METHOD AND A KIT FOR AN IMMUNOENZYMOMETRIC ASSAY OF THE QUANTITATIVE CONTENT OF ANTITOXIC ANTI-DIPHTHERIA IGG ANTIBODIES IN HUMAN SERUM Russian patent published in 2021 - IPC G01N33/53 G01N33/48 

FIELD: immunology.

SUBSTANCE: invention relates to immunology. To detect antitoxic IgG antibodies in quantitative terms (mcg/ml), a standard calibration sample containing 80 mcg/ml of anti-diphtheria antitoxic IgGs is put into the wells of a microplate with an occluded antigen (10 mcg/ml). The test samples of human blood serum are incubated. Then a conjugate of the horseradish peroxidase enzyme and rabbit antibodies against human IgGs are put into the wells at a dilution of 1: 16000. Incubation is carried out at a temperature of 37 ° C for 30 minutes in a thermal shaker with a frequency of 700 rpm. The contents of the wells are poured and washed four times with a phosphate buffered saline with Tween-20. After that a tetramethylbenzidine solution is added and incubated. A stop reagent is added to all wells of the plate and the formed immune complexes are detected at a wavelength of 450 nm. Calculation of the concentration of the determined antitoxic IgGs is carried out using a standard calibration sample with a known concentration of antitoxic IgGs taking into account the dilution of the samples. The kit for the determination of antitoxic anti-diphtheria antibodies of a person contains: a flat-bottomed microplate with the occluded antigen (10 mcg/ml), a standard calibration sample containing anti-diphtheria antitoxic IgGs (80 mcg/ml), a conjugate of the horseradish peroxidase enzyme with rabbit antibodies against human IgGs, a concentrate of the phosphate buffered saline with the detergent Tween-20, a tetramethylbenzidine solution, a stop reagent, and instructions.

EFFECT: invention increases specificity, sensitivity and reproducibility of results. It also reduces analysis time.

2 cl, 2 tbl, 3 ex, 2 dwg