Development of a Real-Time Reverse Transcription-PCR Assay To Detect and Quantify Group A Rotavirus Equine-Like G3 Strains
- 19 October 2021
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 59 (11)
- https://doi.org/10.1128/jcm.02602-20
Abstract
Since 2013, group A rotavirus strains characterized as novel DS-1-like inter-genogroup reassortant ‘equine-like G3’ strains have emerged and spread across five continents among human populations in at least 14 countries. Here we report a novel one-step TaqMan quantitative real-time reverse transcription-PCR assay developed to genotype and quantify the viral load for samples containing rotavirus equine-like G3 strains. Using a universal G forward primer and a newly designed reverse primer and TaqMan probe, we developed and validated an assay with a linear dynamic range of 2.3 × 109 – 227 copies per reaction and a limit of detection of 227 copies. The percent positive agreement, percent negative agreement, and precision of our assay were 100.00%, 99.63%, and 100.00%, respectively. This assay can simultaneously detect and quantify the viral load for samples containing DS-1-like inter-genogroup reassortant equine-like G3 strains with high sensitivity and specificity, faster turnaround time, and decreased cost and will be valuable for high-throughput screening of stool samples collected to monitor equine-like G3 strain prevalence and circulation among human populations throughout the world.Keywords
Funding Information
- HHS | Centers for Disease Control and Prevention
This publication has 46 references indexed in Scilit:
- Highly sensitive detection of the group A Rotavirus using Apolipoprotein H-coated ELISA plates compared to quantitative real-time PCRVirology Journal, 2011
- Novel Light-Upon-Extension Real-Time PCR Assay for Simultaneous Detection, Quantification, and Genogrouping of Group A RotavirusJournal of Clinical Microbiology, 2010
- Evolutionary Dynamics of Human Rotaviruses: Balancing Reassortment with Preferred Genome ConstellationsPLoS Pathogens, 2009
- The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR ExperimentsClinical Chemistry, 2009
- Whole genome sequence and phylogenetic analyses reveal human rotavirus G3P[3] strains Ro1845 and HCR3A are examples of direct virion transmission of canine/feline rotaviruses to humansVirology, 2008
- Sensitive Detection of Multiple Rotavirus Genotypes with a Single Reverse Transcription-Real-Time Quantitative PCR AssayJournal of Clinical Microbiology, 2008
- Eleven Golden Rules of Quantitative RT-PCRTHE PLANT CELL ONLINE, 2008
- Recommendations for the classification of group A rotaviruses using all 11 genomic RNA segmentsArchiv für die gesamte Virusforschung, 2008
- Full Genome-Based Classification of Rotaviruses Reveals a Common Origin between Human Wa-Like and Porcine Rotavirus Strains and Human DS-1-Like and Bovine Rotavirus StrainsJournal of Virology, 2008
- Real-Time Reverse Transcription-PCR for Detection of Rotavirus and Adenovirus as Causative Agents of Acute Viral Gastroenteritis in ChildrenJournal of Clinical Microbiology, 2006