Comparison of Five Commercial Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to Bordetella pertussis

Abstract

Measuring antibodies to Bordetella pertussis antigens is mostly done by enzyme-linked immunosorbent assays (ELISAs). We compared the performance of five commercially available ELISA kits with the help of 65 serum specimens which were repetitively tested for evaluation of the kits. The specimens contained 20 paired serum samples from patients with clinical pertussis, 15 samples were from children vaccinated with a diphtheriatetanus-acellular pertussis vaccine, seven specimens were taken from an interlaboratory comparison of ELISAs, and there were three reference preparations from the Food and Drug Administration's (FDA's) Laboratory of Pertussis and from our laboratory. Reference values were obtained from the FDA or from results obtained with an in-house ELISA. Commercial ELISAs were compared with respect to their reproducibility and variability, their ability to detect significant titer rises in paired serum samples, their ability to detect an immune response after vaccination, and the comparability of semiquantitative and quantitative results. Reproducibility was generally good (>89%), intra-assay variation ranged from 2.4 to 28.7%, and indeterminate results were recorded in up to 18.5% of all specimens. Most kits correctly identified the antibody response to an acellular pertussis vaccine. None of the commercial kits identified all cases of pertussis correctly, and the sensitivity ranged between 60 and 95%. All five commercial ELISAs showed great discrepancies when comparing semiquantitative results and contained obviously different antigen preparations. Our data suggest that the five commercial ELISAs tested here need further improvement and standardization.

Laboratory Diagnosis of Pertussis

“…While in-house IgG-Ptx ELISAs accurately distinguished WHO reference preparations and gave results comparable to the expected values, commercial kits using mixtures of antigens did not appear to be able to give results that correlated with those obtained with the WHO reference preparations (223). Comparison of the performances of five commercial ELISAs for detection of antibodies to B. pertussis (two using a mixture of Ptx and FHA and three using an unknown coating antigen) with in-house ELISAs measuring IgG and IgA antibodies to Ptx and FHA according to the CBER protocol resulted in the conclusion that these ELISAs needed further improvement and standardization (260). In a study of convalescentphase sera from 41 patients with culture-or PCR-confirmed pertussis and from 65 control patients with other diseases, the discriminative powers of the IgG-Ptx components of three commercial ELISAs and of the in-house IgG-ELISA were practically identical (AUC of ROC curves, 0.92 to 0.93) (261).…”

Antik�rper gegen impfpr�ventable Erkrankungen bei Schwangeren und deren Neugeborenen

“…Obwohl die ELISA-Technik bislang in zahlreichen Studien zur Bestimmung von Antikörpern gegen B. pertussis eingesetzt wurde,sind bis heute keine standardisierten Testmethoden verfügbar. Die Validierung des eingesetzten Tests wurde durch das Institut für Hygiene und Labormedizin,Klinikum Krefeld, vorgenommen [7].…”

Comparison of Western Immunobloting to an Enzyme-Linked Immunosorbent Assay for the Determination of Anti-Bordetella pertussis Antibodies

“…The different ELISAs contained different antigen preparations. It was concluded from this study that the five tested commercial ELISAs needed further improvement and standardization (17).…”

“…infection (27). The detection of IgA antibodies against PT and FHA suggests natural infection not immunization, since IgA is rarely if at all detected following immunization (17,21). However, the detection of IgG to PT and FHA is more sensitive than that of IgA, since not all individuals mount a detectable IgA response following infection (22).…”