Respiratory Function of Mitochondria in Patients with Coronary Heart Disease with Implantable Cardioverter-Defibrillator.

We studied the respiratory activity of mitochondria in peripheral blood leukocytes from 36 patients with coronary heart disease (CHD) and a history of ventricular tachyarrhythmias required cardioverter-defibrillator implantation. The measurements were carried out in incubation buffers with different oxidation substrates (succinate and pyruvate-malate mixture). In pyruvate-malate incubation buffer, oxygen consumption rate and respiratory control coefficients in patients with triggered device did not differ significantly from those in patients without cardioverter-defibrillator triggering. At the same time, respiratory control coefficients were below the reference values. In succinate buffer, values of mitochondrial parameters were significantly lower in patients with triggered devices. Our findings indicate that mitochondria of patients with non-triggered cardioverters-defibrillators have better functional and metabolic plasticity. It was concluded that activity of respiratory processes in mitochondria could be an indicator that should be taken into the account when assessing the risk of developing ventricular tachyarrhythmias.

Relationship of Inotropic Effects of Stimulation of ß1- and ß2-Adrenergic Receptors of Isolated Myocardial Fragments with Echocardiography Parameters in Coronary Heart Disease.

We studied the relationship of inotropic responses of the isolated myocardium to stimulation of ß1-and ß2-adrenergic receptors (ß-AR) with echocardiography parameters in 28 patients with coronary heart disease (CHD). Myocardial fragments (trabeculae of the right atrial appendage) were obtained during coronary artery bypass surgery. The inotropic response of the trabeculae was assessed in an isometric mode. Stimulation of ß1-and ß2-AR with agonists was performed against the background of preliminary α-AR blockade. In case of preserved ejection fraction, significant inotropic response of the trabeculae (135 (112; 154)% from the initial contraction amplitude) was observed after ß1-AR stimulation, while in reduced ejection fraction, its significant increase was observed after ß1-AR stimulation (126 (112; 170)% from the initial contraction amplitude). In patients with preserved and reduced ejection fraction, the correlations between the inotropic responses of the trabeculae to ß1-and ß2-AR stimulation and echocardiography parameters were different. The revealed differences reflect the degree of cardiac remodeling under condition of the studied pathology.

[Methylation of Regulatory Regions of DNA Repair Genes in Carotid Atherosclerosis].

The status of DNA methylation in the human genome changes during the pathogenesis of common diseases and acts as a predictor of life expectancy. Therefore, it is of interest to investigate the methylation level of regulatory regions of genes responsible for general biological processes that are potentially significant for the development of age-associated diseases. Among them there are genes encoding proteins of DNA repair system, which are characterized by pleiotropic effects. Here, results of the targeted methylation analysis of two regions of the human genome (the promoter of the MLH1 gene and the enhancer near the ATM gene) in different tissues of patients with carotid atherosclerosis are present. Analysis of the methylation profiles of studied genes in various tissues of the same individuals demonstrated marked differences between leukocytes and tissues of the vascular wall. Differences in methylation levels between normal and atherosclerotic tissues of the carotid arteries were revealed only for two studied CpG sites (chr11:108089866 and chr11:108090020, GRCh37/hg19 assembly) in the ATM gene. Based on this, we can assume the involvement of ATM in the development of atherosclerosis. "Overload" of the studied regions with transcription factor binding sites (according to ReMapp2022 data) indicate that the tissue-specific nature of methylation of the regulatory regions of the MLH1 and ATM may be associated with expression levels of these genes in a particular tissue. It has been shown that inter-individual differences in the methylation levels of CpG sites are associated with sufficiently distant nucleotide substitutions.

[Five-year dynamics adrenergic reactivity of erythrocytes after radio-frequency sympathic denervation of renal arteries in patients with resistant arterial hypertension].

