SrasV1, PascalFrancis, Corpus, bibRecord, 000469

Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens

Identifieur interne : 000469 ( PascalFrancis/Corpus ); précédent : 000468; suivant : 000470

Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens

Auteurs : A. Petrich ; J. Mahony ; S. Chong ; G. Broukhanski ; F. Gharabaghi ; G. Johnson ; L. Louie ; K. Luinstra ; B. Willey ; P. Akhaven ; L. Chui ; F. Jamieson ; M. Louie ; T. Mazzulli ; R. Tellier ; M. Smieja ; W. Cai ; M. Chernesky ; S. E. Richardson

Source :

Journal of clinical microbiology : (Print) [ 0095-1137 ] ; 2006.

RBID : Pascal:06-0404132

Descripteurs français

Pascal (Inist)
- Coronavirus, Acide nucléique, Méthode, Détection, Fèces, Microbiologie, Syndrome respiratoire aigu sévère.

English descriptors

KwdEn :
- Coronavirus, Detection, Feces, Method, Microbiology, Nucleic acid, Severe acute respiratory syndrome.

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.

Notice en format standard (ISO 2709)

Pour connaître la documentation sur le format Inist Standard.

A01	`01`	`1`		`@0 0095-1137`
A02	`01`			`@0 JCMIDW`
A03		`1`		`@0 J. clin. microbiol. : (Print)`
A05				`@2 44`
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A08	`01`	`1`	`ENG`	`@1 Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens`
A11	`01`	`1`		`@1 PETRICH (A.)`
A11	`02`	`1`		`@1 MAHONY (J.)`
A11	`03`	`1`		`@1 CHONG (S.)`
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A11	`05`	`1`		`@1 GHARABAGHI (F.)`
A11	`06`	`1`		`@1 JOHNSON (G.)`
A11	`07`	`1`		`@1 LOUIE (L.)`
A11	`08`	`1`		`@1 LUINSTRA (K.)`
A11	`09`	`1`		`@1 WILLEY (B.)`
A11	`10`	`1`		`@1 AKHAVEN (P.)`
A11	`11`	`1`		`@1 CHUI (L.)`
A11	`12`	`1`		`@1 JAMIESON (F.)`
A11	`13`	`1`		`@1 LOUIE (M.)`
A11	`14`	`1`		`@1 MAZZULLI (T.)`
A11	`15`	`1`		`@1 TELLIER (R.)`
A11	`16`	`1`		`@1 SMIEJA (M.)`
A11	`17`	`1`		`@1 CAI (W.)`
A11	`18`	`1`		`@1 CHERNESKY (M.)`
A11	`19`	`1`		`@1 RICHARDSON (S. E.)`
A14	`01`			`@1 St. Joseph's Hospital @2 Hamilton, Ontario @3 CAN @Z 1 aut. @Z 2 aut. @Z 3 aut. @Z 8 aut. @Z 16 aut. @Z 17 aut. @Z 18 aut.`
A14	`02`			`@1 Ontario Ministry of Health @2 Etobicoke, Ontario @3 CAN @Z 4 aut. @Z 12 aut.`
A14	`03`			`@1 Hospital for Sick Children @3 CAN @Z 5 aut. @Z 15 aut. @Z 19 aut.`
A14	`04`			`@1 St. Michael's Hospital @3 CAN @Z 6 aut.`
A14	`05`			`@1 Sunny Brook & WHC HSC @3 CAN @Z 7 aut.`
A14	`06`			`@1 Mt. Sinai Hospital @2 Toronto, Ontario @3 CAN @Z 9 aut. @Z 10 aut. @Z 14 aut.`
A14	`07`			`@1 Edmonton Public Health Laboratory @2 Edmonton, Alberta @3 CAN @Z 11 aut.`
A14	`08`			`@1 Calgary Public Health Laboratory @2 Calgary, Alberta @3 CAN @Z 11 aut.`
