ФГБУ «Уральский научно-исследовательский институт охраны материнства и младенчества» Минздрава России, г. Екатеринбург Филогенетический анализ нуклеотидных последовательностей гена uge, детектированного в штаммах Klebsiella pneumoniae Резюме Актуальность. Штаммы Klebsiella pneumoniae могут обладать различными генами факторов вирулентности и вызывать такие нозологические формы, как пневмония, инфекция мочевыделительной системы, абсцесс печени, неонатальный сепсис. Цель работы-провести филогенетический анализ нуклеотидных последовательностей гена uge, детектированного в штаммах K. pneumoniae.

Развитие легких является многостадийным процессом, нарушение которого приводит к формированию респираторных нарушений у новорожденных. Цель исследованияоценить течение неонатального периода недоношенных новорожденных с каналикулярной и саккулярной стадией развития легких. Проведен ретроспективный анализ медицинской документации 46 недоношенных новорожденных с летальным исходом, имевших каналикулярную (n=28) и саккулярную стадии развития легких (n=18). Летальный исход в раннем неонатальном периоде чаще регистрировался у детей с каналикулярной стадией развития легких (75% против 44,5%), причина смерти по патологоанатомическому заключению: инфекция -61,5%, РДС -38,5%. У детей с саккулярной стадией развития более половины умерло на 8-28е сутки (55,6% против 25%), причиной смерти в 100% случаев явилась инфекция. Неонатальный период у всех детей характеризовался развитием респираторного дистресс-синдрома, пневмопатий, внутриутробной инфекцией и неврологической патологией. В первой группе чаще регистрировалась глубокая морфофункциональная незрелость головного мозга. По показателям газового гомеостаза и кислотно-основного состояния в первые часы жизни у всех недоношенных детей наблюдался смешанный и метаболический ацидоз. Гемограмма на 1-е сутки характеризовалась абсолютным лейко-и лимфоцитозом. В показателях красной крови наблюдалось снижение гемоглобина и гематокрита, что потребовало проведения гемотрансфузии в 2,7 раза чаще у новорожденных с каналикулярной стадией развития. На 5-е сутки жизни у всех новорожденных отмечено уменьшение количества тромбоцитов и лимфоцитов. Таким образом, недоношенные дети с различной стадией развития легких имеют похожее течение неонатального периода, однако при каналикулярной стадии развитие патологических процессов стремительнее вследствие большей незрелости ткани легкого; при саккулярной стадии -постепенное нарастание патологических процессов, что диктует необходимость поиска лабораторных диагностических критериев определения степени зрелости легких у новорожденных для разработки персонифицированного подхода интенсивной терапии и возможного повторного введения препаратов сурфактанта. Ключевые слова: недоношенные новорожденные, легкие, преждевременные роды, развитие легких, респираторный дистресс-синдром, неонатальная смертность.

Introduction. Currently, there is insufficient data on the prevalence of Klebsiella pneumoniae strains with virulence factors genes uge and fim among women and newborns. This indicates the need for a study of the prevalence of K. pneumoniae (uge+, fim+) and the degree of heterogeneity of the bacterial population isolated from children and adults.The aim of the study was to perform a phylogenetic analysis of the uge and fim genes of the K. pneumoniae strains.Materials and methods. Total 65 strains of K. pneumoniae isolated from samples of feces, blood, urine, placenta, cervical canal, pharynx, suture of 39 newborns and 24 women were studied. Two blood cultures were obtained from one patient with an interval of two weeks, and two isolates were obtained from the separated cervical canal and suture of one patient. The presence of genes was detected by PCR, nucleotide sequences of the genes were determined by Sanger sequencing.Results. The frequency of detection of the uge gene was 53.8% (35 of 65), fim gene — 23.1% (15 of 65), which indicates a higher prevalence of uge gene strains compared to fim (p < 0.001). The phylogenetic analysis of 18 nucleotide sequences of the uge gene and 4 of the fim gene demonstrated that the strains were distributed in 7 and 4 clusters, respectively. It was established that for, there are No clear clustering by time and place of isolation, patient age, and type of biological material was observed for both uge and fim genes.Discussion. The results of phylogenetic analysis demonstrate the genetic heterogeneity of the studied population of K. pneumoniae, which is confirmed by the wide geography and time variations in detection of the most genetically close bacterial isolates.

