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Single-step RT-PCR assay for dual genotyping of GI and GII norovirus strains.
Chhabra, Preeti; Browne, Hannah; Huynh, Thalia; Diez-Valcarce, Marta; Barclay, Leslie; Kosek, Margaret N; Ahmed, Tahmeed; Lopez, Maria Renee; Pan, Chao-Yang; Vinjé, Jan.
Afiliación
  • Chhabra P; Viral Gastroenteritis Branch, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA. Electronic address: pchhabra@cdc.gov.
  • Browne H; National Foundation for the Centers for Disease Control and Prevention Inc., Atlanta, GA, USA.
  • Huynh T; California Department of Public Health, Richmond, CA, USA.
  • Diez-Valcarce M; Rollins School of Public Health, Emory University, Atlanta, GA, USA.
  • Barclay L; Viral Gastroenteritis Branch, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
  • Kosek MN; University of Virginia Division of Infectious Diseases and International Health, Charlottesville, VA, USA.
  • Ahmed T; International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
  • Lopez MR; Universidad del Valle de Guatemala, Guatemala City, Guatemala.
  • Pan CY; California Department of Public Health, Richmond, CA, USA.
  • Vinjé J; Viral Gastroenteritis Branch, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
J Clin Virol ; 134: 104689, 2021 01.
Article en En | MEDLINE | ID: mdl-33260046
BACKGROUND: Noroviruses are the major cause of acute gastroenteritis (AGE) in people of all ages globally. Standardized genotyping is key for outbreak investigations and surveillance networks. OBJECTIVE: Here we describe the validation of a one-step conventional RT-PCR assay for sequence-based dual typing of GI and GII noroviruses. This polymerase (P) and capsid (C) dual typing assay uses a combination of previously published oligonucleotide primers amplifying a genomic region spanning the 3'-end of ORF1 and 5'end of ORF2 resulting in a 579 bp product for GI and 570 bp product for GII viruses. RESULTS: The limit of detection of the assay ranged from 5 to 50 copies of viral RNA per reaction for GI and GII. To validate the assay, we tested 2,663 noroviruspositive stool samples from outbreaks and sporadic cases of AGE in Bangladesh, Guatemala, Peru, and USA collected between 2010-2019, of which 2,392 (90 %) were genotyped successfully. Most of the known genotypes infecting humans (GI (n = 9) and GII (n = 23)) and P types (GI (n = 15), GII, (n = 20)) could be detected. The remaining 270 samples had low viral load (Ct > 30) by real-time RT-PCR. A panel of 166 samples positive for other enteric viruses (rotavirus, astrovirus, sapovirus, adenovirus type 40/41) tested negative. CONCLUSION: The use of broadly reactive genotyping assays greatly strengthens exchange of standardized genotype data globally to monitor trends in genotype diversity which is important for both the development of vaccines and to measure their impact.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones por Caliciviridae / Norovirus / Gastroenteritis Límite: Humans Idioma: En Revista: J Clin Virol Asunto de la revista: VIROLOGIA Año: 2021 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones por Caliciviridae / Norovirus / Gastroenteritis Límite: Humans Idioma: En Revista: J Clin Virol Asunto de la revista: VIROLOGIA Año: 2021 Tipo del documento: Article Pais de publicación: Países Bajos