PandemieGrippaleV1, PascalFrancis, Curation, bibRecord, 000C43

Development of a multiplex real-time RT-PCR assay for detection of influenza A, influenza B, RSV and typing of the 2009-H1N1 influenza virus

Identifieur interne : 000C43 ( PascalFrancis/Curation ); précédent : 000C42; suivant : 000C44

Development of a multiplex real-time RT-PCR assay for detection of influenza A, influenza B, RSV and typing of the 2009-H1N1 influenza virus

Auteurs : Weston C. Hymas [États-Unis] ; Alan Mills [États-Unis] ; Sheri Ferguson [États-Unis] ; Janine Langer [États-Unis] ; Rosemary C. She [États-Unis] ; Walt Mahoney [États-Unis] ; David R. Hillyard [États-Unis]

Source :

Journal of virological methods [ 0166-0934 ] ; 2010.

RBID : Pascal:10-0326823

Descripteurs français

Pascal (Inist)
- Virus grippal A, Réaction chaîne polymérase multiplex, Temps réel, Réaction chaîne polymérase RT, Détection, Typage, Polymorphisme mononucléotide, Méthode, Grippe A, Grippe B, Virus grippal A(H1N1).

English descriptors

KwdEn :
- Detection, Influenza A, Influenza A virus, Influenza B, Influenzavirus A(H1N1), Method, Multiplex polymerase chain reaction, Real time, Reverse transcription polymerase chain reaction, Single nucleotide polymorphism, Typing.

Abstract

A high-throughput real-time RT-PCR assay was developed to amplify and detect a conserved region of the hemagglutinin gene of the 2009-H1N1 influenza A virus using a minor groove binder-conjugated hybridization probe. The assay was paired with a separate triplex real-time assay that detects influenza A via the matrix gene, influenza B and RSV in a multiplex format and compared with the Centers for Disease Control and Prevention (CDC) rRT-PCR assay using 143 samples. The 2009-H1N1 portion of the multiplex assay had 100% correlation with the CDC assay, while the triplex assay had a 99% agreement. An additional 105 samples collected from October to November 2009 were also tested using both the individual 2009-H1N1 and triplex assays. Of these 105 samples, eight were positive for the hemagglutinin target in the H1N1 assay and negative for the matrix target in the triplex assay. Discrepant analysis revealed single nucleotide polymorphisms within the matrix gene of 2009-H1N1 virus-positive samples. The limit of detection for the 2009-H1N1 assay was between 750 and 1500 copies/reaction and no cross-reactivity with other respiratory pathogens was observed. Overall, this multiplexed format proved to be sensitive, robust and easy to use and serves as a useful tool for pandemic testing.

