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Introduction: Puumala virus (family Hantaviridae , genus Orthohantavirus ) is distributed in most regions of the European part of Russia. However, information about its genetic variants circulating on the territory of the Central Federal District is extremely scarce., Materials and Methods: Rodents' tissue samples were tested after reverse transcription by PCR for the presence of hantaviral RNA. The amplified fragments of the L segment were sequenced by the Sanger method. For two samples, sequences of all three segments were obtained using the NGS method. Phylogenetic trees were built in the MEGA-X software., Results: Puumala virus was found in six samples. Based on the phylogenetic analysis of sequences of three segments, the obtained genetic variants belong to the sublineage previously designated as W-RUS., Conclusion: A genetic variant of the Puumala virus, belonging to the subline W-RUS, circulates on the territory of the Volokolamsk district of Moscow region.

Introduction: Pestiviruses and viruses of the Herpesviridae family are widely distributed among different species of ungulates, but the main information about these pathogens is related to their effect on farm animals. Data on detection of bovine viral diarrhea virus (BVDV) and bovine herpes virus (BoHV) in wild ungulates reported from different countries in recent years raises the question of the role of wild animals in the epidemiology of cattle diseases., Aim of Work: To study the prevalence of herpesviruses and pestiviruses in the population of wild artiodactyls of the Moscow region., Materials and Methods: Samples of parenchymal organs and mucosal swabs from 124 wild deer (moose and roe deer) shot during hunting seasons 20192022 in Moscow Region were examined by PCR, virological and serological methods for the presence of genetic material and antibodies to bovine infectious rhinotracheitis and viral diarrhea., Results: BVDV RNA was found in a sample from one moose, BoHV DNA was detected in samples from three roe deer and two moose shot in the Moscow region. Seropositive animals were of different sex and age, the total BoHVs and BVDV seroprevalence rates in wild artiodactyls were 46 and 29%, respectively., Conclusion: Wild ruminant artiodactyls of the Moscow Region can be a natural reservoir of BoHV-1, and this must be taken into account when planning and organizing measures to control the infectious bovine rhinotracheitis. Cases of BVDV infection in wild artiodactyls are less common, so more research is needed to definitively establish their role in the epidemiology of this disease in cattle.

Introduction: SARS-CoV-2, a severe acute respiratory illness virus that emerged in China in late 2019, continues to spread rapidly around the world, accumulating mutations and thus causing serious concern. Five virus variants of concern are currently known: Alpha (lineage B.1.1.7), Beta (lineage B.1.351), Gamma (lineage P.1), Delta (lineage B.1.617.2), and Omicron (lineage B.1.1.529). In this study, we conducted a molecular epidemiological analysis of the most prevalent genovariants in Moscow and the region. The aim of the study is to estimate the distribution of various variants of SARS-CoV-2 in Moscow city and the Moscow Region., Materials and Methods: 227 SARS-CoV-2 sequences were used for analysis. Isolation of the SARS-CoV-2 virus was performed on Vero E6 cell culture. Sequencing was performed by the Sanger method. Bioinformatic analysis was carried out using software packages: MAFFT, IQ-TREE v1.6.12, jModelTest 2.1.7, Nextstrain, Auspice v2.34., Results: As a result of phylogenetic analysis, we have identified the main variants of the virus circulating in Russia that have been of concern throughout the existence of the pandemic, namely: variant B.1.1.7, which accounted for 30% (9/30), AY.122, which accounted for 16.7% (5/30), BA.1.1 with 20% (6/30) and B.1.1 with 33.3% (10/30). When examining Moscow samples for the presence of mutations in SARS-CoV-2 structural proteins of different genovariants, a significant percentage of the most common substitutions was recorded: S protein D614G (86.7%), P681H/R (63.3%), E protein T9I (20.0%); M protein I82T (30.0%), D3G (20.0%), Q19E (20.0%) and finally N protein R203K/M (90.0%), G204R/P (73.3 %)., Conclusion: The study of the frequency and impact of mutations, as well as the analysis of the predominant variants of the virus are important for the development and improvement of vaccines for the prevention of COVID-19. Therefore, ongoing molecular epidemiological studies are needed, as these data provide important information about changes in the genome of circulating SARS-CoV-2 variants.

