Comparison of hemagglutination inhibition, single radial hemolysis, virus neutralization assays, and ELISA to detect antibody levels against seasonal influenza viruses

Abstract

BackgroundThe immunological response to influenza vaccine and/or natural infection is evaluated by serological techniques, the most common being hemagglutination inhibition (HI), single radial hemolysis (SRH), and virus neutralization assays, which is commonly used in a micro‐neutralization (MN) format. ELISA is not officially required; however, this assay is able to measure different class‐specific antibodies. The four assays identify different sets or subsets of antibodies.ObjectivesThe aim of this study was to establish the correlation among four serological assays using four seasonal influenza strains.MethodsThe HI, SRH, MN assays, and ELISA were performed on four seasonal influenza strains.ResultsA strong positive correlation was found between HI and MN and between SRH and MN assays for influenza A strains. The B strains also showed good correlations among the three assays. A positive correlation was also found between ELISA and the “classical” assays for all strains. Concerning the correlates of protection, as defined by HI ≥ 40 and SRH ≥ 25 mm2, good agreement was observed for the influenza A strains. By contrast, the agreement for the B strains was very low.ConclusionsThere is a positive strong correlation among the four serological assays for both A and B strains, especially for the HI and MN assays. There is good agreement on correlates of protection between HI and SRH assays for the A strains, but very low agreement for the B strains, suggesting higher sensitivity of SRH than HI assay in detecting antibodies against the influenza B viruses.

Influenza, Malaria parasitaemia and Typhoid fever Coinfection in Children: Seroepidemiologic Investigation in Four Healthcare Centres in Lagos, Nigeria

“…Enzyme linked immunosorbent assay (ELISA) is a sensitive and quantifiable diagnostic method that has been used in the detection of different aetiologies [22]. Sera antibodies are often used to correlate influenza detection in the assay [23]. This study detected a total percentage seroprevalence of 20.9% to influenza virus among paediatric patients attending different healthcare centers in Lagos, Nigeria.…”

The use of cell-mediated immunity for the evaluation of influenza vaccines: an upcoming necessity

“…26 HI also presents limitations, including low sensitivity for influenza B and avian viruses, unsuitability for LAIV evaluation, high inter-laboratory variability due to many factors and the absence of standardized protocols. 25 Regarding seasonal influenza A strains, SRH and HI show similar sensitivity. 24 A ≥ 25 mm 2 zone is defined as a correlate of protection for SRH.…”

“…Hence, MN can detect a higher proportion of protective antibodies than SRH and HI. 25 Specifically, HI has been seen to have lower sensitivity than SRH and MN, as demonstrated by the fact that 34% and 16% of the subjects, respectively, showed HI and SRH titers below the detection limit, in comparison with 7% on the MN assay. Previous investigations had also reported a higher sensitivity of MN than HI.…”

“…22,23 The SRH assay has a higher sensitivity for influenza B strains. 24,25 The HI assay is the gold-standard assay of antibody titers against HA, and is based on erythrocyte agglutination due to the ability of antibodies that specifically recognize HA to inhibit the binding of viral surface protein HA to sialic acid sites on the surface of red blood cells. 21 Avian (chicken or turkey) or mammalian (horse or guinea pig) erythrocytes are usually chosen for the assay.…”

Serologically-based Evaluation of Cross-protection Antibody Responses Among Different A(H1N1) Influenza Strains

“…The immunological response to influenza vaccine and/or natural infection is mainly evaluated by two serological techniques, the haemagglutination inhibition (HI) and the single radial haemolysis (SRH) which still are the most widely used and officially recognized serological assays by regulatory authorities [16]. In recent years, the importance of neutralization assays for serological evaluations has been increased and the analysis of neuraminidase-specific antibodies has also become a topic of increasing interest [17].…”