A01	`01`	`1`		`@0 0166-0934`
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A03		`1`		`@0 J. virol. methods`
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A08	`01`	`1`	`ENG`	`@1 Development of a multiplex real-time RT-PCR assay for detection of influenza A, influenza B, RSV and typing of the 2009-H1N1 influenza virus`
A11	`01`	`1`		`@1 HYMAS (Weston C.)`
A11	`02`	`1`		`@1 MILLS (Alan)`
A11	`03`	`1`		`@1 FERGUSON (Sheri)`
A11	`04`	`1`		`@1 LANGER (Janine)`
A11	`05`	`1`		`@1 SHE (Rosemary C.)`
A11	`06`	`1`		`@1 MAHONEY (Walt)`
A11	`07`	`1`		`@1 HILLYARD (David R.)`
A14	`01`			`@1 Associated Regional and University Pathologists (ARUP) Institute for Clinical and Experimental Pathology, 500 Chipeta Way @2 Salt Lake City, UT 84108 @3 USA @Z 1 aut. @Z 5 aut. @Z 7 aut.`
A14	`02`			`@1 Epoch Biosciences, A Division of Wescor, 21720 23rd Drive S.E. Suite 150 @2 Bothell, WA 98021 @3 USA @Z 2 aut. @Z 6 aut.`
A14	`03`			`@1 Associated Regional and University Pathologists (ARUP), 500 Chipeta Way @2 Salt Lake City, UT 84108 @3 USA @Z 3 aut. @Z 4 aut.`
A14	`04`			`@1 Department of Pathology, University of Utah Health Sciences Center, 15N. Medical Drive East, 1100EEJ @2 Salt Lake City, UT 84112 @3 USA @Z 5 aut. @Z 7 aut.`
A20				`@1 113-118`
A21				`@1 2010`
A23	`01`			`@0 ENG`
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A44				`@0 0000 @1 © 2010 INIST-CNRS. All rights reserved.`
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A64	`01`	`1`		`@0 Journal of virological methods`
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C01	`01`		`ENG`	@0 A high-throughput real-time RT-PCR assay was developed to amplify and detect a conserved region of the hemagglutinin gene of the 2009-H1N1 influenza A virus using a minor groove binder-conjugated hybridization probe. The assay was paired with a separate triplex real-time assay that detects influenza A via the matrix gene, influenza B and RSV in a multiplex format and compared with the Centers for Disease Control and Prevention (CDC) rRT-PCR assay using 143 samples. The 2009-H1N1 portion of the multiplex assay had 100% correlation with the CDC assay, while the triplex assay had a 99% agreement. An additional 105 samples collected from October to November 2009 were also tested using both the individual 2009-H1N1 and triplex assays. Of these 105 samples, eight were positive for the hemagglutinin target in the H1N1 assay and negative for the matrix target in the triplex assay. Discrepant analysis revealed single nucleotide polymorphisms within the matrix gene of 2009-H1N1 virus-positive samples. The limit of detection for the 2009-H1N1 assay was between 750 and 1500 copies/reaction and no cross-reactivity with other respiratory pathogens was observed. Overall, this multiplexed format proved to be sensitive, robust and easy to use and serves as a useful tool for pandemic testing.