AIM: To study the initial state of adrenergic reactivity and the five-year dynamics of the beta-adrenergic reactivity index of erythrocyte membranes and the manifestation of the antihypertensive effect of the procedure for radiofrequency destruction of sympathetic structures of the renal artery in patients with resistant arterial hypertension. SUBJECTS AND METHODS: The analysis included 42 patients with resistant arterial hypertension (RH). The renal denervation (RD) procedure of the kidneys was performed by endovascular bilateral transcatheter radiofrequency ablation of the renal arteries. The study of 24-hour blood pressure monitoring (BPM) and the determination of ß-adrenoreactivity of erythrocytes (ß-ARM) by changes in the osmoresistance of erythrocyte membranes were performed initially, 1 week, 6 months, 1, 2, 3 and 5 years after RD. Patients retrospectively, at a follow-up period of 6 months after RD, were divided into responders (decrease in blood pressure by 10 or more mm Hg) and non-responders (decrease in blood pressure less than 10 mm Hg). RESULTS: 6 months after the RD, the number of responders was 28 people (66.7%), after 5 years - 31 people (73.8%). At the time of inclusion in the study, the median ß-ARM in the group of non-responders was not significantly higher than in the group of responders. After 6 months after the RD procedure, the ß-ARM indicator in the non-responder group was significantly lower than in the responder group (p = 0.043). With further follow-up in the group of responders, an increase in the median ß-ARM was noted, which reached significant differences relative to the baseline values in the group at follow-up periods of 1 year (p = 0.036) and 5 years (p = 0.004) after RD. The change in the ß-ARM indicator in the non-responder group was wavy in nature, the changes did not reach the significance criteria. CONCLUSION: Renal denervation in 73.8% of cases is accompanied by a stable antihypertensive response for 5 years of observation and an increase in ß-ARM, which may indicate the implementation of compensatory mechanisms in conditions of increasing activity of the sympathoadrenal system in response to a decrease in blood pressure.

Effectiveness of using liquid transport media in bacteriological diagnostics of diphtheria infection.

The results of evaluating the effectiveness of the use of liquid transport media at the preanalytical stage of bacteriological diagnosis of diphtheria infection are presented. A typical toxigenic strain of C. diphtheriae biovar gravis № 665 was used. The experiments were carried out using a laboratory-prepared medium based on GRM-broth (State research center for applied biotechnology and microbiology, Obolensk), a transport system with a fleecy probe swab (DELTALAB) and a transport system ∑-Transwab ® with a polyurethane Sigma-swab (Medical Wire & Equipment Co. (Bath) Ltd.). The tampons were pooled with a 24-hour bacterial culture of C. diphtheriae, then immediately seeded on Tellurite-containing blood agar. Storage conditions were simulated for 6-24 hours: at room conditions +(20-25)° C, in the refrigerator +(4-8)° C, in a thermostat +(37±1)° C. Storage of C. diphtheriae was most optimal on two liquid transport systems in a refrigerator +(4-8)° C for 6 and 24 hours; in room conditions +(20-25)° C - there was a decrease in seeding after 6 hours and loss of pathological material after 24 hours, more pronounced on a fleecy probe swab; under thermostat conditions +(37±1)° C on both transport systems, a decrease in seeding was noted after 6 hours and a complete loss of pathological material after 24 hours. The results obtained demonstrated the efficiency of using the Amies liquid transport medium and justify the need to develop a domestic analogue of the transport system based on the Amies liquid medium for the bacteriological diagnosis of diphtheria infection.

Effect of different types of dry swabs and their storage conditions on the inoculation of Corynebacterium diphtheriae.

