Abstract
COLLEAGUES -During recent years several antibody assays for hepatitis-A virus (HAV) have been developed. These assays detect the following different classes of serum antibodies: IgGclass (anti-HAV IgG), IgM-class (anti-HAV IgM), and IgAclass (anti-HAV IgA). Neutralizing antibody to HAV (antiHAV Nt) has not been detected previously, but we have developed and here report a microtiter test system for detecting it. This anti-HAV Nt assay, which uses fetal rhesus-monkey kidney (Frhk)-4/R cells and the HAV strain GBM/Frhk-4/R that has been adapted to grow in these cells [1], was performed as follows: samples of sera to be tested were diluted in Earle's minimal essential medium (Serva Fein Biochemica, Heidelberg, FRG) containing 5% fetal calf serum (Flow Laboratories, Bonn, FRG) and 2.5% inactivated anti-HAV-negative human serum and inactivated by heating at 56 C for 30 min. Equal volumes of serum dilutions and of HAV suspensions containing 5,000 TC113,0 were mixed and incubated at 37 C for 4 hr. Thereafter, each microtiter well with Frhk-4/R cells received 0.2 ml of the virus/serum mixture. Different virus titrations and sam-
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