Comparison of influenza serological techniques by international collaborative study

An international collaborative study was performed to investigate the reproducibility of influenza serological techniques. Participants in seven laboratories representing five countries measured antibody to A/Sichuan/2/87 (H3N2), A/Taiwan/1/86 (H1N1) and B/Beijing/1/87 influenza viruses in 11 human sera and three postinfection ferret sera. Two different serological techniques were used, haemagglutination inhibition (HI) and single-radial haemolysis (SRH) and, although each technique was reproducible within laboratories, variability between laboratories was higher for HI (maximum variability 32-fold; geometric coefficient of variation, GCV, 112%) than for SRH (maximum variability 3.8-fold; GCV 57%). The use of a standard serum allowed direct comparison of HI and SRH data and, for each technique, a standard serum improved inter-laboratory agreement. For influenza A viruses there was a correlation between HI and SRH antibodies (correlation coefficient approximately 0.9). An HI titre of 1:40 in human sera corresponded to an SRH titre of 19-33 mm2. The results of the study indicate that two sera would be expected to contain different antibody levels if their HI titres differed by > fourfold and SRH areas differed by > 50%. Both SRH and HI possessed equivalent sensitivity for measurement of antibody to influenza A viruses but SRH was more sensitive for detection of antibody to influenza B viruses. The study provided valuable information about standardization of antibody assays.