PandemieGrippaleV1, Main, Merge, bibRecord, 003944

Rapid quantification of single-nucleotide mutations in mixed influenza A viral populations using allele-specific mixture analysis

Identifieur interne : 003944 ( Main/Merge ); précédent : 003943; suivant : 003945

Rapid quantification of single-nucleotide mutations in mixed influenza A viral populations using allele-specific mixture analysis

Auteurs : Cindy M. Liu [États-Unis] ; Elizabeth M. Driebe [États-Unis] ; James Schupp [États-Unis] ; Erin Kelley [États-Unis] ; Jack T. Nguyen [États-Unis] ; James J. Mcsharry [États-Unis] ; QINGMEI WENG [États-Unis] ; David M. Engelthaler [États-Unis] ; Paul S. Keim [États-Unis]

Source :

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

RBID : Pascal:10-0069675

Descripteurs français

Pascal (Inist)
- Analyse quantitative, Mutation, Allèle, Polymorphisme mononucléotide, Génotype, Typage, Réaction chaîne polymérase, Antiviral, Grippe A, Résistance, Microbiologie, Méthode.
Wicri :
- topic : Analyse quantitative.

English descriptors

KwdEn :
- Allele, Antiviral, Genotype, Influenza A, Method, Microbiology, Mutation, Polymerase chain reaction, Quantitative analysis, Resistance, Single nucleotide polymorphism, Typing.

Abstract

Monitoring antiviral resistance in influenza is critical to public health epidemiology and pandemic preparedness activities. Effective monitoring requires methods to detect low-level resistance and to monitor the change in resistance as a function of time and drug treatment. Resistance-conferring single-nucleotide mutations in influenza virus are ideal targets for such methods. In the present study, fives sets of paired TaqMan® allele-specific PCR (ASPCR) assays were developed and validated for quantitative single-nucleotide polymorphism (SNP) analysis. This novel method using ΔCt is termed allele-specific mixture analysis (ASMA) or FluASMA. The FluASMA assays target L26F, V27A, A30T, and S31N mutations in the A/Albany/1/98 (H3N2) M2 gene and H275Y mutation in the A/New Caledonia/20/99 (H1N1) NA gene and have a limit of quantification of 0.25-0.50% mutant. The error for % mutant estimation was less than 10% in all FluASMA assays, with intra-run ΔCt coefficient of variance (CoV) at ≤2% and inter-run ΔCt CoV at ≤5%. Results from the current study demonstrate that FluASMA is a highly sensitive and quantitative SNP analysis method, even for minor mutant components (<1%).

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Le document en format XML

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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Allele</term>
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<term>Microbiology</term>
<term>Mutation</term>
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<term>Resistance</term>
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<term>Réaction chaîne polymérase</term>
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<term>Grippe A</term>
<term>Résistance</term>
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<front><div type="abstract" xml:lang="en">Monitoring antiviral resistance in influenza is critical to public health epidemiology and pandemic preparedness activities. Effective monitoring requires methods to detect low-level resistance and to monitor the change in resistance as a function of time and drug treatment. Resistance-conferring single-nucleotide mutations in influenza virus are ideal targets for such methods. In the present study, fives sets of paired TaqMan® allele-specific PCR (ASPCR) assays were developed and validated for quantitative single-nucleotide polymorphism (SNP) analysis. This novel method using ΔCt is termed allele-specific mixture analysis (ASMA) or FluASMA. The FluASMA assays target L26F, V27A, A30T, and S31N mutations in the A/Albany/1/98 (H3N2) M2 gene and H275Y mutation in the A/New Caledonia/20/99 (H1N1) NA gene and have a limit of quantification of 0.25-0.50% mutant. The error for % mutant estimation was less than 10% in all FluASMA assays, with intra-run ΔCt coefficient of variance (CoV) at ≤2% and inter-run ΔCt CoV at ≤5%. Results from the current study demonstrate that FluASMA is a highly sensitive and quantitative SNP analysis method, even for minor mutant components (<1%).</div>
</front>
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<tree><country name="États-Unis"><region name="Arizona"><name sortKey="Liu, Cindy M" sort="Liu, Cindy M" uniqKey="Liu C" first="Cindy M." last="Liu">Cindy M. Liu</name>
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<name sortKey="Driebe, Elizabeth M" sort="Driebe, Elizabeth M" uniqKey="Driebe E" first="Elizabeth M." last="Driebe">Elizabeth M. Driebe</name>
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<name sortKey="Kelley, Erin" sort="Kelley, Erin" uniqKey="Kelley E" first="Erin" last="Kelley">Erin Kelley</name>
<name sortKey="Liu, Cindy M" sort="Liu, Cindy M" uniqKey="Liu C" first="Cindy M." last="Liu">Cindy M. Liu</name>
<name sortKey="Mcsharry, James J" sort="Mcsharry, James J" uniqKey="Mcsharry J" first="James J." last="Mcsharry">James J. Mcsharry</name>
<name sortKey="Nguyen, Jack T" sort="Nguyen, Jack T" uniqKey="Nguyen J" first="Jack T." last="Nguyen">Jack T. Nguyen</name>
<name sortKey="Qingmei Weng" sort="Qingmei Weng" uniqKey="Qingmei Weng" last="Qingmei Weng">QINGMEI WENG</name>
<name sortKey="Schupp, James" sort="Schupp, James" uniqKey="Schupp J" first="James" last="Schupp">James Schupp</name>
</country>
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</record>

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Rapid quantification of single-nucleotide mutations in mixed influenza A viral populations using allele-specific mixture analysis

Rapid quantification of single-nucleotide mutations in mixed influenza A viral populations using allele-specific mixture analysis

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Abstract

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