Molecular genotyping and quantitation assay for rotavirus surveillance.
J Virol Methods
; 213: 157-63, 2015 Mar.
Article
en En
| MEDLINE
| ID: mdl-25526999
ABSTRACT
Rotavirus genotyping is useful for surveillance purposes especially in areas where rotavirus vaccination has been or will be implemented. RT-PCR based molecular methods have been applied widely, but quantitative assays targeting a broad spectrum of genotypes have not been developed. Three real time RT-PCR panels were designed to identify G1, G2, G9, G12 (panel GI), G3, G4, G8, G10 (panel GII), and P[4], P[6], P[8], P[10], P[11] (panel P), respectively. An assay targeting NSP3 was included in both G panels as an internal control. The cognate assays were also formulated as one RT-PCR-Luminex panel for simultaneous detection of all the genotypes listed above plus P[9]. The assays were evaluated with various rotavirus isolates and 89 clinical samples from Virginia, Bangladesh and Tanzania, and exhibited 95% (81/85) sensitivity compared with the conventional RT-PCR-Gel-electrophoresis method, and 100% concordance with sequencing. Real time assays identified a significantly higher rate of mixed genotypes in Bangladeshi samples than the conventional gel-electrophoresis-based RT-PCR assay (32.5% versus 12.5%, P<0.05). In these mixed infections, the relative abundance of the rotavirus types could be estimated by Cq values. These typing assays detect and discriminate a broad range of G/P types circulating in different geographic regions with high sensitivity and specificity and can be used for rotavirus surveillance.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Infecciones por Rotavirus
/
Rotavirus
/
Carga Viral
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Técnicas de Genotipaje
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Monitoreo Epidemiológico
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
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Screening_studies
Límite:
Child, preschool
/
Humans
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Infant
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Newborn
País/Región como asunto:
Africa
/
America do norte
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Asia
Idioma:
En
Revista:
J Virol Methods
Año:
2015
Tipo del documento:
Article