The results of evaluating the effectiveness of C. diphtheriae inoculation using different types of dry swabs in studies simulating various conditions of its storage at the preanalytical stage of a laboratory study for diphtheria are presented. A typical toxigenic strain of C. diphtheriae biovar gravis No. 665 was used. A commercial dry, sterile cotton swab probe (Ningbo Greetmed Medical Instruments Co., LTD, China), a commercial dry, sterile swab probe (plastic and viscose) (COPAN, Italy), tufters with a fluffy probe-tampon on a polystyrene applicator, standard (DELTALAB, SL, Spain). The tampons were pooled with a 24-hour bacterial culture of C. diphtheriae, then immediately seeded on Tellurite-containing blood agar and Corynebacagar. Storage conditions were simulated for 3 hours: at room conditions +(20-25)°C, in the refrigerator +(4-8)°C, in a thermostat +(37±1)°C. Optimal storage of C. diphtheriae on all three types of dry swabs at + (4-8) ° C; at +(20-25)° C - growth is observed when seeding from a cotton swab; in a swab with a fleecy probe-tampon, a decrease in the inoculation of C. diphtheriae was noted; when using a viscose swab - a significant loss of C. diphtheriae. At +(37±1)°C, a significant decrease in the inoculation of C. diphtheriae on all three types of tampons was noted, up to the absence of growth when using a viscose tampon. To exclude the loss of C. diphtheriae, it is necessary to observe the conditions for taking and storing biological material at the preanalytical stage of a laboratory study, which will improve the quality of laboratory microbiological studies for diphtheria infection.

Age Peculiarities of Respiratory Activity and Membrane Microviscosity of Mitochondria from Rat Cardiomyocytes.

We studied respiratory activity and microviscosity of mitochondrial membranes from the left ventricular myocardium of 2-month-old (n=8) and 15-month-old (n=8) rats. The respiratory control (RC) during substrate phosphorylation was calculated as the ratio of oxygen absorption rates in the presence of ADP and after its utilization. In 2-month-old rats, RC was 4.66 (4.56; 4.71); in 15-month-old animals, it was significantly (р<0.05) lower: 3.57 (3.50; 3.62). Pyrene probe eximerization indices for regions of protein-lipid and lipid-lipid interactions in mitochondria of 2-month-old animals were significantly lower than in the group of 15-month-old rats, which indicated reduced microviscosity of the lipid environment of proteins. Thus, the decrease in RC of mitochondria from the left ventricle of 15-month-old animals and the increase in the microviscosity coefficients of their membranes indicate age-related changes in the structural and functional activity of mitochondria.

Erythrocyte membranes beta-adrenoreactivity changes after renal denervation in patients with resistant hypertension, relationship with antihypertensive and cardioprotective intervention efficacy.

Aim      To study the functional condition of sympathoadrenal system as evaluated by beta-adrenoreactivity of erythrocyte membranes (beta-ARM) during two years following renal denervation (RD) in patients with resistant arterial hypertension (RAH) and to determine the relationship of this index with long-term antihypertensive and cardioprotective effectivity of this invasive treatment.Material and methods  The study included 48 patients (mean age, 57.2±8.7 years, 18 men) with RAH on a stable antihypertensive therapy. Averaged daily systolic and diastolic blood pressure (SBP and DBP) and levels of beta-ARM were determined at baseline and in 7 days and 2 years following RD. Measurement of beta-ARM was based on beta-adrenoblocker inhibition of erythrocyte hemolysis induced by exposure to hypo-osmotic environment. The beta-adrenoblocker binds to erythrocyte membrane beta-adrenoceptors to prevent the erythrocyte destruction. Increased values of beta-ARM reflect a decrease in the number of functionally active erythrocyte membrane beta-adrenoceptors associated with long-term sympathetic hyperactivity.Results For two years of follow-up, values of average daily BP decreased from 160.4±16.0 / 88.1±14.6 to 145.3±19.3 / 79.4±13.6 mm Hg. At 7 days, the number of beta-ARM had decreased in the group of RD responders (р=0.028) who at two years had decreased their BP by 10 mm Hg or more, while in the group of non-responders, the number of beta-ARM remained unchanged. At one week, beta-ARM values correlated with changes in SBP and DBP (r= -0.54; р<0.05) and with left ventricular myocardial mass (LVMM) (r= -0.36; р<0.05) at two years of follow-up whereas beta-ARM delta at one week was interrelated with the renin concentration in the long-term (r= -0.44; р<0.05). At two years, the content of beta-ARM was increased in both groups.Conclusion      The decrease in beta-ARM content at 7 days after RD shows the procedure efficacy and allows an expectation of clinically significant decreases in BP and LVMM in the long-term after the surgical treatment. At two years after the intervention, the content of beta-ARM increased, and the BP decrease was apparently due to some other mechanisms.