C02	`01`	`X`		`@0 002A05C09`
C03	`01`	`X`	`FRE`	`@0 Virus grippal A @2 NW @5 01`
C03	`01`	`X`	`ENG`	`@0 Influenza A virus @2 NW @5 01`
C03	`01`	`X`	`SPA`	`@0 Influenza A virus @2 NW @5 01`
C03	`02`	`X`	`FRE`	`@0 Réaction chaîne polymérase multiplex @5 05`
C03	`02`	`X`	`ENG`	`@0 Multiplex polymerase chain reaction @5 05`
C03	`02`	`X`	`SPA`	`@0 Reacción cadena polimerasa multiplex @5 05`
C03	`03`	`X`	`FRE`	`@0 Temps réel @5 06`
C03	`03`	`X`	`ENG`	`@0 Real time @5 06`
C03	`03`	`X`	`SPA`	`@0 Tiempo real @5 06`
C03	`04`	`X`	`FRE`	`@0 Réaction chaîne polymérase RT @5 07`
C03	`04`	`X`	`ENG`	`@0 Reverse transcription polymerase chain reaction @5 07`
C03	`04`	`X`	`SPA`	`@0 Reacción cadena polimerasa transcripción inversa @5 07`
C03	`05`	`X`	`FRE`	`@0 Détection @5 08`
C03	`05`	`X`	`ENG`	`@0 Detection @5 08`
C03	`05`	`X`	`SPA`	`@0 Detección @5 08`
C03	`06`	`X`	`FRE`	`@0 Typage @5 09`
C03	`06`	`X`	`ENG`	`@0 Typing @5 09`
C03	`06`	`X`	`SPA`	`@0 Tipificación @5 09`
C03	`07`	`X`	`FRE`	`@0 Polymorphisme mononucléotide @5 10`
C03	`07`	`X`	`ENG`	`@0 Single nucleotide polymorphism @5 10`
C03	`07`	`X`	`SPA`	`@0 Polimorfismo mononucleótido @5 10`
C03	`08`	`X`	`FRE`	`@0 Méthode @5 11`
C03	`08`	`X`	`ENG`	`@0 Method @5 11`
C03	`08`	`X`	`SPA`	`@0 Método @5 11`
C03	`09`	`X`	`FRE`	`@0 Grippe A @5 14`
C03	`09`	`X`	`ENG`	`@0 Influenza A @5 14`
C03	`09`	`X`	`SPA`	`@0 Gripe A @5 14`
C03	`10`	`X`	`FRE`	`@0 Grippe B @5 15`
C03	`10`	`X`	`ENG`	`@0 Influenza B @5 15`
C03	`10`	`X`	`SPA`	`@0 Gripe B @5 15`
C03	`11`	`X`	`FRE`	`@0 Virus grippal A(H1N1) @4 CD @5 96`
C03	`11`	`X`	`ENG`	`@0 Influenzavirus A(H1N1) @4 CD @5 96`
C07	`01`	`X`	`FRE`	`@0 Influenzavirus A @2 NW`
C07	`01`	`X`	`ENG`	`@0 Influenzavirus A @2 NW`
C07	`01`	`X`	`SPA`	`@0 Influenzavirus A @2 NW`
C07	`02`	`X`	`FRE`	`@0 Orthomyxoviridae @2 NW`
C07	`02`	`X`	`ENG`	`@0 Orthomyxoviridae @2 NW`
C07	`02`	`X`	`SPA`	`@0 Orthomyxoviridae @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 Virose`
C07	`04`	`X`	`ENG`	`@0 Viral disease`
C07	`04`	`X`	`SPA`	`@0 Virosis`
C07	`05`	`X`	`FRE`	`@0 Infection`
C07	`05`	`X`	`ENG`	`@0 Infection`
C07	`05`	`X`	`SPA`	`@0 Infección`
N21				`@1 207`
N44	`01`			`@1 OTO`
N82				`@1 OTO`

