MAGNETIC IMMUNOSEPARATION PCR ASSAY (MIPA) FOR DETECTION OF HEPATITIS-A VIRUS (HAV) IN AMERICAN OYSTER (CRASSOSTREA-VIRGINICA)

In order to detect the low numbers of hepatitis A viral (HAV) particle s which map potentially be present in food and cause a serious illness , an original procedure which combines immunomagnetic separation and P CR is described. The use of streptavidin magnetic beads coated with bi otinylated human anti-HAV IgG allows virus capture and the removal of the RT-PCR inhibitory compounds which usually are present in shellfish extracts. Following immunomagnetic capture, the separated HAV were ly sed, the beads discarded, and the supernatant containing the viral RNA subjected to the RT-PCR protocol. Levels of HAV ranging from 10 to 10 (5) pfu were successfully detected in artificially contaminated sample s of shucked American oyster (Crassostrea virginica).