[Dynamics of adreneractivity after transfer of myocardial infarction: annual observation].

AIM: To study the change in the -adrenergic reactivity of red blood cell membranes in patients during the first year after acute myocardial infarction. MATERIALS AND METHODS: The study included 25 patients with acute myocardial infarction (AMI) who signed informed consent to participate in the study. The erythrocyte membrane -adrenoreactivity index (-ARM) was determined in venous blood samples upon admission to the intensive care unit, one day after admission, 6 and 12 months after the index MI was transferred using the BETA-ARM-AGAT reagent kit (Agat-Med, Russia). RESULTS: According to the results of dynamics assessment of -APM during the first day, patients included in the study were divided into 2 groups. Group 1 (n=14) included patients who had an increase in -APM in the first day, and group 2 (n=21) included patients in whom -ARM either did not change or decreased. At the time of admission to the hospital in the formed groups, there were no differences in the -APM index and clinical and anamnestic characteristics. A day after hospitalization, the value of -APM in group 1 significantly exceeded the same indicator in group 2 (p=0.02). At the periods of 6 and 12 months, the -APM indices in the groups did not differ. In the 2nd group of patients, the progression of chronic heart failure to one or more functional classes (NYHA) was significantly more often compared with the 1st group. CONCLUSION: The study showed that on the first day in patients with AMI, both an increase and a decrease in the activity of the sympathoadrenal system are possible with a further leveling of these differences over the next year. For a group of patients with decreased activity of sympathoadrenal system on the first day, a more unfavorable course of heart failure in the post-infarction period is characteristic.

Comparative tests of quality of bacteriological investigation with Corynebacterium diphtheriae isolation.

The results of comparative experimental studies of identification of nontoxigenic C. diphtheriae strain by three different commercial laboratories are presented. A typical nontoxigenic strain of C. diphtheriae biovar mitis was used. For the studies, three lines of ten-fold dilutions of bacterial culture were prepared, followed by control planting on the medium and counting CFU/ml. In the experiment, tampons were pooled with a 24-hour bacterial culture of a nontoxigenic C. diphtheriae strain. Tampons were provided from three different laboratories - ∑-Transwab® with Ames liquid medium (from the first and second laboratories) and a viscose tampon with coal medium (from the third laboratory). After pooled, tampons were delivered to commercial laboratories. And as a result of the experiment, Corynebacterium spp. was identified in first laboratory (103 CFU/tamp), S. epidermidis (102 CFU/ml) - in second laboratory and nontoxigenic C. diphtheriae biovar gravis - in third laboratory. The study indicates that there is a need to the supervision of bacteriological investigations conducted in various laboratories. This will improve the quality of investigations on diphtheria infection and identify of diphtheria carrier, which is a reservoir of the causative agent of diphtheria, and will contribute to the maintenance of sanitary and epidemiological well-being in our country.

Digital analysis and quantitative assessment of the cervical surface with dysplasia.