Introduction: The study of the intensity of humoral immunity to the measles virus (Paramyxoviridae: Morbillivirus) (MV) remains relevant due to the ongoing registration of the cluster cases of measles (including nosocomial cases).The aim of the study was to analyze correlation between the intensity of the humoral immunity to measles virus and the measles incidence in different age groups of the Moscow residents., Material and Methods: A simple analytic study was carried out under the framework of measles prevention improvement program conducted in the subjects of the Russian Federation. The study included the analysis of the reasons for the increase in the proportion of seronegative individuals and the comparison of the results of seromonitoring with the data of age-related measles incidence. Throughout 2018, total 1,855 blood serum samples obtained from employees of a large hospital center in Moscow who were previously vaccinated against measles were tested for IgG antibodies (ABs) to MV. The subjects were divided into 11 groups according to their age. Statistical analysis of the obtained data was performed using nonparametric criteria., Results and Discussion: The proportion of seronegative individuals among the staff of the medical facility decreased with age, from 38.5% in persons aged 19-23 to 0% among employees aged 64 and over. Persons aged 19-43 years were the most susceptible to measles infection, which was confirmed also by the highest incidence rate at these age groups (from 6.93 to 14.03 per 100,000 population). The age group most protected against measles was those over 64 years of age, who were all seropositive and had the lowest rates of incidence. This is probably due to the fact that by this age most of them have had a measles infection and have formed a sufficient immune response. At the same time, younger individuals living in the post-elimination era have only vaccine-induced ABs to MV, the level of which declines rapidly to non-protective values in the absence of boosterization due to the exposure to the wild-type strain of the pathogen., Conclusion: The monitoring of the MV-specific IgG- AB levels is required for employees in the medical institutions under the age of 35. Administration of the single dose of the appropriate vaccine is recommended for seronegative individuals.

Introduction: Since the outbreak of the COVID-19 pandemic caused by SARS-CoV-2 novel coronavirus, the international community has been concerned about the emergence of mutations altering some biological properties of the pathogen like increasing its infectivity or virulence. Particularly, since the end of 2020, several variants of concern have been identified around the world, including Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), and Delta (B.1.617.2). However, the existing mechanism of detecting important mutations are not always effective enough, since only a relatively small part of all pathogen samples can be examined by whole genome sequencing due to its high cost., Material and Methods: In this study, we have designed special primer panel and used it for targeted highthroughput sequencing of several significant S-gene (spike) regions of SARS-CoV-2. The Illumina platform averaged approximately 50,000 paired-end reads with a length of ≥150 bp per sample. This method was used to examine 579 random samples obtained from COVID-19 patients in Moscow and the Moscow region from February to June 2021., Results: This study demonstrated the dynamics of distribution of several SARS-CoV-2 strains and its some single mutations. It was found that the Delta strain appeared in the region in May 2021, and became prevalent in June, partially displacing other strains., Discussion: The obtained results provide an opportunity to assign the viral samples to one of the strains, including the previously mentioned in time- and cost-effective manner. The approach can be used for standardization of the procedure of searching for mutations in individual regions of the SARS-CoV-2 genome. It allows to get a more detailed data about the epidemiological situation in a region.

Introduction: Measles remains an urgent problem in Russian healthcare. Despite the ongoing vaccination, there is an increase in the incidence of measles. Prevention of measles is particularly important in high-risk groups, as well as among healthcare professionals to prevent hospital-acquired outbreaks of infection. The duration of post-vaccination immunity during the elimination of measles has not been sufficiently studied, so often people who  have had measles in childhood or have 1-2 vaccinations against the disease lose their protective antibodies with age in the absence of natural boosterization.Goals and objectives. To study the intensity of specific immunity to measles in employees of the maternity unit., Material and Methods: The study involved 271 employees of the maternity unit aged 21 to 93 years (262 serum samples). The level of IgG antibodies (Ab) to the measles virus in the blood serum was studied by ELISA using a standard set of reagents for the quantitative determination of IgG by «VECTOR-BEST». The result was  considered negative if the concentration of IgG to the measles virus in tested sample was ≤ 0.18 IU/ml and  positive - if > 0.18 IU/ml. Results. The number of seronegatives ranged from 0% to 30.8% in female employees with its maximum at age of 31-35 years. The lowest proportion of seronegative and the highest proportion of seropositive women were observed among the elderly, > 60 years., Discussion: There is a marked tendency for an increase of the proportion of persons with average Ab levels with age and a decrease of the proportion of persons with low Ab levels. The percentage of seronegative women among employees exceeded the recommended level, which makes it possible for an nosocomial outbreak when an infection is introduced., Conclusion: The authors recommend that serological testing for the intensity of the immune response against measles should be included in the standard of the pre-vaccination screening for adults.

