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ПРИМЕНЕНИЕ MALDI МАСС-СПЕКТРОМЕТРИИ В ДИАГНОСТИКЕ ОСОБО ОПАСНЫХ ИНФЕКЦИОННЫХ БОЛЕЗНЕЙ: СОВРЕМЕННОЕ СОСТОЯНИЕ И ПЕРСПЕКТИВЫ ФКУЗ «Российский научно-исследовательский противочумный институт «Микроб», Саратов, Российская ФедерацияМасс-спектрометрия является современным физико-химическим методом анализа, позволяющим проводить качественный и количественный анализ состава вещества, основанный на предварительной ионизации входящих в его состав атомов или молекул. Одним из новых методов ионизации, благодаря которому масс-спектро метрическое исследование макромолекул получило широкое распространение, является разработанная матричноактивированная лазерная десорбция/ионизация (MALDI), представляющая собой импульсное лазерное облучение исследуемого вещества, смешанного с матрицей. В обзоре представлены современные данные о применении метода MALDI масс-спектрометрии для проведения родо-и видоспецифической идентификации микроорганизмов в практике диагностических лабораторий. Рассмотрены преимущества MALDI-TOF идентификации по сравнению с бактериологическими, иммунологическими и молекулярно-генетическими методами исследования. Обозначено место масс-спектрометрии в системе лабораторной диагностики инфекционных болезней, в том числе особо опасных на территории Российской Федерации.Ключевые слова: MALDI масс-спектрометрия, MALDI-TOF, идентификация, патогенные биологические агенты. A.N.Spitsyn, D.V.Utkin, V.E.Kuklev, S.A.Mass spectrometry is a modern physical-chemical analytical method that provides for qualitative and quantitative assessment of the substance composition. It is based on pre-ionization of the atoms and molecules included into it. One of the advanced methods of ionization, due to which mass-spectrometry investigation of macromolecules has become a frequent practice, is matrix-assisted laser desorption/ionization (MALDI). The essence of it is the pulsed laser irradiation of the matter under study, mixed with the matrix. The review discusses current data on MALDI mass-spectrometry application for the performance of species-specific and genus-specific identification of microorganisms at the premises of diagnostic laboratories. Considered are the basic advantages of MALDI-TOF identification as compared to bacteriologic, immunologic, and molecular-genetic methods of assessment. Allocated is the mass-spectrometry position in the system of laboratory diagnostics of infectious diseases, including particularly dangerous ones, in the territory of the Russian Federation.

The possibility of characterizing fluorescent immunoglobulins using spectrophotometric analysis as a testing method is considered. The comparative analysis of optical properties of fluorescent immunoglobulin preparations and their components-immunoglobulins and fluorochrome-is carried out. The obtained results testify that the proposed methodological approach of optical detection of labeled immunoglobulin molecules can be promising for tests on obtaining conjugates used in immunological tests on revealing specific antigens of causative agents of especially dangerous infections.

The possibility of detecting the formation of immune complexes with the use of antibodies labeled with colloidal gold nanoparticles and optical spectroscopy is considered. Changes in the spectral characteristics of sols of gold nanoparticles with diameters of 15, 60, and 90 nm conjugated with immunoglobulins in the presence of especially dangerous infectious agents are recorded and analyzed. The data obtained indicate that the developed methodological approach to optical detection of immune complexes may be promising for creating immunochemical methods of identification of infectious agents.

The atomic power microscopy (APM) is used to study the cell surface structure of particularly dangerous infectious diseases agents and to carry out the morphometric analysis. APM shows similar results with scanning electron microscopy. However, its application makes it possible to avoid time-consuming and labour-intensive procedures of samples preparing for testing by fixation, dehydration and sputtering of conducting layer. Methodological approach has been elaborated for preparing and analysis of samples of agents of particularly dangerous infectious diseases by means of APM. This approach includes a selection of optimal substrate, mode of disinfection and scanning of samples.