ENZYMATIC TREATMENT OF FORMALIN-FIXED AND PARAFFIN-EMBEDDED SPECIMENS FOR DETECTION OF ANTIGENS OF HERPES-SIMPLEX, VARICELLA-ZOSTER AND HUMAN CYTOMEGALOVIRUSES

  • 1 January 1982
    • journal article
    • research article
    • Vol. 52  (1) , 17-25
Abstract
Attempts were made to detect antigens of herpes simplex (HS), varicella-zoster (VZ) and human cytomegalo (CM) viruses in pathological preparations made by formalin fixation and paraffin embedding and in infected culture cells by means of enzymatic treatment followed by indirect immunofluorescent staining. Specimens from autopsy cases, in which diagnosis was established by detection of viral antigens or virus isolation, showed specific antigens in specimens of various organs of all cases of HS (6 cases), VZ (3 cases) and CM (2 cases). In 17 other cases which were suspected for those viral infections on the basis of ordinary light microscopic observation, respective viral antigens were also specifically demonstrated, enabling differential diagnosis. In 1 case, HS antigen was detected in the esophagus and CM antigen in the liver, testifying to the occurrence of a double infection. The time span during which the specific antigens could be preserved was tested and preservation of stainable antigens was better in paraffin-embedded specimens than in specimens kept in formalin. HS and VZ antigens were detected in paraffin-embedded specimens prepared 20 and 18 yr ago, respectively. The optimal conditions of trypsin treatment varied depending on the organ and on individual specimen, but a commonly recommendable procedure was to warm at 37.degree. C for 3 h with 0.25% trypsin. A test with HS virus-infected African green monkey kidney Vero cells indicated that the effect of trypsin treatment was influenced by the concentration of formalin and the time of fixation.