An in situ transcription method was developed to detect hepatitis A virus RNA in both cell cultures and shellfish tissues. Radiolabeled cDNA copies were synthesized in situ by reverse transcriptase-directed transcription after annealing with a specific primer to the viral RNA. Both tritium (3H) and 35S were useful in the in situ transcription reaction, but the use of 3H resulted in a lower background and finer detail in the localization of viral particles. Application of the method to different organs of oysters which had bioaccumulated hepatitis A virus allowed the first in situ localization of the virus, specifically in stomach and hepatopancreatic tissues.
During 1981–1982, epidemics of acute hemorrhagic conjunctivitis (AHC) occurred in and around Chandigarh, India. Conjunctival swabs were taken from AHC cases and investigated by the indirect immunofluorescence method both in Chandigarh and in Houston. By using reference antiserum, both laboratories proved enterovirus type 70 to be the causative agent of the epidemics. The immunofluorescence test was evaluated as a rapid method for the diagnosis of AHC.
Sera of 25 patients with membranous glomerulonephritis (MGN), 16 with membranoproliferative glomerulonephritis (MPGN) and 54 with IgA glomerulonephritis (IgA GN) were studied for complement-binding antibodies to herpes simplex virus (HSV), cytomegalovirus (CMV) and antibodies to various Epstein-Barr virus (EBV) associated antigens, as also for the titres of these antibodies. The sera of 220 normal individuals served as controls, 120 controls being used for each case. Anti-HSV titres of greater than or equal to 1:64 were found to occur in the sera of all three GN groups in a higher proportion than in those of the controls. This was also valid for the complement-binding antibodies to CM, although here the differences were not invariably significant. IgA antibodies reacting with EBV capside antigen (EBVCA) were likewise of statistically increased frequency in IgA GN, as also in MPGN, and in these two groups the geometric mean of the reciprocal value of the IgA antibody titres was also higher than either in the controls or in MGN. The results of the studies carried out within 6 months after onset of renal disease point to an EBV infection, either fresh or having taken place in the recent past, in 20 cases. These data are compatible with a direct or indirect role of EBV in the production and/or persistence of certain types of GN. The high anti-HSV and CMV titres suggest that in a number of patients with renal disease the immune responses to certain types of the HSV group may be abnormal.
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