EXAMINATION AND TESTING OF EXPERIMENTAL-PRODUCTION SERIES OF POLYVALENT BRUCELLOSIS SERUM

At present no commercial polyvalent brucellosis serum is produced in Russia. We developed a technology of polyvalent brucellosis high-effective and specific serum production without adsorption. Experimental-production series are obeyed the technical specification project

Improvement of a test-system for the botulinus toxin screening in dot-immunoanalysis

We constructed a test system for dot-immunoassay (DIA) to accelerate definition and identification of botulinus toxins and also to refuse from application of laboratory animals for routine screening of clinical samples, foodstuff and environments. This system permitted to detect botulinus toxin during approximately 2 h in the tested samples. Sensitivity of this DIA in some cases exceeded the mice biotest. This improved method has minimum reaction to nonspecific exposures from the investigated biological substrata. It is simple to conduct. It is high efficient and expressive, does not require to use expensive equipment and the reactants, special training for the personnel. Lyophilization conditions for the immune reagents used for the test system preparation for botulinus toxin dot-immunoassay were selected. High sensitivity, specificity of the analysis are remained, stability of the preparations (periods of storage) is increased. This method is convenient to use in field conditions at extreme situations, in particular, in mobile autolaboratories for epidemiological survey

Investigation of 5’-norcarbocyclic nucleoside analogues as antiprotozoal and antibacterial agents

Carbocyclic nucleosides have long played a role in antiviral, antiparasitic, and antibacterial therapies. Recent results from our laboratories from two structurally related scaffolds have shown promising activity against both Mycobacterium tuberculosis and several parasitic strains. As a result, a small structure activity relationship study was designed to further probe their activity and potential. Their synthesis and the results of the subsequent biological activity are reported herein

PREPARATION OF MONOSPECIFIC BRUCELLA ANTI-ABORTUS SERUM BASED ON THERMOEXTRACT FROM S-FORM BRUCELLA

Objective of the study is to prepare rabbit monospecific anti-abortus serum to a Brucella S-form thermoextract, as well as to evaluate its activity and specificity. Materials and methods. Utilized was polyvalent hyperimmune serum obtained to Brucella thermoextract. It was adsorbed by a strain of heterologous species Brucella melitensis 16 N. Reference, vaccinal and field Brucella strains were used to estimate the activity and specificity of the serum produced. Results and conclusions. This serum possesses high activity (1:160, 1:320 titer) and specificity, and the results completely coincide with the data specified in the table of differential properties presented in МR 3.1.7.1189-03 “Prophylaxis and laboratory diagnostics of human brucellosis». The outlined method for manufacturing monospecific Brucella anti-abortus serum permits for a considerable decrease in biological hazard for personnel involved in the technological operations

PECULIARITIES OF LABORATORY DIAGNOSTICS OF EXPERIMENTAL BRUCELLOSIS CAUSED BY SAND L-FORMS OF CAUSATIVE AGENT

Comparative evaluation, of bacteriological, serological and PCR diagnostic methods was resulted, at experimental brucellosis in guinea pigs infected, with. S- and. L-forms of the causative agent in different periods of the infection. It was shown that Brucella S-forms were isolated, during six months, L-forms - about one month, by a bacteriological method. Positive PCR results were observed about six months after S- and. L-form infection in animals. Positive slide agglutination, of guinea pig blood sera was revealed, till 12 months by a corpuscular diagnosticum with. S- and. L-forms of Brucella. Complex use of bacteriological, PCR and. serological methods for laboratory diagnostics of atypical brucellosis caused, by Brucella L-form allows to obtain more reliable information, about epizootological-epidemiological situation in non-manifested (non-clinical) infection foci

АНАЛИЗ МУТАЦИЙ МИКОБАКТЕРИЙ ТУБЕРКУЛЕЗА, ОПРЕДЕЛЯЮЩИХ ИХ ЛЕКАРСТВЕННУЮ УСТОЙЧИВОСТЬ У БОЛЬНЫХ С НЕЛЕЧЕННЫМ ТУБЕРКУЛЕЗОМ ЛЕГКИХ ПРИ РАЗНОМ ВИЧ-СТАТУСЕ В СВЕРДЛОВСКОЙ ОБЛАСТИ

