An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping.
J Clin Virol
; 39(4): 318-21, 2007 Aug.
Article
en En
| MEDLINE
| ID: mdl-17604686
ABSTRACT
BACKGROUND:
Reverse transcription (RT)-PCR for norovirus detection is prone to false-negative results due to inhibitory substances in faeces. An internal control is needed to monitor extraction efficiency and to detect inhibition.OBJECTIVES:
To further develop a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS2 bacteriophage as an internal control. STUDYDESIGN:
Our norovirus RT-PCR assay was modified by addition of MS2 phage to the extraction tray and primers/probe for MS2 detection to the reaction mix. The effect of addition of MS2 phage and MS2 primers/probe on the sensitivity/specificity of the PCR assay was examined.RESULTS:
The addition of MS2 as an internal control showed no loss of sensitivity or specificity for norovirus detection.CONCLUSIONS:
A triplex, one-step, type-specific, real-time RT-PCR with MS2 internal control has been developed for use in routine laboratory diagnosis of norovirus infection.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
ARN Viral
/
Infecciones por Caliciviridae
/
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Norovirus
/
Gastroenteritis
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
J Clin Virol
Asunto de la revista:
VIROLOGIA
Año:
2007
Tipo del documento:
Article
País de afiliación:
Reino Unido