</li>
<li>Université de Toronto</li>
</orgName>
</list>
<tree><country name="Canada"><noRegion><name sortKey="Petrich, A" sort="Petrich, A" uniqKey="Petrich A" first="A." last="Petrich">A. Petrich</name>
</noRegion>
<name sortKey="Akhaven, P" sort="Akhaven, P" uniqKey="Akhaven P" first="P." last="Akhaven">P. Akhaven</name>
<name sortKey="Broukhanski, G" sort="Broukhanski, G" uniqKey="Broukhanski G" first="G." last="Broukhanski">G. Broukhanski</name>
<name sortKey="Cai, W" sort="Cai, W" uniqKey="Cai W" first="W." last="Cai">W. Cai</name>
<name sortKey="Chernesky, M" sort="Chernesky, M" uniqKey="Chernesky M" first="M." last="Chernesky">M. Chernesky</name>
<name sortKey="Chong, S" sort="Chong, S" uniqKey="Chong S" first="S." last="Chong">S. Chong</name>
<name sortKey="Chui, L" sort="Chui, L" uniqKey="Chui L" first="L." last="Chui">L. Chui</name>
<name sortKey="Chui, L" sort="Chui, L" uniqKey="Chui L" first="L." last="Chui">L. Chui</name>
<name sortKey="Chui, L" sort="Chui, L" uniqKey="Chui L" first="L." last="Chui">L. Chui</name>
<name sortKey="Gharabaghi, F" sort="Gharabaghi, F" uniqKey="Gharabaghi F" first="F." last="Gharabaghi">F. Gharabaghi</name>
<name sortKey="Jamieson, F" sort="Jamieson, F" uniqKey="Jamieson F" first="F." last="Jamieson">F. Jamieson</name>
<name sortKey="Johnson, G" sort="Johnson, G" uniqKey="Johnson G" first="G." last="Johnson">G. Johnson</name>
<name sortKey="Louie, L" sort="Louie, L" uniqKey="Louie L" first="L." last="Louie">L. Louie</name>
<name sortKey="Louie, M" sort="Louie, M" uniqKey="Louie M" first="M." last="Louie">M. Louie</name>
<name sortKey="Luinstra, K" sort="Luinstra, K" uniqKey="Luinstra K" first="K." last="Luinstra">K. Luinstra</name>
<name sortKey="Mahony, J" sort="Mahony, J" uniqKey="Mahony J" first="J." last="Mahony">J. Mahony</name>
<name sortKey="Mazzulli, T" sort="Mazzulli, T" uniqKey="Mazzulli T" first="T." last="Mazzulli">T. Mazzulli</name>
<name sortKey="Richardson, S E" sort="Richardson, S E" uniqKey="Richardson S" first="S. E." last="Richardson">S. E. Richardson</name>
<name sortKey="Smieja, M" sort="Smieja, M" uniqKey="Smieja M" first="M." last="Smieja">M. Smieja</name>
<name sortKey="Tellier, R" sort="Tellier, R" uniqKey="Tellier R" first="R." last="Tellier">R. Tellier</name>
<name sortKey="Willey, B" sort="Willey, B" uniqKey="Willey B" first="B." last="Willey">B. Willey</name>
</country>
</tree>
</affiliations>
</record>
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