ArticleBMC infectious diseases2016
A high throughput serum bactericidal assay for antibodies to Haemophilus influenzae type b.
Article in BMC infectious diseases, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 12 citations in OpenAlex.
- Preclinical development of a well-defined semisynthetic glycoconjugate vaccine againstScience advances · 2026Article
- Serum susceptibility of Escherichia coli and its association with patient clinical outcomes.PloS one · 2024Article
- Increasing the High Throughput of a Luminescence-Based Serum Bactericidal Assay (L-SBA).Biotech (Basel (Switzerland)) · 2021Article
- Evaluating Functional Immunity Following Encapsulated Bacterial Infection and Vaccination.Vaccines · 2021Article
- Antibody Binding and Complement-Mediated Killing of Invasive Haemophilus influenzae Isolates from Spain, Portugal, and the Netherlands.Infection and immunity · 2020Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe protective capacities of antibodies induced with Haemophilus influenzae type b (Hib) vaccines can be directly assessed in vitro with a Hib-specific serum bactericidal assay (SBA). However, the conventional SBA requires several tedious steps including manual counting of bacterial colonies, and therefore, it is seldom used.
methodsTo overcome these limitations, we have improved the conventional SBA by using frozen target bacteria and by developing an automated colony counting method based on agar plates with the chromogenic dye 2, 3, 5-triphenyl tetrazolium chloride (TTC).
resultsThese changes enabled us to analyze about 100 serum samples per day per person by SBA. When the intra- and inter-assay precisions were studied, this assay showed a coefficient of variation (CV) ranging from 1 to 38 %. To monitor the long term assay stability for assays involving different bacteria lots, complement lots, and operators, we analyzed bactericidal indices of quality control samples obtained over a 6 year period and found the CV to be about 35-50 %. Lastly, our SBA results were compared with the ELISA results obtained using 90 serum samples from children. We showed that the bactericidal index correlated with IgG anti-Hib antibody levels (r = 0.84), with a bactericidal index of 10 corresponding approximately to 0.15 μg/mL IgG, the widely accepted protective level of antibody.
conclusionWe describe a simple high throughput SBA for anti-Hib antibodies that would be useful for evaluating various Hib vaccines. While additional work will be needed to standardize the assay, this SBA should greatly facilitate studies of Hib vaccines.
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