METHOD FOR ESTIMATION OF EFFECIENCY OF VACCINATION AGAINST PERTUSSIS, DIPHTHERIA AND TETANUS

摘要

FIELD: medicine.;SUBSTANCE: method consists in semiquantitative determination of postvaccinal immunity to antigens of pertussis, diphtheria and tetanus. For this, the solid phase, which is a test strip made of a nitrocellulose membrane with a pore diameter of 0.45 mcm and size of 0.5×2 cm, are sensitized by application of diphtheria 3.5 μl at a concentration of 0.1 mg/ml and tetanus 3.5 μl with a concentration of 0.05 mg/ml of anatoxins, whole cell pertussis antigen 3.5 μl, 20 MOE, and 3,5 μl of mouse IgG solution at a concentration of 0.1 mg/ml as a positive control. After test strips drying, incubating with a blocking solution of PBST in the wells of a plate placed on a shaker in a thermostat at a temperature of 37°C for 60 min, the solid phase is washed three times for 5 minutes with PBST, incubated with the test serum samples and control serum samples for 30 minutes at 37°C. The antibodies contacted to the solid phase are washed and "developed" using a diagnosticum, which is a conjugate of carbon nanoparticles with streptococcus G protein diluted at 1/45 for 60 minutes. The result is evaluated using a program of scanned imagesr analysis after test strips scanning in an ordinary plate scanner. With an anti-tetanus and antidiphtheria antibody content of less than 0.1 IU/ml, an inadequate protection is estimated for which immunization is required. At 0.1-1.0 IU/ml - a sufficient level of protection, which requires a booster dose of vaccine, and at values greater than 1.0 IU/ml - a high level of protection, and immunization required in 5 years or more. The presence of the protective titer of anti-pertussis antibodies is evaluated by calculating the threshold value: X=0.128(A-B)+B, where X is the threshold value of optical density, A is the optical density of a strongly positive sample, B is the optical density of a weakly positive sample, and if the obtained value of sample optical density is above the threshold value, it contains a protective level of anti-pertussis antibodies.;EFFECT: application of this method allows to use a test system to quantify the presence of protective postvaccinal immunity to three infections: pertussis, diphtheria and tetanus, simultaneously.;2 tbl, 2 ex