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Pascal:10-0326823

Le document en format XML

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<series><title level="j" type="main">Journal of virological methods</title>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Detection</term>
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<term>Influenzavirus A(H1N1)</term>
<term>Method</term>
<term>Multiplex polymerase chain reaction</term>
<term>Real time</term>
<term>Reverse transcription polymerase chain reaction</term>
<term>Single nucleotide polymorphism</term>
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<keywords scheme="Pascal" xml:lang="fr"><term>Virus grippal A</term>
<term>Réaction chaîne polymérase multiplex</term>
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<term>Réaction chaîne polymérase RT</term>
<term>Détection</term>
<term>Typage</term>
<term>Polymorphisme mononucléotide</term>
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<front><div type="abstract" xml:lang="en">A high-throughput real-time RT-PCR assay was developed to amplify and detect a conserved region of the hemagglutinin gene of the 2009-H1N1 influenza A virus using a minor groove binder-conjugated hybridization probe. The assay was paired with a separate triplex real-time assay that detects influenza A via the matrix gene, influenza B and RSV in a multiplex format and compared with the Centers for Disease Control and Prevention (CDC) rRT-PCR assay using 143 samples. The 2009-H1N1 portion of the multiplex assay had 100% correlation with the CDC assay, while the triplex assay had a 99% agreement. An additional 105 samples collected from October to November 2009 were also tested using both the individual 2009-H1N1 and triplex assays. Of these 105 samples, eight were positive for the hemagglutinin target in the H1N1 assay and negative for the matrix target in the triplex assay. Discrepant analysis revealed single nucleotide polymorphisms within the matrix gene of 2009-H1N1 virus-positive samples. The limit of detection for the 2009-H1N1 assay was between 750 and 1500 copies/reaction and no cross-reactivity with other respiratory pathogens was observed. Overall, this multiplexed format proved to be sensitive, robust and easy to use and serves as a useful tool for pandemic testing.</div>
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<sZ>7 aut.</sZ>
</fA14>
<fA14 i1="02"><s1>Epoch Biosciences, A Division of Wescor, 21720 23rd Drive S.E. Suite 150</s1>
<s2>Bothell, WA 98021</s2>
<s3>USA</s3>
<sZ>2 aut.</sZ>
<sZ>6 aut.</sZ>
</fA14>
<fA14 i1="03"><s1>Associated Regional and University Pathologists (ARUP), 500 Chipeta Way</s1>
<s2>Salt Lake City, UT 84108</s2>
<s3>USA</s3>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
</fA14>
<fA14 i1="04"><s1>Department of Pathology, University of Utah Health Sciences Center, 15N. Medical Drive East, 1100EEJ</s1>
<s2>Salt Lake City, UT 84112</s2>
<s3>USA</s3>
<sZ>5 aut.</sZ>
<sZ>7 aut.</sZ>
</fA14>
<fA20><s1>113-118</s1>
</fA20>
<fA21><s1>2010</s1>
</fA21>
<fA23 i1="01"><s0>ENG</s0>
</fA23>
<fA43 i1="01"><s1>INIST</s1>
<s2>18295</s2>
<s5>354000192597880010</s5>
</fA43>
<fA44><s0>0000</s0>
<s1>© 2010 INIST-CNRS. All rights reserved.</s1>
</fA44>
<fA45><s0>3/4 p.</s0>
</fA45>
<fA47 i1="01" i2="1"><s0>10-0326823</s0>
</fA47>
<fA60><s1>P</s1>
<s3>PR</s3>
</fA60>
<fA61><s0>A</s0>
</fA61>
<fA64 i1="01" i2="1"><s0>Journal of virological methods</s0>
</fA64>
<fA66 i1="01"><s0>GBR</s0>
</fA66>
<fC01 i1="01" l="ENG"><s0>A high-throughput real-time RT-PCR assay was developed to amplify and detect a conserved region of the hemagglutinin gene of the 2009-H1N1 influenza A virus using a minor groove binder-conjugated hybridization probe. The assay was paired with a separate triplex real-time assay that detects influenza A via the matrix gene, influenza B and RSV in a multiplex format and compared with the Centers for Disease Control and Prevention (CDC) rRT-PCR assay using 143 samples. The 2009-H1N1 portion of the multiplex assay had 100% correlation with the CDC assay, while the triplex assay had a 99% agreement. An additional 105 samples collected from October to November 2009 were also tested using both the individual 2009-H1N1 and triplex assays. Of these 105 samples, eight were positive for the hemagglutinin target in the H1N1 assay and negative for the matrix target in the triplex assay. Discrepant analysis revealed single nucleotide polymorphisms within the matrix gene of 2009-H1N1 virus-positive samples. The limit of detection for the 2009-H1N1 assay was between 750 and 1500 copies/reaction and no cross-reactivity with other respiratory pathogens was observed. Overall, this multiplexed format proved to be sensitive, robust and easy to use and serves as a useful tool for pandemic testing.</s0>