Goal of the study: to identify profile of mutations of tuberculous mycobacteria responsible for resistance to anti-tuberculosis drugs in HIV positive and HIV negative tuberculosis patients without prior history of treatment.Materials and methods. 165 strains of tuberculous mycobacteria from HIV positive patients and 166 strains of tuberculous mycobacteria from HIV negative patients were studied in Sverdlovsk Region (TB Dispensary, Yekaterinburg). Mutations in genes were identified using microchips of TB-BIOCHIP® and TB-BIOCHIP®-2 in compliance with the manufacturer's guidelines (OOO Biochip-IMB, Moscow).Results. It was observed that 85/165 (51.52%) strains isolated from HIV positive tuberculosis patients and 58/166 (34.94%) strains isolated from tuberculosis patients not associated with HIV possessed MDR genotype (p < 0.01). The majority of MDR strains had mutations in the 531th codon of rpoB (Ser→Leu) and 315th codon of katG (Ser→Thr) (64/85, 75.29% and 38/58, 65.52% respective the groups), resulting in the high level of resistance to rifampicin and isoniazid. Each group also had approximately equal ratio (11/165, 6.67% and 12/166, 7.23% respective the groups) of strains with genomic mutations defining the resistance to isoniazid, rifampicin and fluoruquinolones. No confident difference was found in mutation patterns of genome of tuberculous mycobacteria isolated from HIV positive and HIV negative tuberculosis patients. Цель исследования: определить спектр мутаций МБТ, ответственных за устойчивость к противотуберкулезным препаратам, у больных нелеченным туберкулезом, имеющим ВИЧ-позитивный или ВИЧ-негативный статус.Материалы и методы. Исследовано 165 штаммов МБТ от ВИЧ-позитивных и 166 штаммов МБТ от ВИЧ-негативных больных из Свердловской области (ГБУЗ СО «Противотуберкулезный диспансер», г. Екатеринбург). Определение мутаций в генах проводили на микрочипах «ТБ-БИОЧИП®» и «ТБ-БИОЧИП®-2» согласно инструкции изготовителя (ООО «Биочип-ИМБ», Москва).Результаты. Показано, что 85/165 (51,52%) штаммов, выделенных от больных туберкулезом с ВИЧ-инфекцией, и 58/166 (34,94%) штаммов от больных туберкулезом, не ассоциированным с ВИЧ-инфекцией, обладали МЛУ-генотипом (p < 0,01). Подавляющее большинство МЛУ-штаммов имели мутации в 531-м кодоне rpoB (Ser→Leu) и 315-м кодоне katG (Ser→Thr) (64/85, 75,29% и 38/58, 65,52% соответственно по группам), приводящие к высокому уровню резистентности к рифампицину и изониазиду. Кроме того, в каждой группе приблизительно в равном соотношении (11/165, 6,67% и 12/166, 7,23% соответственно по группам) встречались штаммы, несущие в геноме мутации, определяющие устойчивость к изониазиду, рифампицину и фторхинолонам. Достоверных отличий по спектру мутаций в геноме МБТ, выделенных от ВИЧ-позитивных и ВИЧ-негативных больных туберкулезом, не показано

Сравнительная молекулярно-генетическая характеристика культур Mycobacterium tuberculosis, выделенных в Европейской части Российской Федерации в 1998–2003 гг. и 2016–2021 гг.

The objective: to evaluate changes in detection frequency of Mycobacterium tuberculosis of the main phylogenetic lines with various genetic determinants of resistance to rifampicin, isoniazid and fluoroquinolones in 1998–2003 and 2016–2021.Subjects and Methods. 965 mycobacterial cultures were studied, those cultures were isolated from pulmonary tuberculosis patients in 1998–2003 and 2016–2021. The spoligotypes of isolated Mycobacterium tuberculosis cultures and presence of gene mutations associated with resistance to isoniazid, rifampicin, and fluoroquinolones were determined.Results. In 2016–2021 versus 1998–2003, the incidence of Beijing subline increased from 50,72% to 64,60%. Within Euro-American line, the detection frequency of subline LAM9 decreased (from 40,00% to 14,81%), while the detection frequency of sublineT1 increased (from 27,11% to 41,36%). Mycobacterium tuberculosis with MDR genotype rpoB531_Ser->Leu + katG315_Ser-> Thr [1] was selected. Mycobacterium tuberculosis with genotypic resistance to fluoroquinolones was detected only in cultures of 2016–2021, and Mycobacterium tuberculosis with preXDR genotype was significantly more common in Beijing subline compared to Euro-American line (21,67% vs. 3,73%, p <0,05).Цель исследования: оценить динамику частоты выявления МБТ основных филогенетических линий с различными генетическими детерминантами устойчивости к рифампицину, изониазиду и фторхинолонам в периоды 1998–2003 гг. и 2016–2021 гг.Материалы и методы. Исследовано 965 культур микобактерий, выделенных от больных туберкулезом легких в 1998–2003 гг. и 2016–2021 гг. Определены сполиготипы выделенных культур МБТ и наличие мутаций в генах, ассоциированных с устойчивостью к изониазиду, рифампицину и фторхинолонам.Результаты. В 2016–2021 гг. по сравнению с 1998–2003 гг. повысилась частота встречаемости МБТ Пекинской сублинии с 50,72% до 64,60%. В рамках Евро-Американской линии снизилась частота выявления МБТ сублинии LAM9 (с 40,00% до 14,81%) с повышением частоты выявления МБТ сублинии T1 (с 27,11% до 41,36%). Произошел отбор МБТ с МЛУ-генотипом rpoB531_Ser->Leu + katG315_Ser->Thr [1]. МБТ с генотипической устойчивостью к фторхинолонам были выявлены только в культурах 2016–2021 гг., причем МБТ с преШЛУ-генотипом достоверно чаще встречались у МБТ Пекинской сублинии по сравнению с МБТ Евро-Американской линии (21,67% против 3,73%, p<0,05)