A14	`09`			`@1 McMaster University @2 Hamilton, Ontario @3 CAN @Z 11 aut.`
A14	`10`			`@1 The University of Toronto @2 Toronto, Ontario @3 CAN @Z 13 aut.`
A17	`01`	`1`		`@1 Ontario Laboratory Working Group for the Rapid Diagnosis of Emerging Infections @3 CAN`
A20				`@1 2681-2688`
A21				`@1 2006`
A23	`01`			`@0 ENG`
A43	`01`			`@1 INIST @2 17088 @5 354000133476880020`
A44				`@0 0000 @1 © 2006 INIST-CNRS. All rights reserved.`
A45				`@0 12 ref.`
A47	`01`	`1`		`@0 06-0404132`
A60				`@1 P`
A61				`@0 A`
A64	`01`	`1`		`@0 Journal of clinical microbiology : (Print)`
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C01	`01`		`ENG`	@0 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.
C02	`01`	`X`		`@0 002A05C10`
C02	`02`	`X`		`@0 002B05`
C03	`01`	`X`	`FRE`	`@0 Coronavirus @2 NW @5 01`
C03	`01`	`X`	`ENG`	`@0 Coronavirus @2 NW @5 01`
C03	`01`	`X`	`SPA`	`@0 Coronavirus @2 NW @5 01`
C03	`02`	`X`	`FRE`	`@0 Acide nucléique @5 05`
C03	`02`	`X`	`ENG`	`@0 Nucleic acid @5 05`
C03	`02`	`X`	`SPA`	`@0 Acido nucleico @5 05`
C03	`03`	`X`	`FRE`	`@0 Méthode @5 06`
C03	`03`	`X`	`ENG`	`@0 Method @5 06`
C03	`03`	`X`	`SPA`	`@0 Método @5 06`
C03	`04`	`X`	`FRE`	`@0 Détection @5 07`
C03	`04`	`X`	`ENG`	`@0 Detection @5 07`
C03	`04`	`X`	`SPA`	`@0 Detección @5 07`
C03	`05`	`X`	`FRE`	`@0 Fèces @5 08`
C03	`05`	`X`	`ENG`	`@0 Feces @5 08`
C03	`05`	`X`	`SPA`	`@0 Heces @5 08`
C03	`06`	`X`	`FRE`	`@0 Microbiologie @5 09`
C03	`06`	`X`	`ENG`	`@0 Microbiology @5 09`
C03	`06`	`X`	`SPA`	`@0 Microbiología @5 09`
C03	`07`	`X`	`FRE`	`@0 Syndrome respiratoire aigu sévère @2 NM @5 14`
C03	`07`	`X`	`ENG`	`@0 Severe acute respiratory syndrome @2 NM @5 14`
C03	`07`	`X`	`SPA`	`@0 Síndrome respiratorio agudo severo @2 NM @5 14`
C07	`01`	`X`	`FRE`	`@0 Coronaviridae @2 NW`
C07	`01`	`X`	`ENG`	`@0 Coronaviridae @2 NW`
C07	`01`	`X`	`SPA`	`@0 Coronaviridae @2 NW`
C07	`02`	`X`	`FRE`	`@0 Nidovirales @2 NW`
C07	`02`	`X`	`ENG`	`@0 Nidovirales @2 NW`
C07	`02`	`X`	`SPA`	`@0 Nidovirales @2 NW`
C07	`03`	`X`	`FRE`	`@0 Virus @2 NW`
C07	`03`	`X`	`ENG`	`@0 Virus @2 NW`
C07	`03`	`X`	`SPA`	`@0 Virus @2 NW`
C07	`04`	`X`	`FRE`	`@0 Appareil respiratoire pathologie @5 13`
C07	`04`	`X`	`ENG`	`@0 Respiratory disease @5 13`
C07	`04`	`X`	`SPA`	`@0 Aparato respiratorio patología @5 13`
C07	`05`	`X`	`FRE`	`@0 Virose`
C07	`05`	`X`	`ENG`	`@0 Viral disease`
C07	`05`	`X`	`SPA`	`@0 Virosis`
C07	`06`	`X`	`FRE`	`@0 Infection`
C07	`06`	`X`	`ENG`	`@0 Infection`
C07	`06`	`X`	`SPA`	`@0 Infección`
C07	`07`	`X`	`FRE`	`@0 Poumon pathologie @5 16`
C07	`07`	`X`	`ENG`	`@0 Lung disease @5 16`
C07	`07`	`X`	`SPA`	`@0 Pulmón patología @5 16`
N21				`@1 268`
N44	`01`			`@1 OTO`
N82				`@1 OTO`

Format Inist (serveur)