Introduction: The novel SARS-CoV-2 coronavirus, which emerged at the end of 2019 and caused a worldwide pandemic, triggered numerous questions about the epidemiology of the novel COVID-19 disease and  about wellknown coronavirus infections, which used to be given little attention due to their mild symptoms., The Purpose: The routine screening-based multiyear retrospective observational study of prevalence and circulation patterns of epidemic-prone human coronaviruses in Moscow., Material and Methods: The real-time polymerase chain reaction was used to detect RNA of human coronaviruses (HCoVs) in nasal and throat swabs from 16,511 patients with an acute respiratory infection (ARI), aged 1 month to 95 years (children accounted for 58.3%) from January 2016 to March 2020, and swabs from 505 relatively healthy children in 2008, 2010 and 2011. Results. HCoVs were yearly found in 2.6-6.1% of the examined patients; the detection frequency was statistically higher in adults than in children, regardless of sex. At the height of the disease incidence in December 2019, HCoVs were detected in 13.7% of the examined, demonstrating a two-fold increase as compared to the multi-year average for that month. The statistical frequency of HCoV detection in ARI pediatric patients under 6 years was significantly higher than in their healthy peers (3.7 vs 0.7%, p = 0.008)., Conclusion: HCoVs circulate annually, demonstrating a winter-spring seasonal activity pattern in the Moscow Region and reaching peak levels in December. Over the years of observation, the HCoV epidemic activity reached maximum levels in December 2019-February 2020 and decreased in March to the multi-year average. Amid a growing number of SARS-CoV-2 cases imported to Moscow in March 2020, the HCoV detection frequency dropped sharply, which can be explained by the competition between different coronaviruses and by the specificity of HCoV detection with the diagnostic test kit used in this study.

The purpose of the study is to analyze patterns demonstrated by the COVID-19 epidemic process in a megacity during the increase, stabilization and reduction in the incidence, and to evaluate the effectiveness of the epidemic prevention measures., Materials and Methods: The comprehensive study incorporating epidemiological, molecular genetic and  statistical research methods was conducted to analyze the spread of SARS-CoV-2 in Moscow during the COVID- 19 pandemic., Results and Discussion: It was found that the exponential growth in COVID-19 cases was prevented due to the most stringent control and restrictive measures deployed in Moscow to break the chains of SARS-CoV-2 transmission and due to people who were very disciplined in complying with the self-isolation rules. The  analysis of the dynamics in detection of new COVID-19 cases showed that in a megacity, the impact of social distancing and self-isolation would become apparent only after 3.5 incubation periods, where the maximum length of the period is 14 days. It was discovered that the detection frequency of SARS-CoV-2 RNA in relatively healthy population and its dynamics are important monitoring parameters, especially during the increase and stabilization in the COVID-19 incidence, and are instrumental in predicting the development of the epidemic situation within a range of 1-2 incubation periods (14-28 days). In Moscow, the case fatality rate was 1.73% over the observation period (6/3/2020-23/6/2020)., Conclusion: The epidemiological analysis of the COVID-19 situation in Moscow showed certain patterns of the SARS-CoV-2 spread and helped evaluate the effectiveness of the epidemic prevention measures aimed at  breaking the routes of transmission of the pathogen.

Aim: Study manifestations of epidemic process during acute intestinal infections to establish reasons of low effectiveness of the prophylactic measures carried out and evaluation of the role of rotavirus infection in general disease structure of intestinal infections of unknown etiology., Materials and Methods: Data on morbidity of acute intestinal infections of population of Moscow were analyzed. Hospitalized patients with acute intestinal infections were examined using real-time PCR method test-systems of laboratory of molecular virology of Mechnikov RIVS with subsequent typing., Results: Evaluation of multi-year manifestations of epidemic process of morbidity of acute intestinal infections in Moscow has shown, that the cumulative morbidity does not have a tendency of reduction, because the proportion of infections with undeciphered etiological factors is almost 80% of the total aggregate morbidity. The proportion of rotavirus infection in total morbidity of AII of established etiology increased from 53.2 in 2004 to 82.6% in 2014. Morbidity in childrenwith rotavirus infection is 6 times higher than morbidity in adults., Conclusion: The results obtained give evidence on the necessity of carrying out specific prophylaxis against viral intestine infection, mostly of rotavirus and norovirus infections. The highest effect should have been expected from use of a bi-vaccine, development of which seems quite an actual problem.