The investigation aims - a quantitative assessment of cervical surface changes with digital analysis and computer technologies in dysplasia. Colposcopy was made in 90 women from 21 to 52 years (avr. age 33,9±8,13 y.o.) with mild epithelial dysplasia (CIN1), moderate dysplasia (CIN2), severe dysplasia (CIN3). The algorithm detected indicators which provide the cervical dysplasia classification on pre cytological and pre molecular-genetic patients investigations. The outcome of an algorithm was the identification of the cervix surface condition severity by an objective quantification. The cervical dysplasia type (CIN) was classified as IndGV values. The mild dysplasia (CIN1) had IndGV=8,5, moderate dysplasia (CIN2) - IndGV=13, severe dysplasia (CIN3) - IndGV=15,6. The cervical affected surface area (IndInt) equalled 0,17 in CIN1, 0,19 in CIN2, 0,22 in CIN3. A change severity has a direct relation with a grey color value. It demonstrates quantify classification in digital analysis. The algorithm is used in real-time mode and no requires considerable material outlays. This makes it possible to use an algorithm after clinical examination and predict patient management.

Possibilities of practical application of different culture mediums for laboratory diagnostic of diphtheria.

The purpose of the work is to evaluate the cultural and morphological properties of colonies of clinically significant corynebacteria on culture mediums for the isolation of corynebacteria. The study used 9 culture mediums for the isolation of corynebacteria: a culture medium for the isolation of corynebacteria (Corynebacagar); Tellurite-containing blood agars on base - Culture medium № 1 GRM, Culture agar for the cultivation of microorganisms (GRM agar), Culture medium for determining the sensitivity of microorganisms to antibacterial preparations - AGV, culture agar for the cultivation of dry microorganisms (SPA), Clauberg medium II, Hoyle Medium agar (Oxoid), Blood agar base (Conda), Columbia Agar Base (Conda). The work used 7 test strains of microorganisms from the State collections of pathogenic microorganisms - C. diphtheriae biovars gravis, mitis, intermedius, belfanti and subspecies lausannense, C. ulcerans and C.pseudotuberculosis. Studies were carried out in accordance with MUK 4.2.3065-13 «Laboratory diagnosis of diphtheria infection¼. We describe culture-morphological properties of strains on all tested culture mediums the isolation of corynebacteria after 24 and 48 hours of incubation. Analysis of the results on the growth properties of culture mediums showed that all culture mediums had high sensitivity - from dilution 10-7 for all test strains. Colonies of corynebacteria were visually detected on culture mediums after 19-20 hours of cultivation. When cultivating a suspension of corynebacteria from breeding 10-6 on culture mediums, the number of colonies ranged from 95±5 to 120±10. Conclusion. All culture mediums had differential diagnostic properties that ensure the growth of corynebacteria after the day of incubation.

PCR-based diagnosis of whooping cough in the Russian Federation.

The aim was to determine how often the PCR method is used in different laboratories in Russia. In 2018, we conducted a questionnaire survey in diagnostic laboratories of medical organizations and the Centers of Hygiene and Epidemiology that performed PCR studies to identify microorganisms of the genus Bordetella in all 85 Russian regions. We found that in 2013 the PCR was used in 33 (38.8%) regions, but in 2017 the number of regions increased to 64 (75.3%). During 2013-2017 the study has not been applied in 21 regions. The number of PCR tests performed in the laboratories of medical organizations was significantly different. There has been an increase in the number of tests for the diagnosis of pertussis among people with clinical signs of infection and among contact persons in foci of infection. Compared to the Centers of Hygiene and Epidemiology, in medical organizations the rate of introduction of the PCR was higher. Between 2013 and 2017 the proportion of samples containing DNA B.pertussis decreased, but the proportion of samples containing DNA of other representatives of the genus Bordetella increased. Moreover, in the case of isolation DNA Bordetella spp. clinicians diagnose «Whooping cough, other unspecified organism¼, since there is no information on the species of the pathogen. Thus, in order to improve the diagnosis of pertussis, it is necessary to optimize PCR tests by including target genes that allow to identify of currently relevant DNAs of different representatives of the genus Bordetella.

Erratum: Evolution of π

This corrects the article DOI: 10.1103/PhysRevLett.109.152301.

Fabry Disease in Young Ischemic Stroke Patients in Northern Israel.

