Application of an Ultracentrifugation-based Method for Detection of Feline Calicivirus (a Norovirus Surrogate) in Experimentally Contaminated Delicatessen Meat Samples

Abstract

The aim of this study was the application of an ultracentrifugation-based method for detection of feline calicivirus (FCV) in experimentally contaminated meat samples of roast pork chop, salami and gammon. Virus particles were liberated from food items by washing in 0.5-M glycine containing 1% bovine albumin. Food debris were removed by slow-speed centrifugation, and viruses were subsequently sedimented by ultracentrifugation. The recovery of infectious virus particles was determined by cell culture. Furthermore, RNA was extracted from virus particles and reverse-transcription polymerase chain reaction ((RT)PCR) was performed on the nucleic acid extracts. To demonstrate the efficiency of this method in removal of inhibitors from food sample extracts, an internal amplification control was constructed and incorporated into FCV (RT) PCR. Virus recovery for smoked meat, salami and cooked ham was 12.5%, 3.4% and 5.9%, respectively. The ultracentrifugation-based method is efficient in eliminating or reducing the level of inhibitory substances and can be applied for calicivirus extraction and concentration from delicatessen meat samples.

Evaluation of Extraction Methods for Efficient Detection of Enteric Viruses in Pork Meat Products

“…A key challenge for the detection of enteric viruses in food is that they can be present in low levels, and furthermore some pathogenic viruses, e.g., human NoV and HEV, are non-cultivatable or difficult to adapt for growth in vitro, so consequently their detection must be done directly from the food extracts, i.e., without any "enrichment" step in which the viral number can be increased (Croci et al 2008;Greening and Hewitt 2008;Rzeżutka et al 2008). Due to these constraints, the detection of enteric viruses in foods usually relies on molecular-based methods.…”

Porcine Blood and Liver as Sporadic Sources of Hepatitis E Virus (HEV) in the Production Chain of Offal-Derived Foodstuffs in Poland

“…It belongs to the virus collection of the Department of Food and Environmental Virology at the National Veterinary Research Institute in Poland. Preparation of the virus suspension in cell culture was previously described by Rzeżutka et al ( 2008 ).…”

Molecular detection of viruses in foods and food-processing environments

“…When viruses are extracted from fruits and vegetables, pectinase is frequently added to prevent jelly formation in the eluate by breaking the pectin bonds (Dubois et al ., 2002 ). When ultracentrifugation is used as concentration step, soya protein powder can be added in the purifi cation step to facilitate the liberation of viruses from food surfaces and thus improve recovery effi ciency (Rzeżutka et al ., 2006(Rzeżutka et al ., , 2008.…”

“…Viruses can be precipitated with ultracentrifugation from 100 000 × g to 235 000 × g (Casas et al ., 2007 ;Rutjes et al ., 2006a ;Rzeżutka et al ., 2006Rzeżutka et al ., , 2008. Virus eluates should be purifi ed prior to ultracentrifugation, either with high speed conventional centrifugation or 0.22-0.45 μ m-fi ltration, to remove food debris and other components which can co-sediment with virus particles during ultracentrifugation (Croci et al ., 2008 ).…”