IN-SITU DETECTION OF HEPATITIS-A VIRUS IN CELL-CULTURES AND SHELLFISHTISSUES

An in situ transcription method was developed to detect hepatitis A vi rus RNA in both cell cultures and shellfish tissues. Radiolabeled cDNA copies were synthesized in situ by reverse transcriptase-directed tra nscription after annealing with a specific primer to the viral RNA. Bo th tritium (H-3) and S-35 were useful in the in situ transcription rea ction, but the use of H-3 resulted in a lower background and finer det ail in the localization of viral particles. Application of the method to different organs of oysters which had bioaccumulated hepatitis A vi rus allowed the first in situ localization of the virus, specifically in stomach and hepatopancreatic tissues.