Amperometric bioelectrode for specific human immunoglobulin G determination: Optimization of the method to diagnose American trypanosomiasis
| Autor: | María E, Ribone, María S, Belluzo, Daniela, Pagani, Iván S, Marcipar, Iván S, Macipar, Claudia M, Lagier |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Trypanosoma cruzi
Coefficient of variation Biophysics Antibodies Protozoan Enzyme-Linked Immunosorbent Assay Biosensing Techniques Sensitivity and Specificity Biochemistry Immunoglobulin G Antigen Electrochemistry Animals Humans Chagas Disease Electrodes Molecular Biology chemistry.chemical_classification Chromatography biology Chemistry Reproducibility of Results Substrate (chemistry) Cell Biology Amperometry biology.protein Thiol Antibody Biosensor |
| Zdroj: | Analytical Biochemistry. 350:61-70 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2005.11.033 |
| Popis: | Bioelectrodes to detect immunoglobulin G (IgG) antibodies occurring in sera of patients suffering from American trypanosomiasis were assembled. The device consisted of a gold electrode modified with a thiol sensitized with parasite proteins. The assemblage was accomplished by adsorbing IgG antibodies from confirmed infected patients followed by adsorption of anti-human IgG labeled with a redox enzyme. The appliance was used as a working electrode in a three-electrode cell containing a soluble charge-transfer mediator, also behaving as enzyme cosubstrate. The method is based on the measurement of the catalytic current after addition of the enzyme substrate, occurring when a positive serum is used to build up the biosensor. The discrimination efficiency between positive and negative sera was 100% for the samples studied. A 0.9525 correlation coefficient was obtained for results acquired by using this approach and one commercial diagnostic kit. The reproducibility, evaluated by the percentage coefficient of variation, varied between 7 and 20%. The sensitivity was 12.4 ng mL−1 IgG, which is in the same order as that obtained with the commercial kit. Stability of the device was studied for a 7-day period and the results showed no significant change (p = 0.218). Leishmaniasic sera showed cross-reactivity when total parasite homogenate was used as antigen. |
| Databáze: | OpenAIRE |
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