The Performance of Multiplex Immunoassays for Antibody Determination to Diphtheria, Tetanus and Pertussis: A Need for Standardisation

Abstract

Serological surveillance is a powerful method to estimate the risk of infectious diseases and their vaccination coverage among populations. However, enzyme-linked immunosorbent assays commonly used for antibody screening are time consuming when testing multiple analytes on large sample numbers. As multiplex immunoassays for detection of antibodies against tetanus, diphtheria and pertussis (or other vaccine-preventable diseases for that matter) are not commercially available, a number of groups have validated multiplex bead-based immunoassays for simultaneously detection of antibodies against these three infectious diseases. However it is difficult to compare the assays described in the different studies, as various investigators used different materials, reagents and reference sera and reports varied considerably in the amount of methodological details provided. In our recent study, we developed a Luminex xMAP based assay using for the first time magnetic beads and commercially available purified antigens and commercial ELISA kits for assay comparison, parameters which we think are important for future standardisation among different laboratories.

Optimising a 6-plex tetanus-diphtheria-pertussis fluorescent bead-based immunoassay

“…Three commonly used buffers were compared for potential matrix effects: 1) PBS-T (0.05%), 2) PBS-BSA (1%) and 3) PBS-T (0.05%) - BSA (1%) [3] . Two QC sera (donor serum, 1:1000) and a blank control were run in duplicate on 3 separate plates, changing only the wash buffer used between plates.…”