DIN CEN ISO/TS 15216-1:2013-08

Hepatitis A virus (HAV) and norovirus (NoV) are important agents of food-borne human viral illness. No routine methods exist to culture these viruses from food matrices. Detection is therefore reliant on molecular methods using the reverse-transcriptase polymerase chain reaction (RT-PCR). As many food matrices contain substances that are inhibitory to RT-PCR, it is necessary to use an extraction method that produces highly clean RNA preparations that are fit for purpose. For all matrices which are not covered by this Technical Specification, it is necessary to validate this method. This Technical Specification describes a method for quantification of levels of HAV and NoV genogroup I (GI) and II (GII) RNA, from test samples of foodstuffs or food surfaces. Following liberation of viruses from the test sample, viral RNA is then extracted by lysis with guanidine thiocyanate and adsorption on silica. Target sequences within the viral RNA are amplified and detected by real-time RT-PCR. This approach is also relevant for detection of the target viruses on fomites, or of other human viruses in foodstuffs, on food surfaces or on fomites following appropriate validation and using target-specific primer and probe sets. This Technical Specification is valid for consumer health and protection. The committee responsible for this Technical Specification is NA 057-01-06-20 AK "Viren" ("Viruses") at DIN.