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A sensitive method for demonstrating the site of hepatitis B surface antigen (HBsAg) in fixed tissues embedded in either paraffin or araldite is described. The method employs the peroxidase-rabbit antiperoxidase linkage through goat antirabbit to rabbit anti-HBsAg. In staining hepatitis antigen in agar, comparison of fixation (using three common fixatives) with unfixed precipitation arcs revealed no recognizable differences in antigenicity induced by fixation. The method allows confirmation of positive reaction by appropriate blocking controls. The technic is compared with the orcein stain of Shikata and found to be somewhat more sensitive but slightly more time-consuming.

Two hundred seventy-nine patients who died of hepatocellular carcinoma were autopsied at Los Angeles County--USC Medical Center and the John Wesley--USC Liver Unit from 1949 through 1974, and tissues from 168 of these cases were available for staining for hepatitis B surface antigen (HBSAg). Twenty-one per cent of the livers had stainable HBSAg. There were prominent increases both in total numbers of hepatic cancers and in the percentages that were HBSAg-positive beginning about 1970, but the numbers of hepatocellular carcinomas arising in noncirrhotic livers also increased. From 1969 to 1974, 73% of those who had hepatocellular carcinomas arising to nonalcoholic but cirrhotic livers were HBSAg-positive. Racial differences in the incidences of cirrhosis, the incidences of hepatocellular carcinomas associated with HBSAg were found. The incidences of cirrhosis were: Caucasian 11%; Mexican 12.2%; Negro 5.7:; Oriental 10%. Hepatocellular carcinomas arose in 3.2% of Caucasians who had cirrhosis; 3.6% of Mexicans; 8.3% of Negroes; 47% or Orientals. Ten per cent of Caucasians who had hepatocellular carcinomas in cirrhotic livers were HBSAg-positive; 25% of Negroes; 12% of Mexicans; 47% of Orientals.