INTRODUCTION: The prevalence of Fabry Disease (FD) with cerebrovascular complications varies in different populations. The aim of this study was to estimate the presence of FD among young stroke patients in northern Israel. PATIENTS AND METHODS: We performed a retro-/prospective search for FD in young patients (aged ≤50 years old) admitted to the Department of Neurology due to acute ischemic stroke of any etiology. RESULTS: Overall, 114 patients were examined for FD. Mean age of patients was 40±7.44 years. There were 75 (65.78%) males. FD was found in 4 (3.5%) patients. None of the FD patients had a cryptogenic stroke. CONCLUSION: The results of our study call for a search of FD in young stroke patients of any etiology, and not only among cryptogenic ones.

[Age-dependent changes in adrenoreactivity in experimental postinfarction cardiosclerosis.]

Presently, there is growing interest in studying the activity of the sympathoadrenal system in the aging process of the body and the development of diseases of the cardiovascular system associated with age. The intention of the research is evaluation of ß-adrenoreactivity of erythrocyte membranes (ß-ARM) of intact rats and rats with postinfarction cardiosclerosis at the age of 4, 12 and 24 months: the first group contained twenty 4 months old animals; the second group - twenty 12 months old animals; the third group - twenty 24 months old animals. In the each group were distinguish 10 intact rats and 10 experimental rats, that have postinfarction cardiosclerosis (PICS) modeled by coronary occlusion. The experiments was completed on 60 male-rats. It was shown, that in intact animals the indicator of ß-ARM significantly decreases by 12 months in relation to the group of 4 monthly animals. In the groups of intact animals of 12 and 24 months, there were no significant differences in ß-ARM. The severity of cardiac hypertrophy in animals with PICS was inversely related to age - 4>12>24 months old. Formed PICS in animals aged 4 and 12 months was accompanied by a expressed increase of ß-ARM, in animals 24 months with PICS ß-ARM did not significantly change. Changes in the ß-ARM index characterizing the state of the CAS receptor link under PICS conditions are age-related, in adults 12 months of age with PICS, an increase in ß-ARM relative to the parameter at 4 months. animals indicates compensatory desensitization of ß-adrenergic receptors upon activation of CAS in response to the development of contractile myocardial insufficiency. In the age group of 24 months with the formed cardiosclerosis, compensatory activation of CAS is manifested to a lesser extent.

Peculiarities of Cell Seeding on Electroformed Polycaprolactone Scaffolds Modified with Surface-Active Agents Triton X-100 and Polyvinylpyrrolidone.

We compared the capability of human fibroblasts to populate porous polycaprolactone (PCL) scaffolds modified during fabrication with surface-active agents Triton Ð¥-100 (type 1 scaffold) and polyvinylpyrrolidone (type 2 scaffold). The mean fiber diameter in both scaffolds was almost the same: 3.90±2.19 and 2.46±2.15 µ, respectively. Type 1 scaffold had higher surface density and hydrophilicity, when type 2 scaffold was 1.6 times thicker. The cells were seeded on the scaffolds by the dynamic seeding technique and then cultured in Petri dishes with nutrient medium in a humid atmosphere. During 3-day culturing, no cell release from the matrix was noted. DAPI staining proved the presence of cells in both scaffolds. However, in type 1 scaffold the cells populated the whole thickness, while in type 2 scaffold, the cells were present only in the superficial layer. These findings suggest that PCL scaffolds modified with Triton Ð¥-100 or polyvinylpyrrolidone are not cytotoxic, but the structure of the scaffold treated with Triton Ð¥-100 is more favorable for population with cells.

[The human lectin supersystems possessing probiotic and protective actions.]

