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The biotin-avidin-peroxidase (BAP) technique for measuring serum neutralizing (NT) antibody against rabies virus in human sera was established. When the focus reduction rates with serial serum dilutions were plotted on a probit chart, a linear regression line was drawn. The slopes (b-value) of the regression lines were identical irrespective of the different test samples. The sensitivity of the BAP test for detecting NT antibody was highest when compared with that of the rapid fluorescent focus inhibition test (RFFIT) and mouse neutralization test. In addition, Iow level-antibody titer (< I : 5) which could not be detected by mouse neutralization test was detected by the BAP test and the RFFIT. Serum NT antibody response was easily followed by the BAP test with individuals who received 3 doses of preexposure preventive rabies vaccine. Most of the vaccinees produced a detectable amount ofserum NT antibody by a single dose ofvaccine and after the second vaccination, all vaccinees produced high level of the antibody and the titers increased continuously until day 2 1 , however, the titers on day 78, two months after the last vaccination, showed a tendency ofdecline. INTRODUCTION NT antibody to rabies virus has been mostly measured by mouse neutralization test (Atanasiu, 1973) and rapid fluorescent focus inhibition test (RFFIT) (Smith et al., 1 973). The former method requires a large number of mice and at least 2 weeks of long period until the final results can be obtained. The latter method is much more superior to mouse neutralization test for detecting antibody in every respect of the sensitivity, reproducibility and rapidity. However, this method has a disadvantage that the fluorescent staining is unstable and the stained cells easily fade out and has a difficulty in testing many specimens collectively. In order to resolve this problem, we previously studied the biotin-avidin-peroxidase (BAP) technique for measurement of rabies virus infectivity and the NT antibody to rabies virus (Shichijo et al., 1 982). The BAP technique provides such advantages that the specimens can be examined under an ordinary microscope and can be stored for 1 Department of Microbiology, Medical College of Oita, Oita 87(~9 1 , Japan 2 The Chemo-SeroTherapeutic Research Institute, 668 Shimizu Machi, Kumamoto 860, Japan

nihon nettai igakkai zasshi/japanese journal of tropical medicine and hygiene

Sensitivity of the biotin-avidin-peroxidase (BAP) technique for rabies virus neutralizing antibody assay and the measurement of neutralizing antibodies of vaccinated human sera.