Rapid quantification of single-nucleotide mutations in mixed influenza A viral populations using allele-specific mixture analysis.
J Virol Methods
; 163(1): 109-15, 2010 Jan.
Article
em En
| MEDLINE
| ID: mdl-19761797
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 Delta Ct 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 Delta Ct coefficient of variance (CoV) at
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Polimorfismo de Nucleotídeo Único
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Farmacorresistência Viral
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Influenza Humana
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Vírus da Influenza A Subtipo H1N1
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Vírus da Influenza A Subtipo H3N2
Limite:
Humans
Idioma:
En
Revista:
J Virol Methods
Ano de publicação:
2010
Tipo de documento:
Article
País de afiliação:
Estados Unidos