In the context of the global spread of the new coronavirus infection, studies aimed at investigating formation of anti-infectious and post-vaccination immunity are of special importance, which is necessary to prevent and reduce morbidity and mortality due to SARS-CoV-2 infection.Purpose: to assess anti-infectious immunity against SARS-COV-2 in various forms of the disease and development of post-vaccination humoral reactions in medical workers of the perinatal center. Materials and methods. A study of blood serum was carried out to assess SARS-CoV-2-specific IgM and IgG antibodies in 119 medical workers recovered after COVID-19, divided into groups based on the disease severity (mild, moderate and asymptomatic), as well as in 62 vaccinated employees, divided into groups according to age. Semi-quantitative measurement of virus-specific antibodies was carried out by ELISA with test systems "SARS-COV-2-IgG-ELISA-BEST" and "SARS-COV-2-IgM-ELISA-BEST".Statistical processing of the research results was carried out using Microsoft Excel 2010 and Statistica 6. Quantitative characteristics were presented as median (ME), lower and upper quartiles (LQ1-UQ3); qualitative parameters – as absolute value and relative number (%). Difference between groups was analyzed by using the χ2 test (qualitative) and the Mann-Whitney U-test (quantitative).Results. The results of the study showed that the majority of employees with a moderate-severe form of SARS-COV-2 had a high level of IgG (PR - a positivity rate of more than 9.0 arbitrary units) 9 months after the disease compared to those who suffered from mild or asymptomatic (83.3% versus 25.8% and 13.3%, p 0.017) infection. The duration of IgG circulation after former illness had no relation to its severity and patient age. The effectiveness of the primary vaccination "Sputnik V" and revaccination with "Sputnik Light" and "KoviVak" was 100% after inoculating the vaccine second component. The lowest level of antibodies after the first vaccination is recorded in persons over 60 years old (1.48 (1.12-3.25 versus PR = 8.48 (5.78-10.11) and 9.27 (5.84-10, 31) arbitrary units, p 0.017)), in comparison with young and middle-age subjects. The speed SARS-COV-2 elimination of IgG at 6, 9 or more months after vaccination depends on relevant initial peak antibody concentration. Subjects who were initially vaccinated with the KoviVac vaccine, IgG was not detected 2 months after vaccination. The protective effect of "Sputnik V", "Sputnik Light", "KoviVac" after re-infection with SARS-COV-2 averages 71.2%. Conclusion. Thus, the results obtained on assessing anti-infectious and post-vaccination immunity against SARS-CoV-2 emphasize the need for further studies on a larger patient cohort, especially in those with asymptomatic infection as well as the elderly subjects.

Relevance A common mechanism of resistance is the synthesis of enzymes that inactivate the antibiotic (AB). Since its discovery at the end of the 20th century, ESBLs have spread widely throughout the world. Representatives of this group of complex protein compounds have common properties of hydrolysis of ß-lactam antibiotics and differ from each other in amino acid sequence. The genetic determinants that determine their synthesis are the bla-CTX-M, bla-SHV, and bla-TEM genes. Aims. To assess the prevalence of bla-CTX-M, bla-SHV, bla-TEM genes in antibiotic-resistant strains of enterobacteria isolated from samples of patients of the perinatal center. Materials & Methods. We studied 135 strains of ESBL-producing enterobacteria isolated during microbiological monitoring from January 01, 2020 to December 31, 2021. Biological material (feces (92), blood (1), urine (5), cervical canal discharge (33), last (4)) came from patients of the perinatal center (43 women and 92 newborns), examined both for clinical indications and during local microbiological monitoring. Species identification of isolated microorganisms was carried out by bacteriological method, detection of antibiotic resistance genes was carried out using real-time PCR. Results. In 87 cases (64.4%), the studied genetic determinants of antibiotic resistance were successfully identified. Of the 26 studied strains of Klebsiella pneumonia, 24 (92.3%) were able to establish the determinants of resistance to AB, 50% of which are represented by bla-CTX-M. Out of 66 Escherichia coli, 59 (89.33%) had a genetic profile of antibiotic resistance, which was also dominated by bla-CTX-M (67.80%). However, in the Enterobacter cloacae group, it was possible to determine the presence of the studied genes only in 8.10% of cases. K. pneumoniae carrying three antibiotic resistance genes bla-CTX-M, bla-SHV, bla-TEM was isolated once from a sample of the cervical canal of a woman aged 27 years. E. coli strains carrying the CTX-M gene were isolated from 3 mother-child pairs out of 135 cases: in mothers – from the discharge of the cervical canal in the third trimester of pregnancy, in newborns – from feces in the first week of life, which can indicate possible transmission of an AB-resistant microorganism either in utero or through the birth canal. Conclusions. Thus, in the course of the study, determinants of antibiotic resistance of enterobacteria isolated from patients of the perinatal center were identified in 64.4% of cases. The dominant gene leading to the occurrence of AD among ESBL is the bla-CTX-M gene. The number of strains with an established genetic profile of resistance to AB is less among Enterobacter cloacae (p<0.001). In order to study the genetic profile in more detail and to identify the mechanisms for the formation of antibiotic resistance, it is necessary to expand the detection of observed genes