NO :	PASCAL 06-0404132 INIST
ET :	Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens
AU :	PETRICH (A.); MAHONY (J.); CHONG (S.); BROUKHANSKI (G.); GHARABAGHI (F.); JOHNSON (G.); LOUIE (L.); LUINSTRA (K.); WILLEY (B.); AKHAVEN (P.); CHUI (L.); JAMIESON (F.); LOUIE (M.); MAZZULLI (T.); TELLIER (R.); SMIEJA (M.); CAI (W.); CHERNESKY (M.); RICHARDSON (S. E.)
AF :	St. Joseph's Hospital/Hamilton, Ontario/Canada (1 aut., 2 aut., 3 aut., 8 aut., 16 aut., 17 aut., 18 aut.); Ontario Ministry of Health/Etobicoke, Ontario/Canada (4 aut., 12 aut.); Hospital for Sick Children/Canada (5 aut., 15 aut., 19 aut.); St. Michael's Hospital/Canada (6 aut.); Sunny Brook & WHC HSC/Canada (7 aut.); Mt. Sinai Hospital/Toronto, Ontario/Canada (9 aut., 10 aut., 14 aut.); Edmonton Public Health Laboratory/Edmonton, Alberta/Canada (11 aut.); Calgary Public Health Laboratory/Calgary, Alberta/Canada (11 aut.); McMaster University/Hamilton, Ontario/Canada (11 aut.); The University of Toronto/Toronto, Ontario/Canada (13 aut.)
DT :	Publication en série; Niveau analytique
SO :	Journal of clinical microbiology : (Print); ISSN 0095-1137; Coden JCMIDW; Etats-Unis; Da. 2006; Vol. 44; No. 8; Pp. 2681-2688; Bibl. 12 ref.
LA :	Anglais
EA :	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.
CC :	002A05C10; 002B05
FD :	Coronavirus; Acide nucléique; Méthode; Détection; Fèces; Microbiologie; Syndrome respiratoire aigu sévère
FG :	Coronaviridae; Nidovirales; Virus; Appareil respiratoire pathologie; Virose; Infection; Poumon pathologie
ED :	Coronavirus; Nucleic acid; Method; Detection; Feces; Microbiology; Severe acute respiratory syndrome
EG :	Coronaviridae; Nidovirales; Virus; Respiratory disease; Viral disease; Infection; Lung disease
SD :	Coronavirus; Acido nucleico; Método; Detección; Heces; Microbiología; Síndrome respiratorio agudo severo
LO :	INIST-17088.354000133476880020
ID :	06-0404132

