METHOD FOR PRODUCING STANDARD SERUM PANEL FOR CONTROLLING TEST SYSTEM AND IMMUNOBLOT QUALITY
FIELD: medicine. SUBSTANCE: method involves selecting immunospecific sera containing antibodies to main viral antigens and having the antibodies in various concentrations, and also selecting native donor sera possessing zero titer. Serum selection for producing panels is carried out according to res...
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Zusammenfassung: | FIELD: medicine. SUBSTANCE: method involves selecting immunospecific sera containing antibodies to main viral antigens and having the antibodies in various concentrations, and also selecting native donor sera possessing zero titer. Serum selection for producing panels is carried out according to results of examination by applying polymerase color reaction method for detecting infectious material presence and titering method in enzymoimmune assay. Sera are selected as zero titer sera when the result of polymerase color reaction method is negative and that of enzymoimmune assay shows optical density value to be less than the threshold one. The selected sera are diluted with diluting agent having no antibodies to the virus under study to obtain various antibody concentrations. Specific sera are diluted with nonimmune serum. Stabilizing agent is introduced into all panel sera. Specific sera samples are certified according to definite antibody types availability in them by their testing with enzymoimmune assay with separate antigens of the virus under study or in immunoblot. Minimum number of sera are selected as standard ones so that complete antibody set is provided in the standard serum panel corresponding to main antigens of the virus under study. To build a standard panel, 25-30% of zero titer are selected and not less than 50% of sera with titers from 1:100 to 1:200 and not more than 20-25% of sera with 1: 400 titer, provided the total number of sera is not less than 24. EFFECT: provided accuracy of serologic analysis; high quality of test systems and immunoblots. 6 cl
Изобретение относится к биотехнологии и медицине и может быть использовано в технологии приготовления панели сывороток, содержащих антитела к тестируемому вирусу, для контроля чувствительности и воспроизводимости тест-систем и иммуноблотов. Способ получения стандартной панели сывороток для контроля качества тест-систем и иммуноблотов включает отбор иммуноспецифических сывороток, содержащих антитела к основным антигенам вируса и имеющих разную концентрацию этих антител, а также отбор нативных донорских сывороток с нулевым титром. Отбор сывороток для панелей производится по результатам их исследования методом полимеразной цепной реакции ПЦР на наличие инфекционного материала и титрования в иммуноферментом анализе (ИФА). В качестве иммуноспецифических сывороток выбирают сыворотки, которые в процессе титрования соответствуют закономерному уменьшению величины оптической плотности (ОП) в ИФА и ве |
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