Analytical performance of norovirus real-time RT-PCR detection protocols in Canadian laboratories.
J Clin Virol
; 50(2): 109-13, 2011 Feb.
Article
em En
| MEDLINE
| ID: mdl-21071266
ABSTRACT
BACKGROUND:
Noroviruses (NoVs) are the leading cause of infectious gastroenteritis worldwide. Real-time reverse transcription PCR (real-time RT-PCR) is the preferred method of NoV detection for the majority of testing laboratories. Although the accepted target region for molecular detection assays is the conserved ORF1/ORF2 junction, multiple variations have been published with differences in primers, probes, reagents, multiplexing, etc.OBJECTIVES:
We assessed the detection limit for GII.4 NoV real-time RT-PCR assays as well as the ability to detect the non-GII.4 NoV genotypes in each participating laboratory. STUDYDESIGN:
A panel of 25 RNA samples was circulated to 18 testing laboratories for comparison of their real-time RT-PCR procedures for NoV detection.RESULTS:
Multiple protocols with slight differences in reagents or conditions successfully detected 10 genome equivalents or fewer of NoV per reaction. Multiplex procedures were significantly associated (p=0.04) with false negative results, particularly for a GI.2 strain. Sensitive detection was associated with false positive results (p=0.03).CONCLUSIONS:
Overall, the data indicate that comparable results are produced under slightly different assay conditions.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Infecções por Caliciviridae
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Norovirus
/
Gastroenterite
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
País/Região como assunto:
America do norte
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article