Construction of nutrient media for L-form brucella

Dry nutrient media on the basis of hepatic infusion and Siberian roach hydrolyzate are constructed. Experimental data demonstrate that these media possess high sensitivity, completely inhibit growth of S-form Brucella, do not require рН adjustment, filtration and autoclaving. They can be used for isolation, cultivation and accumulation of L-form Brucella for bacteriological diagnostics of brucellosis. As these media are accessible for transportation, they are applicable for stationary and field conditions

Microbiological diagnosis of coincident mycobacterial infection in cystic Fibrosis

Diseases caused by mycobacteria, leprosy and tuberculosis, are widely known for many centuries. However, interest in mycobacteriosis, as an infectious disease of human organs caused by other mycobacteria, was only apparent from the late 50s of the 20th century. Over the years of observation, the number of cases of mycobacteriosis in various regions of the world has increased significantly, including in patients with cystic fibrosis. A key sign of comorbidity in cystic fibrosis is a chronic microbial infection that determines the severity of the course and the prognosis of the disease. In order to identify and identify mycobacteria isolated from patients with cystic fibrosis from 2011 to 2018, sputum was examined from 160 patients with cystic fibrosis and suspected mycobacterial infection. Mycobacteria were secreted in 17 (10.6%) patients with cystic fibrosis, of which non-tuberculous mycobacteria predominated. Thirteen patients were infected with M. abscessus (76.4%), two with M.avium (11.7%), one with M.xenopi (5.8%), one with M. tuberculosis (5.8%). Determination of drug sensitivity of mycobacteria showed that the culture of M.tuberculosis was sensitive to all antituberculosis drugs, and strains of nontuberculous mycobacteria had a different spectrum of drug susceptibility to drugs used for the therapy of mycobacteriosis. Identification of infectious agents will help ensure the timely initiation of adequate treatment, proper monitoring and prevention of the spread of non-tuberculosis mycobacteria among patients with cystic fibrosis.Заболевания, вызываемые микобактериями, лепра и туберкулез, широко известны на протяжении многих столетий. Однако интерес к микобактериозу, как инфекционному поражению органов человека, вызываемому другими микобактериями, проявился только с конца 50-х годов ХХ века. За годы наблюдений число случаев заболевания микобактериозом в различных регионах мира значительно увеличилось, в том числе и у больных муковисцидозом. ключевой признак коморбидности при муковисцидозе - хроническая микробная инфекция, которая определяет тяжесть течения и прогноз заболевания. С целью выявления и идентификация микобактерий, выделенных от больных муковисцидозом за период с 2011 по 2018 год, была исследована мокрота от 160 пациентов с муковисцидозом и подозрением на микобактериальную инфекцию. У 17 (10,6%) пациентов с муковисцидозом были выделены микобактерии, из них преобладали нетуберкулезные микобактерии. Тринадцать пациентов были инфицированы M.abscessus(76,4%), два - M.avium (11,7%), один - M.xenopi (5,8%), один - M.tuberculosis (5,8%). определение лекарственной чувствительности микобактерий показало, что, культура M.tuberculosis была чувствительна ко всем противотуберкулезным препаратам, а штаммы нетуберкулезных микобактерий имели различный спектр лекарственной чувствительности к препаратам, применяемым для терапии микобактериозов. Идентификация возбудителей инфекции будет способствовать обеспечению своевременного начала адекватного лечения, надлежащего мониторинга и профилактики распространения нетуберкулезных микобактерий среди больных кистозным фиброзом (муковисцидозом)