Comparison of electrochemiluminescence assay and ELISA for the detection of Clostridium botulinum type B neurotoxin.
We compared the ELISA and electrochemiluminescence (ECL) immunoassay technologies for the detection of botulinum type B neurotoxin (BotNT B), which requires highly sensitive techniques due to its potent biological activity. BotNT B complexes are the naturally secreted form of the toxin, approximately a third of which consists of the neurotoxin itself; they were aliquoted and frozen for this study. Results of both techniques were interpreted with the same standard statistical tests (ANOVA and Tukey). We first compared two commercial assays for BotNT B: the detection limit of the colorimetric ELISA was 1.56 ng/ml BotNT B complexes versus 0.39-0.78 ng/ml in the ECL test. We then used the same monoclonal antibody and the same polyclonal antibody, respectively purified by protein A and protein G chromatography, to optimize an in-house ELISA test and an in-house ECL test, making it possible to directly compare the two technologies without interference due to the properties of the antibodies used in the two tests. The colorimetric in-house ELISA had a detection threshold of 3.12 ng/ml versus the in-house ECL test whose detection threshold was 0.78-1.56 ng/ml. Thus, in both cases, the ECL assay was two to four times more sensitive than the colorimetric ELISA. The ECL assay was also more rapid (2.5 h for the in-house ECL versus 5 h for in-house ELISA with precoated wells). Overall, these elements can be used to compare the qualities of the two technologies, at least for the detection of protein antigens such as toxins.
Immunobiologie, Département de biologie des agents transmissibles, Centre de recherche du, Service de santé des armées, 24 avenue des maquis du Grésivaudan, PO Box 38702, La Tronche, France.
V
V
Guglielmo-Viret
O
O
Attrée
V
V
Blanco-Gros
P
P
Thullier
D047014:Clostridium botulinum type B / Q000737:chemistry* / Q000276:immunology*; D004563:Electrochemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D009498:Neurotoxins / Q000032:analysis* / Q000276:immunology*
D009498:Neurotoxins