The potential of useful for human immunobiological supersystems of lectins (SSL) recognizing carbohydrates and glycoconjugates of molecular or supramolecular protein/(oligo)peptide-containing constituents of biotopes is described. SSL recognize, reversibly bind, delivery to biotopes, orient natural or synthetic polymeric, polyvalent glycoconjugates (imitators of natural glycopolymers) at the cell surface. The key features of SSL are indicated and described. The possibilities of application and prospects of SL of probiotics, complement С4 system and protein hormones (on example of erythropoietins) in prognostics and diagnostics of pathologies, prophylaxis and therapy of diseases and medical biotechnology are evaluated.

Status and problems of bacteriological diagnosis of diphtheria infection in the Russian Federation.

The purpose of the work was to assess the state of bacteriological diagnosis of diphtheria infection in Russia in order to establish possible reasons for the decrease in the release of C. diphtheriae. The Reference Center for Monitoring the Pathogens of Measles, Rubella, Mumps, Pertussis and Diphtheria in 2018 in 85 subjects of Russia conducted a questionnaire of laboratories of medical organizations and the Centers for Hygiene and Epidemiology of Rospotrebnadzor, carrying out bacteriological studies for diphtheria infection. It was found that the number of studies conducted over the five-year period decreased by 1.2 times. The tendency to decrease the number of bacteriological studies for diphtheria is observed in the territories of almost all federal districts. In 99% and 29% of cases, the institutions of the FBUZ Centers for Hygiene and Epidemiology and medical organizations (MO) and use in their work documents regulating bacteriological studies for diphtheria infection. In a number of territories, the list of documents used includes documents that are invalid or do not define such studies. Most organizations use dry tampons when examining for diphtheria, however, 13.1% and 53.4% of FBUZ Centers for Hygiene and Epidemiology and medical organizations (respectively) use commercial transport environments, which does not comply with regulatory documentation. Analysis of the quality of work of bacteriological laboratories showed shortcomings at the stage of preparation of media (use of donor blood, or absence of addition of blood and potassium tellurite), Elek tests (addition of horse serum or absence of serum to the medium), setting of incomplete biochemical series (absence of tests for urease and nitrate reductase), absence of standard control strains, incomplete volume of internal laboratory quality control. Given the continuing circulation of the pathogen in various countries of the world and in our country, as well as the possibility of imported cases of infection from endemic regions, the analysis was aimed at drawing the attention of specialists to the problem of improving the quality of laboratory diagnosis of diphtheria in Russia.

Development of accelerated genodiagnosis method of pertussis and pertussis-like diseases on the basis of mPCR-RT.

The aim of the work was to develop an accelerated genodiagnosis method based on mPCR-RT for the detection DNA of B. pertussis, B. parapertussis, B. holmesii. MATERIALS AND METHODS: The study used 104 strains of microorganisms, of which: 50 strains of B. pertussis, 37 - B. parapertussis, 17 - heterologous species of microorganisms. Assessment of analytical specificity was carried out using DNA strains of various microorganisms with a concentration at least 109 GE / ml. To check the analytical sensitivity we studied a series of serial dilutions of bacterial cultures of the control strains B. pertussis № 143, B. parapertussis № 38b, B. holmesii DSM 13416 with a concentration of 5x109 - 5 µm/ml. RESULTS: Insertion sequences were chosen as diagnostic targets: for B. parapertussis - a specific fragment IS1001, for B. holmesii - a specific fragment hlIS1001, for B.pertussis - a fragment IS481. To develop a genodiagnosis method specific primers were designed and combined into a single multi-primer mixture, the composition of the reaction mixture and the amplification conditions were selected. The analytical sensitivity of the developed method for detecting pertussis and pertussis-like pathogens was 5×101 GE / ml. Verification of the developed methodology of gene diagnostics showed 100% analytical specificity. CONCLUSION: An accelerated genodiagnosis method based on mPCR-RT has been developed, it allows you to identify DNA of B. pertussis, B. parapertussis, B. holmesii, which expands the possibilities of examining patients with suspected pertussis and pertussis-like diseases in order to increase laboratory confirmation of the diagnosis.