Development of a Real-Time Reverse Transcription-PCR Assay To Detect and Quantify Group A Rotavirus Equine-Like G3 Strains
| Autor: | Rashi Gautam, Michael D. Bowen, Mathew D. Esona, Eric M Katz |
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| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
Rotavirus Genotype Reverse Transcription Biology medicine.disease_cause Real-Time Polymerase Chain Reaction Group A Virology Rotavirus Infections Reverse transcription polymerase chain reaction Feces Real-time polymerase chain reaction TaqMan medicine Animals Humans RNA Viral Horses Forward primer Viral load Phylogeny |
| Zdroj: | J Clin Microbiol |
| ISSN: | 1098-660X |
| Popis: | Since 2013, group A rotavirus strains characterized as novel DS-1-like intergenogroup reassortant “equine-like G3” strains have emerged and spread across 5 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 227 to 2.3 × 10(9) 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 intergenogroup reassortant equine-like G3 strains with high sensitivity and specificity, faster turnaround time, and decreased cost. It 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. |
| Databáze: | OpenAIRE |
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