P2-99 Evaluation of a Novel Surface Sampling Wipe for Recovery of Human Noroviruses Prior to Detection Using RT-QPCR

Tuesday, July 30, 2013
Exhibit Hall (Charlotte Convention Center)
Grace Tung, North Carolina State University, Raleigh, NC
Caleb Wilson, North Carolina State University, Raleigh, NC
Lee-Ann Jaykus, North Carolina State University, Raleigh, NC
Arnaud Ganee, bioMérieux, Grenoble, France
Sylvanie Cassard, bioMérieux, Nantes, France
Claude Mabilat, bioMérieux, Grenoble, France
Introduction: Human noroviruses (NoV) are a significant cause of foodborne disease.  Lab-based and epidemiological studies demonstrate their high degree of environmental persistence and their resistance to commonly used surface sanitizers.  Environmental sampling is an important tool for assessing the effectiveness of cleaning and sanitizing regimens. 

Purpose: To test the efficacy of a novel surface sampling material (“wipes”) relative to human NoV recovery. 

Methods: Dilutions of a representative human NoV GII.4 outbreak strain  were applied to frequently used hard surfaces (stainless steel and ceramic) and the surface of representative foods (green pepper, apple, tomato, and cheese).  After inoculum drying, the sampler was used to recover viruses by wiping the surface in a circular, back and forth motion. The wipes were then directly processed for nucleic acid extraction using the automated EasyMag system, and the viral RNA quantified by RT-qPCR targeting the orf 1-orf 2 junction.  The efficiency of recovery of virus by the wipes was determined by direct comparison of RT-qPCR signals to a standard curve produced from the NoV GII.4 inoculum. 

Results: Recovery efficiency of the wipes ranged from 37-100% when used for sampling fresh fruits, vegetables, and hard surfaces artificially inoculated with high concentrations (3.4-5.4 log genome equivalent copies, GEC) of GII.4 human NoV.  At lower inoculum concentrations (1.4-2.4 log GEC), the wipes also yielded positive RT-qPCR signals for NoV.  Less efficient recovery was observed for cheese, ranging from 18-52% at high inoculum concentration, and failure to detect the GII.4 strain at lower concentrations.

Significance: The wipes are a user-friendly means by which to sample hard surfaces and foods that also provide high recovery efficiency for human NoV.  They are being field tested in environmental surveys designed to evaluate NoV prevalence in public restrooms during the typical epidemic (winter) season.