Journal
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
Volume 243, Issue -, Pages 36-45Publisher
ELSEVIER
DOI: 10.1016/j.ijfoodmicro.2016.11.022
Keywords
Hepatitis A virus; Norovirus; Digital RT-PCR; RT-qPCR; Soft berries; PCR inhibition
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Funding
- Coralie Coudray-Meunier
- ANSES
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Raw fruits may harbour many pathogens of public health concern including enteric viruses, which are the leading cause of foodborne outbreaks. Recently, consumption of soft berries has been associated with increasing reports of norovirus and hepatitis A virus outbreaks in Europe. Due to their low infectious doses and low concentrations in food samples, an efficient and sensitive analytical method is required for virus detection. In this study we explored two different ways to improve the reference method for the detection of enteric viruses in soft fruits (ISO/TS 15216-1; 15216-2): an additional purification step after RNA extraction; and the detection of enteric viral genome by an absolute quantification method (microfluidic digital RT-PCR). Both of these approaches led to an improvement of enteric virus detection in soft berries by greatly lowering PCR inhibition, raising viral extraction efficiencies and enabling validation of controls using pure RNA extracts. The PCR inhibitor removal step can be easily included in the routine method. Absolute quantification by digital RT-PCR may be a relevant alternative method to standardize quantification of enteric viruses in foodstuffs. (C) 2016 Elsevier B.V. All rights reserved.
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