Major achievements in REFHEPA

Development of standard reference methods for hepatitis A virus and Norovirus in bivalve molluscan shellfish

Following the comparison of two candidate real-time PCR methodologies for the detection of HAV a real-time one-step RT-TaqMan PCR method developed by the University of Barcelona has been selected for validation among partners. Validation is currently being completed by partners by testing shellfish and clinical samples. Approaches to standardisation being developed include the addition of a murine virus (Mengo virus) to the sample to determine efficiency of the extraction procedure. A real-time one-step RT-TaqMan PCR for Mengo virus has been developed for this purpose. The addition of a known concentration of ssRNA control molecules to allow the efficiency of the RT-PCR step is also being validated. This approach will allow the development of a standardised HAV method.

A real-time RT-TaqMan PCR detection method for genogroup II NoV has been developed and validated in spiked samples of shellfish by IFREMER and the University of Helsinki. Validation of this method continues using naturally contaminated shellfish. Broadly reactive primers and probes for detecting genogroup II NoV have been developed and assessed. The developed assay is currently being validated and initial work in spiked samples demonstrates increased performance over other candidate primer probe sets.

People in this project