[Development of a solid-phase immunoenzyme analysis of gentamicin in human blood serum].
An enzyme immune test system was designed and optimized for quantitative assay of gentamicin in human sera. Immunospecific reagents i.e. gentamicin conjugates with ovalbumin (for sorption on polysterol plates) and with bovine serum albumin (immunogen) were prepared. Gentamicin specific antisera were isolated and tested. Conditions for the antigen sorption on polysterol plates were determined and optimized. Different regimes of the competition reaction were investigated and conditions for the antibiotic assay in human sera were determined. The assay specificity was studied and the stability of the test system was checked. An experimental lot of the reagent set was manufactured at the ZAO NPP Immunotech and the correlation tests with the use of the fluorescence polarization immunoassay were performed. The set is destined for the assay of 40 samples (in duplicate). The method sensitivity is 1 ng/ml of gentamicin. The range of the detectable concentration is 1 to 32 ng/ml of gentamicin in 1000-fold diluted sera. The assay time is not more than 3 hours. The variation coefficient of the results does not exceed 12 per cent. The shelf-life of the set is 0.5 years when stored at a temperature of 2 to 8 degrees C.
Kolosova
A Iu
AIu
Department of Chemistry, M.V. Lomonosov Moscow State University.
Blintsov
A N
AN
Samsonova
Zh V
ZhV
Egorov
A M
AM
D000900:Anti-Bacterial Agents / Q000097:blood* / Q000737:chemistry; D002138:Calibration; D004355:Drug Stability; D005839:Gentamicins / Q000097:blood* / Q000737:chemistry; D006801:Humans; D007124:Immunoenzyme Techniques; D016014:Linear Models; D010047:Ovalbumin / Q000737:chemistry; D012680:Sensitivity and Specificity
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D010047:Ovalbumin