A multiplex one-step real-time RT-PCR assay for influenza surveillance.
Euro Surveill
; 16(7)2011 Feb 17.
Article
em En
| MEDLINE
| ID: mdl-21345319
ABSTRACT
For surveillance purposes real-time PCR assays for influenza viruses had to be adapted to the pandemic influenza A(H1N1)2009 strain. We combined published primers and probes for influenza A, influenza B and an internal amplification control with a detection system for influenza A(H1N1)2009 to set up a rapid, reliable, simple and cost-effective high-throughput multiplex one-step real-time RT-PCR. The workflow also includes automated sample preparation for high-throughput screening. The lower limit of detection of the multiplex assay was 3.5x10(2) RNA copies per PCR reaction. The diagnostic sensitivity of the multiplex assay was 87.7%, but increased to 99.4% for influenza-positive samples yielding C(t) values of less than 34 cycles in the respective diagnostic assay. High specificity was confirmed by sequencing and correct detection of 15 reference samples from two quality assurance studies. The multiplex PCR was introduced for surveillance of samples from a network of general practitioners and paediatricians in Bavaria, Germany during the influenza pandemic of 2009. Comparison with surveillance data from reported cases proved the reliability of the multiplex assay for influenza surveillance programmes.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Vírus da Influenza A
/
Vírus da Influenza B
/
Vigilância da População
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Influenza Humana
/
Vírus da Influenza A Subtipo H1N1
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Limite:
Humans
País/Região como assunto:
Europa
Idioma:
En
Revista:
Euro Surveill
Assunto da revista:
DOENCAS TRANSMISSIVEIS
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Alemanha