A passive haemagglutination test for rabies antibodies using rabies glycoprotein coupled erythrocytes.

The presently recommended tests for assaying rabies antibodies like mouse neutralization test (MINT) and rapid fluorescent focus inhibition test (RFFIT) are either time consuming or expensive and are generally performed in reference laboratories. There is a need to develop a specific and rapid method for detection of rabies antibodies that can be used to monitor sero-conversion after pre-or post-exposure vaccination. In this study, we have developed a passive haemagglutination (PHA) using purified rabies virus glycoprotein coupled to sheep erythrocytes using chromium chloride (0.04%) as a coupling agent. Two hundred and fifty five serum samples from people vaccinated with different rabies vaccines, 16 paired serum and CSF samples from autopsy confirmed cases of paralytic rabies, and serum samples from 65 normal healthy controls were tested and evaluated in comparison to standard MNT. Among the vaccinees, 250 samples were positive both by MNT and PHA but 5 samples were negative by PHA and positive by MNT. The titres obtained by PHA were lower compared to MNT, but there was significant correlation between the two (r=0.885). The specificity of the test was 99.7% and sensitivity was 100% as compared to MNT. Thus this PHA test promises to be a rapid and specific test for assaying rabies antibodies and may be useful in screening large number of serum samples for sero conversion after vaccination. It may also assist in rapid laboratory confirmation of paralytic rabies cases, based on detection of antibodies in CSF and serum.

Comparative evaluation of a simple indirect immunofluorescence test and mouse neutralization test for assaying rabies antibodies.

In this study, we have developed and evaluated a simple indirect immunofluorescence test (IIFT) to detect rabies antibodies in a two-step immunofluorescence assay. One hundred and eighty five serum samples from people who had taken different rabies vaccines and 8 pairs of serum and CSF samples from confirmed paralytic rabies cases were tested by IIFT and results evaluated in comparison to standard mouse neutralization test (MNT). Though the titres of rabies antibodies obtained with IIFT were 2-4 times lesser in comparison to MNT, a significant correlation was seen between the two tests (R = 0.883). The specificity of this IIFT was found to be 97.9% and the sensitivity was 97.2%. These results indicate that this simple and rapid IIFT can be used to screen large number of serum samples to monitor sero-conversion after pre or post exposure vaccination and may also assist in rapid ante-mortem diagnosis of atypical human rabies.