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CC BY V4.0, Open Access
Journal article
Evaluation of Two Triplex One-Step qRT-PCR Assays for the Quantification of Human Enteric Viruses in Environmental Samples
Food and environmental virology, Vol.9(3), pp.342-349
2017
PMCID: PMC5548846
PMID: 28391510
Abstract
Human enteric viruses are responsible for waterborne and shellfish-associated disease outbreaks worldwide. Quantitative reverse transcription PCR (qRT-PCR) is often used to assess the health risks associated with shellfish and environmental water, but viral titres in sediments are less commonly investigated. In this study, we developed and validated two multiplex qRT-PCR assays for aquatic sediment and shellfish samples targeting viruses that are a common cause of gastroenteritis (norovirus GI, GII and hepatitis A virus), two emerging viruses (sapovirus and hepatitis E virus), along with mengovirus (MgV), which is often used as a sample process control for the assessment of RNA extraction efficiency. Singleplex and multiplex assays demonstrated comparable PCR efficiencies and gave reliable results over a wide concentration range. The multiplex assays showed remarkable sensitivity with a limit of detection of 1 RNA copy/A mu L nucleic acid extract for all target viruses and limits of quantification of 3-18 RNA copies/A mu L for the targeted human pathogenic viruses and 20-40 RNA copies/A mu L for MgV. The results demonstrated the veracity of multiplex qRT-PCR for the estimation of viral titres in sediment and shellfish, allowing the rapid assessment of viral infection risks associated with environments exposed to wastewater contamination.
Details
- Title
- Evaluation of Two Triplex One-Step qRT-PCR Assays for the Quantification of Human Enteric Viruses in Environmental Samples
- Authors/Creators
- Kata Farkas - Bangor UniversityDafydd E. Peters - Bangor UniversityJames E. McDonald - Bangor UniversityAlexis de Rougemont - Procédés Alimentaires et MicrobiologiquesShelagh K. Malham - Bangor UniversityDavey L. Jones - Bangor University
- Publication Details
- Food and environmental virology, Vol.9(3), pp.342-349
- Publisher
- Springer Nature
- Number of pages
- 8
- Grant note
- NE/M011577/1; NE/M010996/1 / Natural Environment Research Council; UK Research & Innovation (UKRI); Natural Environment Research Council (NERC) NE/M010996/1 / Food Standards Agency (FSA) under Environmental Microbiology and Human Health (EMHH) Programme NE/M011577/1; NE/M010996/1 / NERC; UK Research & Innovation (UKRI); Natural Environment Research Council (NERC) Natural Environment Research Council (NERC); UK Research & Innovation (UKRI)
- Identifiers
- 991005560427107891
- Copyright
- © 2017 The Authors
- Murdoch Affiliation
- Centre for Sustainable Farming Systems
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- International collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.246 Diarrheal Diseases
- 1.246.710 Enteric Viruses
- Web Of Science research areas
- Environmental Sciences
- Food Science & Technology
- Microbiology
- Virology
- ESI research areas
- Microbiology