OPTIMIZATION OF THE CONDITIONS FOR THE DEVELOPMENT OF THE RESULTS OF THE DOT-IMMUNOASSAY IN AN AUTONOMOUS KIT FOR THE DETECTION OF ORTHOPOXVIRUSES


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Abstract

The aim of the work is to create a sensitive, fast and easy-to-use immunochemical assay for the detection of orthopoxviruses in the “point of care” format. The objectives of the study included: the fine-tuning of technology of preparation and use of the developer for colloidal gold conjugates in autonomous diagnostic systems based on flat protein matrices, as well as testing of this technology for detecting vaccinia virus. The quality of available reagents (metol, citric acid and silver nitrate) for the manufacture of the developing system was assessed: the most suitable reagents were selected. A technology has been fine-tuned for the preparation of a developer. The developer includes the dry component in the form of tablets (3.5-4 mg) containing metol and citric acid in a ratio of 2:3, and the liquid component - 0.4% silver nitrate solution. To increase the contrast and stability of optical signals, it was proposed to use the processing of the developed protein matrix with an alkaline solution of thiourea. The use of a two-component developer allows the detection of vaccinia virus in non-laboratory conditions with a sensitivity of 5.4 × 104 PFU/ml.

About the authors

A. G Poltavchenko

Federal Budgetary Research Institution - State Research Center of Virology and Biotechnology «Vector»

доктор биол. наук, вед. науч. сотр. отдела разработки и производства средств диагностики вирусных заболеваний ФБУН ГНЦ ВБ «Вектор» Роспотребнадзора 630559, Kol’tsovo, Novosibirsk region, Russia

A. V Ersh

Federal Budgetary Research Institution - State Research Center of Virology and Biotechnology «Vector»

канд. биол. наук, науч. сотр. лаб. иммунохимической диагностики ФБУН ГНЦ ВБ «Вектор» Роспотребнадзора 630559, Kol’tsovo, Novosibirsk region, Russia

Mamedyar Sh. Azaev

Federal Budgetary Research Institution - State Research Center of Virology and Biotechnology «Vector»

Email: azaev@vector.nsc.ru
Dr .Biol., Assoc. Prof., Head of the Department of scientific and methodological training of doctors and biologists to work with pathogens of high-risk viral infections of Federal Budgetary Research Institution State Research Center of Virology and Biotechnology “Vector”, Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, «State Research Center of Virology and Biotechnology “Vector”», 630559, Kol’tsovo, Novosibirsk Region, Russian Federation 630559, Kol’tsovo, Novosibirsk region, Russia

P. V Filatov

Federal Budgetary Research Institution - State Research Center of Virology and Biotechnology «Vector»

канд. биол. наук, науч. сотр. лаб. иммунохимической диагностики ФБУН ГНЦ ВБ «Вектор» Роспотребнадзора 630559, Kol’tsovo, Novosibirsk region, Russia

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