Microneutralization test for influenza A and B and parainfluenza 1 and 2 viruses that uses continuous cell lines and fresh serum enhancement.

A modified microneutralization test for influenza A and B and parainfluenza 1 and 2 viruses was developed. Use of continuous cell lines (Madin-Darby canine kidney and LLC-MK2), hemagglutination for virus detection, and transfer plates and other microtiter equipment resulted in a highly mechanized technique suitable for titrating large numbers of sera easily and relatively inexpensively. Titers of heat-inactivated human sera were enhanced 0.5 to 3.5 log2 by addition of fresh hamster or human serum to the test. Seroconversion rates and identification of seronegative persons were not changed by fresh serum enhancement, but the magnitude of seroconversion was often increased and the distribution of positive titers was broadened. For influenza A and B, seroconversion rates in the microneutralization test were equivalent to those obtained in rhesus monkey kidney tube neutralization tests. For influenza B, seroconversion rates by micro-neutralization were almost double those obtained with the hemagglutination inhibition test, but the rates were similar for influenza A/USSR (H1N1). Preexisting microneutralization titers correlated well with resistance of infection with influenza B. Limited experience with parainfluenza 1 and 2 was similar to previously reported findings with the tube neutralization test.