RU94029693A - Method for estimating pertussis preparation toxicity - Google Patents

Method for estimating pertussis preparation toxicity

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Publication number
RU94029693A
RU94029693A RU94029693/14A RU94029693A RU94029693A RU 94029693 A RU94029693 A RU 94029693A RU 94029693/14 A RU94029693/14 A RU 94029693/14A RU 94029693 A RU94029693 A RU 94029693A RU 94029693 A RU94029693 A RU 94029693A
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RU
Russia
Prior art keywords
pertussis
toxicity
preparations
cells
toxic
Prior art date
Application number
RU94029693/14A
Other languages
Russian (ru)
Other versions
RU2092844C1 (en
Inventor
Е.П. Москаленко
С.Ю. Тюкавкина
Original Assignee
Е.П. Москаленко
С.Ю. Тюкавкина
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Е.П. Москаленко, С.Ю. Тюкавкина filed Critical Е.П. Москаленко
Priority to RU94029693A priority Critical patent/RU2092844C1/en
Publication of RU94029693A publication Critical patent/RU94029693A/en
Application granted granted Critical
Publication of RU2092844C1 publication Critical patent/RU2092844C1/en

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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

FIELD: medicine. SUBSTANCE: method involves adding pertussis preparations at various doses (0.1-10.0 IU for corpuscular preparations and 1.0-100.0 mcg with respect to protein for separate antigens of pertussis microbe or their complexes) to 1.0 ml of monocyte-like human cell line suspension I 937 (4 x 10cells/ml) and in 5-7 h after their being cultivated together in incubator at 37 C on cover glasses in penicillin bottles containing 1 ml of medium 199 having 20% of cattle serum, the cover glasses are removed from the penicillin bottles, washed in Hanks solution, fixed in Nikiforov mixture, stained after Romanovsky-Gimza, and 300 cells are examined by means of light microscope to determine injured cells percentage. Pertussis preparation toxicity is evaluated based on injured cells percentage increase in comparison with control sample as light toxicity when 10-20%, ordinary toxicity when 20-40%, high toxicity level when more than 40% increase is observed. EFFECT: enhanced accuracy of the method; high sensitivity.

Claims (1)

Изобретение относится к медицинской микробиологии и может быть использовано для проверки токсических свойств коклюшных вакцинных препаратов, выпускаемых в серийное производство. Цель изобретения - повышение информативности и достоверности, а также ускорение и упрощение способа оценки токсичности коклюшных препаратов. Способ осуществляется следующим образом. К 1,0 мл взвеси перевиваемой моноцитоподобной линии клеток человека И 937 (4•106кл/мл) добавляют коклюшные препараты в разных дозировках (0,1 - 10,0 МОЕ для корпускулярных препаратов и 1,0 - 100,0 мкг по белку для отдельных антигенов коклюшного микроба или их комплексов) и через 5-7 ч совместного культивирования в термостате при 37°С на покровных стеклах в пенициллиновых флаконах с 1 мл среды 199, содержащей 20% сыворотки крупного рогатого скота, покровные стекла извлекают из пенициллиновых флаконов, промывают в растворе Хэнкса, фиксируют в смеси Никифорова, окрашивают по Романовскому-Гимзе, просматривают 300 клеток в световом микроскопе, определяют процент поврежденных клеток и по увеличению процента поврежденных клеток по сравнению с контролем оценивают токсичность коклюшных препаратов: 10-20% - слабо токсичен; 20-40% - токсичен; более 40% - очень токсичен. Предлагаемый cпособ более информативен, экономичен, сокращает сроки исследования токсичности коклюшных препаратов в 20 раз, не требует лабораторных животных, отличается высокой чувствительностью и хорошей воспроизводимостью.The invention relates to medical microbiology and can be used to test the toxic properties of pertussis vaccine preparations produced in mass production. The purpose of the invention is to increase the information content and reliability, as well as accelerating and simplifying the method for assessing the toxicity of pertussis drugs. The method is as follows. To 1.0 ml of the suspension of the transplanted monocyte-like human cell line And 937 (4 • 10 6 cells / ml), pertussis preparations are added in different dosages (0.1 - 10.0 MOU for corpuscular preparations and 1.0 - 100.0 μg per protein for individual pertussis microbial antigens or their complexes) and after 5-7 hours of co-cultivation in an incubator at 37 ° C on coverslips in penicillin vials with 1 ml of medium 199 containing 20% cattle serum, coverslips are removed from penicillin vials washed in Hanks solution, fixed in a mixture of Nikiforov , Stained according to Romanovsky-Gimza, scan cells 300 in the light microscope, the percentage of damaged cells and by an increase in percent of injured cells compared with the control evaluate the toxicity of pertussis preparations: 10-20% - slightly toxic; 20-40% - toxic; more than 40% - very toxic. The proposed method is more informative, economical, reduces the time of the study of the toxicity of pertussis drugs by 20 times, does not require laboratory animals, is highly sensitive and has good reproducibility.
RU94029693A 1994-08-05 1994-08-05 Method of evaluation of vaccine whooping cough preparation toxicity RU2092844C1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
RU94029693A RU2092844C1 (en) 1994-08-05 1994-08-05 Method of evaluation of vaccine whooping cough preparation toxicity

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
RU94029693A RU2092844C1 (en) 1994-08-05 1994-08-05 Method of evaluation of vaccine whooping cough preparation toxicity

Publications (2)

Publication Number Publication Date
RU94029693A true RU94029693A (en) 1996-06-20
RU2092844C1 RU2092844C1 (en) 1997-10-10

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Family Applications (1)

Application Number Title Priority Date Filing Date
RU94029693A RU2092844C1 (en) 1994-08-05 1994-08-05 Method of evaluation of vaccine whooping cough preparation toxicity

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RU (1) RU2092844C1 (en)

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Publication number Publication date
RU2092844C1 (en) 1997-10-10

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