</fC01>
<fC02 i1="01" i2="X"><s0>002A05C09</s0>
</fC02>
<fC03 i1="01" i2="X" l="FRE"><s0>Virus grippal A</s0>
<s2>NW</s2>
<s5>01</s5>
</fC03>
<fC03 i1="01" i2="X" l="ENG"><s0>Influenza A virus</s0>
<s2>NW</s2>
<s5>01</s5>
</fC03>
<fC03 i1="01" i2="X" l="SPA"><s0>Influenza A virus</s0>
<s2>NW</s2>
<s5>01</s5>
</fC03>
<fC03 i1="02" i2="X" l="FRE"><s0>Réaction chaîne polymérase multiplex</s0>
<s5>05</s5>
</fC03>
<fC03 i1="02" i2="X" l="ENG"><s0>Multiplex polymerase chain reaction</s0>
<s5>05</s5>
</fC03>
<fC03 i1="02" i2="X" l="SPA"><s0>Reacción cadena polimerasa multiplex</s0>
<s5>05</s5>
</fC03>
<fC03 i1="03" i2="X" l="FRE"><s0>Temps réel</s0>
<s5>06</s5>
</fC03>
<fC03 i1="03" i2="X" l="ENG"><s0>Real time</s0>
<s5>06</s5>
</fC03>
<fC03 i1="03" i2="X" l="SPA"><s0>Tiempo real</s0>
<s5>06</s5>
</fC03>
<fC03 i1="04" i2="X" l="FRE"><s0>Réaction chaîne polymérase RT</s0>
<s5>07</s5>
</fC03>
<fC03 i1="04" i2="X" l="ENG"><s0>Reverse transcription polymerase chain reaction</s0>
<s5>07</s5>
</fC03>
<fC03 i1="04" i2="X" l="SPA"><s0>Reacción cadena polimerasa transcripción inversa</s0>
<s5>07</s5>
</fC03>
<fC03 i1="05" i2="X" l="FRE"><s0>Détection</s0>
<s5>08</s5>
</fC03>
<fC03 i1="05" i2="X" l="ENG"><s0>Detection</s0>
<s5>08</s5>
</fC03>
<fC03 i1="05" i2="X" l="SPA"><s0>Detección</s0>
<s5>08</s5>
</fC03>
<fC03 i1="06" i2="X" l="FRE"><s0>Typage</s0>
<s5>09</s5>
</fC03>
<fC03 i1="06" i2="X" l="ENG"><s0>Typing</s0>
<s5>09</s5>
</fC03>
<fC03 i1="06" i2="X" l="SPA"><s0>Tipificación</s0>
<s5>09</s5>
</fC03>
<fC03 i1="07" i2="X" l="FRE"><s0>Polymorphisme mononucléotide</s0>
<s5>10</s5>
</fC03>
<fC03 i1="07" i2="X" l="ENG"><s0>Single nucleotide polymorphism</s0>
<s5>10</s5>
</fC03>
<fC03 i1="07" i2="X" l="SPA"><s0>Polimorfismo mononucleótido</s0>
<s5>10</s5>
</fC03>
<fC03 i1="08" i2="X" l="FRE"><s0>Méthode</s0>
<s5>11</s5>
</fC03>
<fC03 i1="08" i2="X" l="ENG"><s0>Method</s0>
<s5>11</s5>
</fC03>
<fC03 i1="08" i2="X" l="SPA"><s0>Método</s0>
<s5>11</s5>
</fC03>
<fC03 i1="09" i2="X" l="FRE"><s0>Grippe A</s0>
<s5>14</s5>
</fC03>
<fC03 i1="09" i2="X" l="ENG"><s0>Influenza A</s0>
<s5>14</s5>
</fC03>
<fC03 i1="09" i2="X" l="SPA"><s0>Gripe A</s0>
<s5>14</s5>
</fC03>
<fC03 i1="10" i2="X" l="FRE"><s0>Grippe B</s0>
<s5>15</s5>
</fC03>
<fC03 i1="10" i2="X" l="ENG"><s0>Influenza B</s0>
<s5>15</s5>
</fC03>
<fC03 i1="10" i2="X" l="SPA"><s0>Gripe B</s0>
<s5>15</s5>
</fC03>
<fC03 i1="11" i2="X" l="FRE"><s0>Virus grippal A(H1N1)</s0>
<s4>CD</s4>
<s5>96</s5>
</fC03>
<fC03 i1="11" i2="X" l="ENG"><s0>Influenzavirus A(H1N1)</s0>
<s4>CD</s4>
<s5>96</s5>
</fC03>
<fC07 i1="01" i2="X" l="FRE"><s0>Influenzavirus A</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="01" i2="X" l="ENG"><s0>Influenzavirus A</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="01" i2="X" l="SPA"><s0>Influenzavirus A</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="FRE"><s0>Orthomyxoviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="ENG"><s0>Orthomyxoviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="SPA"><s0>Orthomyxoviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="FRE"><s0>Virose</s0>
</fC07>
<fC07 i1="04" i2="X" l="ENG"><s0>Viral disease</s0>
</fC07>
<fC07 i1="04" i2="X" l="SPA"><s0>Virosis</s0>
</fC07>
<fC07 i1="05" i2="X" l="FRE"><s0>Infection</s0>
</fC07>
<fC07 i1="05" i2="X" l="ENG"><s0>Infection</s0>
</fC07>
<fC07 i1="05" i2="X" l="SPA"><s0>Infección</s0>
</fC07>
<fN21><s1>207</s1>
</fN21>
<fN44 i1="01"><s1>OTO</s1>
</fN44>
<fN82><s1>OTO</s1>
</fN82>
</pA>
</standard>
</inist>
</record>

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Development of a multiplex real-time RT-PCR assay for detection of influenza A, influenza B, RSV and typing of the 2009-H1N1 influenza virus

Development of a multiplex real-time RT-PCR assay for detection of influenza A, influenza B, RSV and typing of the 2009-H1N1 influenza virus

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