ArticleFood and environmental virology2017
Improved Detection of Norovirus and Hepatitis A Virus in Surface Water by Applying Pre-PCR Processing.
Article in Food and environmental virology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Wastewater-based surveillance as a tool for public health action: SARS-CoV-2 and beyond.Clinical microbiology reviews · 2024Review
- Surfactant Treatment for Efficient Gene Detection of Enteric Viruses and Indicators in Surface Water Concentrated by Ultrafiltration.Food and environmental virology · 2023Article
- Comparison of Skimmed Milk and Lanthanum Flocculation for Concentration of Pathogenic Viruses in Water.Food and environmental virology · 2021Article
- Microbial Indicators of Fecal Pollution: Recent Progress and Challenges in Assessing Water Quality.Current environmental health reports · 2020Review
- PCR inhibition in qPCR, dPCR and MPS-mechanisms and solutions.Analytical and bioanalytical chemistry · 2020Review
- Interlaboratory Comparative Study to Detect Potentially Infectious Human Enteric Viruses in Influent and Effluent Waters.Food and environmental virology · 2019Article
- Missing the Match Might Not Cost You the Game: Primer-Template Mismatches Studied in Different Hepatitis A Virus Variants.Food and environmental virology · 2019Article
- HEV Occurrence in Waste and Drinking Water Treatment Plants.Frontiers in microbiology · 2019Article
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) detection of waterborne RNA viruses generally requires concentration of large water volumes due to low virus levels. A common approach is to use dead-end ultrafiltration followed by precipitation with polyethylene glycol. However, this procedure often leads to the co-concentration of PCR inhibitors that impairs the limit of detection and causes false-negative results. Here, we applied the concept of pre-PCR processing to optimize RT-qPCR detection of norovirus genogroup I (GI), genogroup II (GII), and hepatitis A virus (HAV) in challenging water matrices. The RT-qPCR assay was improved by screening for an inhibitor-tolerant master mix and modifying the primers with twisted intercalating nucleic acid molecules. Additionally, a modified protocol based on chaotropic lysis buffer and magnetic silica bead nucleic acid extraction was developed for complex water matrices. A validation of the modified extraction protocol on surface and drinking waters was performed. At least a 26-fold improvement was seen in the most complex surface water studied. The modified protocol resulted in average recoveries of 33, 13, 8, and 4% for mengovirus, norovirus GI, GII, and HAV, respectively. The modified protocol also improved the limit of detection for norovirus GI and HAV. RT-qPCR inhibition with C
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Registered trials
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