Links to Exploration step

Pascal:06-0404132

Le document en format XML

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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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<author><name sortKey="Cai, W" sort="Cai, W" uniqKey="Cai W" first="W." last="Cai">W. Cai</name>
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<author><name sortKey="Chernesky, M" sort="Chernesky, M" uniqKey="Chernesky M" first="M." last="Chernesky">M. Chernesky</name>
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<author><name sortKey="Richardson, S E" sort="Richardson, S E" uniqKey="Richardson S" first="S. E." last="Richardson">S. E. Richardson</name>
<affiliation><inist:fA14 i1="03"><s1>Hospital for Sick Children</s1>
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<idno type="inist">06-0404132</idno>
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<idno type="RBID">Pascal:06-0404132</idno>
<idno type="wicri:Area/PascalFrancis/Corpus">000469</idno>
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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>
<author><name sortKey="Petrich, A" sort="Petrich, A" uniqKey="Petrich A" first="A." last="Petrich">A. Petrich</name>
<affiliation><inist:fA14 i1="01"><s1>St. Joseph's Hospital</s1>
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<s3>CAN</s3>
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<s3>CAN</s3>
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<s3>CAN</s3>
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<s3>CAN</s3>
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<s3>CAN</s3>
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<s3>CAN</s3>
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<s3>CAN</s3>
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<affiliation><inist:fA14 i1="10"><s1>The University of Toronto</s1>
<s2>Toronto, Ontario</s2>
<s3>CAN</s3>
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<author><name sortKey="Mazzulli, T" sort="Mazzulli, T" uniqKey="Mazzulli T" first="T." last="Mazzulli">T. Mazzulli</name>
<affiliation><inist:fA14 i1="06"><s1>Mt. Sinai Hospital</s1>
<s2>Toronto, Ontario</s2>
<s3>CAN</s3>
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<sZ>10 aut.</sZ>
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<author><name sortKey="Tellier, R" sort="Tellier, R" uniqKey="Tellier R" first="R." last="Tellier">R. Tellier</name>
<affiliation><inist:fA14 i1="03"><s1>Hospital for Sick Children</s1>
<s3>CAN</s3>
<sZ>5 aut.</sZ>
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<affiliation><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>
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<author><name sortKey="Cai, W" sort="Cai, W" uniqKey="Cai W" first="W." last="Cai">W. Cai</name>
<affiliation><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>
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<author><name sortKey="Chernesky, M" sort="Chernesky, M" uniqKey="Chernesky M" first="M." last="Chernesky">M. Chernesky</name>
<affiliation><inist:fA14 i1="01"><s1>St. Joseph's Hospital</s1>
<s2>Hamilton, Ontario</s2>
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<sZ>1 aut.</sZ>
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<author><name sortKey="Richardson, S E" sort="Richardson, S E" uniqKey="Richardson S" first="S. E." last="Richardson">S. E. Richardson</name>
<affiliation><inist:fA14 i1="03"><s1>Hospital for Sick Children</s1>
<s3>CAN</s3>
<sZ>5 aut.</sZ>
<sZ>15 aut.</sZ>
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<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>
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<seriesStmt><title level="j" type="main">Journal of clinical microbiology : (Print)</title>
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<idno type="ISSN">0095-1137</idno>
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<term>Detection</term>
<term>Feces</term>
<term>Method</term>
<term>Microbiology</term>
<term>Nucleic acid</term>
<term>Severe acute respiratory syndrome</term>
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<keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term>
<term>Acide nucléique</term>
<term>Méthode</term>
<term>Détection</term>
<term>Fèces</term>
<term>Microbiologie</term>
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<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>
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<fA08 i1="01" i2="1" l="ENG"><s1>Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens</s1>
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<fA11 i1="01" i2="1"><s1>PETRICH (A.)</s1>
</fA11>
<fA11 i1="02" i2="1"><s1>MAHONY (J.)</s1>
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<fA11 i1="03" i2="1"><s1>CHONG (S.)</s1>
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<fA11 i1="04" i2="1"><s1>BROUKHANSKI (G.)</s1>
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<fA11 i1="13" i2="1"><s1>LOUIE (M.)</s1>
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<fA11 i1="14" i2="1"><s1>MAZZULLI (T.)</s1>
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<fA11 i1="16" i2="1"><s1>SMIEJA (M.)</s1>
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<sZ>5 aut.</sZ>
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<sZ>7 aut.</sZ>
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<s3>CAN</s3>
<sZ>9 aut.</sZ>
<sZ>10 aut.</sZ>
<sZ>14 aut.</sZ>
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<fA14 i1="07"><s1>Edmonton Public Health Laboratory</s1>
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<s3>CAN</s3>
<sZ>13 aut.</sZ>
</fA14>
<fA17 i1="01" i2="1"><s1>Ontario Laboratory Working Group for the Rapid Diagnosis of Emerging Infections</s1>
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<fC01 i1="01" l="ENG"><s0>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.</s0>
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<fC07 i1="06" i2="X" l="SPA"><s0>Infección</s0>
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<s5>16</s5>
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<s5>16</s5>
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<s5>16</s5>
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<server><NO>PASCAL 06-0404132 INIST</NO>
<ET>Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens</ET>
<AU>PETRICH (A.); MAHONY (J.); CHONG (S.); BROUKHANSKI (G.); GHARABAGHI (F.); JOHNSON (G.); LOUIE (L.); LUINSTRA (K.); WILLEY (B.); AKHAVEN (P.); CHUI (L.); JAMIESON (F.); LOUIE (M.); MAZZULLI (T.); TELLIER (R.); SMIEJA (M.); CAI (W.); CHERNESKY (M.); RICHARDSON (S. E.)</AU>
<AF>St. Joseph's Hospital/Hamilton, Ontario/Canada (1 aut., 2 aut., 3 aut., 8 aut., 16 aut., 17 aut., 18 aut.); Ontario Ministry of Health/Etobicoke, Ontario/Canada (4 aut., 12 aut.); Hospital for Sick Children/Canada (5 aut., 15 aut., 19 aut.); St. Michael's Hospital/Canada (6 aut.); Sunny Brook & WHC HSC/Canada (7 aut.); Mt. Sinai Hospital/Toronto, Ontario/Canada (9 aut., 10 aut., 14 aut.); Edmonton Public Health Laboratory/Edmonton, Alberta/Canada (11 aut.); Calgary Public Health Laboratory/Calgary, Alberta/Canada (11 aut.); McMaster University/Hamilton, Ontario/Canada (11 aut.); The University of Toronto/Toronto, Ontario/Canada (13 aut.)</AF>
<DT>Publication en série; Niveau analytique</DT>
<SO>Journal of clinical microbiology : (Print); ISSN 0095-1137; Coden JCMIDW; Etats-Unis; Da. 2006; Vol. 44; No. 8; Pp. 2681-2688; Bibl. 12 ref.</SO>
<LA>Anglais</LA>
<EA>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.</EA>
<CC>002A05C10; 002B05</CC>
<FD>Coronavirus; Acide nucléique; Méthode; Détection; Fèces; Microbiologie; Syndrome respiratoire aigu sévère</FD>
<FG>Coronaviridae; Nidovirales; Virus; Appareil respiratoire pathologie; Virose; Infection; Poumon pathologie</FG>
<ED>Coronavirus; Nucleic acid; Method; Detection; Feces; Microbiology; Severe acute respiratory syndrome</ED>
<EG>Coronaviridae; Nidovirales; Virus; Respiratory disease; Viral disease; Infection; Lung disease</EG>
<SD>Coronavirus; Acido nucleico; Método; Detección; Heces; Microbiología; Síndrome respiratorio agudo severo</SD>
<LO>INIST-17088.354000133476880020</LO>
<ID>06-0404132</ID>
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Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens

Multicenter comparison of nucleic acid extraction methods for detection of severe acute respiratory syndrome coronavirus RNA in stool specimens

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