Aim: Characterize spectrum of antibiotics resistance of Acinetobacter baumannii strains, isolated from patients of 8 surgical and reanimation departments of 3 medical institution of Moscow, and determine molecular-genetic mechanisms of stability of their carbapenem-resistant forms., Materials and Methods: 95 strains of A. baumannii, isolated from patients of reanimation and surgical departments of Moscow in 2012-2014, were studied. Sensitivity of strains to antibiotics was tested phenotypically according to recommendations of EUCAST. The presence of VIM, IMP, OXA-23, OXA-40, OXA-48, OXA-58 and NDM genes in the studied strains was determined by polymerase chain reaction in real time., Results: 86.3% of strains turnedout to be non-sensitive to carbapenems, sensitive--13.7%. 80.0% of strains were non-sensitive to gentamicin, 80.0% of strains--to netilmicin, 94.7% of strains--to ciprofloxacin 2.1%--to colistin. 91.6% of isolates have shown non-sensitivity to members of 2 and more classes of antibiotics, 78.9% of strains--to members of 3 classes. 2 strains were panresistant, 4.2% (4/95) of the isolates were sensitive to all the classes of antibiotics. Metallo-β-lactamases were not detected. Genes of carbapenemases (OXA-23 and/or OXA-40) were detected in 85.3% (81/95) of strains, characterized phenotypically as non-sensitive to carbapenems., Conclusion: The results obtained shown an increase of resistance to carbapenems and multiple resistance in clinically significant strains of A. baumannii. Resistance to carbapenems is associated with OXA-23 and OXA-40 genes. The conclusions allow to justify perspectives of introduction of technologies of molecular-genetic testing of antibiotics resistance.

Aim: Evaluation of the role of contemporary methods in epidemic control of brucellosis outbreaks among employees of auxiliary facilities in Moscow zoo nursery., Materials and Methods: During 2003-2013, biannually, sera from more than 200 employees of the nursery, that work during periods of epizootics of small and large cattle in nursery auxiliary facilities, were studied. A complex of laboratory methods for brucellosis was used: variations of traditional serologic agglutination method in tubes--Wright's reaction (WR), on glass--Huddleston's reaction, Coombs' anti-globulin reaction; enzyme immunoassay with immune-dominant S-LPS; realtime polymerase chain reaction with primers based on genus target of BCSP31 protein gene., Results: After eradication of sheep brucellosis in 2003, the percentage of nursery facility employees, that react positively to brucellosis, has decreased from 42.7% to 15.9-17.2% in 2005-2006. RT-PCR detected human infection during epizootics 5.8 times more effectively compared with EIA. The repetition of the brucellosis epizootics in 2007 and 2009 among large cattle and a 2-year yak had initiated a rise in infection rate among employees, that had not previously (2003-2006) reacted to brucellosis. Acute clinical forms of brucellosis were not detected in 2012-2013 and antibody titers in EIA in previously infected employees have decreased in the absence ofbrucellosis epizootics in the nursery. 30 employees, infected with brucellosis causative agent, were detected for the entire period of examination, among those 10 individuals had developed clinical forms of brucellosis (6--acute, 4--chronic)., Conclusion: Epidemic control for 11 years, based on a complex of laboratory methods, allowed to register infection and outbreaks of brucellosis in the nursery employees during epizootics of 2003, 2007 and 2009. RT-PCR--effective method of detection of infection in humans during brucellosis periods in the nursery. EIA--sensitive method during chronic forms of brucellosis in the periods between and after epizootics compared with RT-PCR. A variation of a traditional serologic method of diagnostics has shown a lower sensitivity, informativity with a larger duration of analysis compared with RT-PCR and EIA.

The rhinoviruses and coronaviruses are the most common causative agents of the acute upper respiratory tract infection in humans. They include several species that vary in the pathogenicity, some causing severe respiratory tract diseases. In this work, the species prevalence of rhinoviruses and coronaviruses was studied in 92 virus-positive clinical patients that were collected at the area of the Moscow region during the period from 2007 to 2012. Using the real-time PCR the virus circulation has been established for all species common in humans, including three rhinoviruses, HRV A, HRV B, and HRV C, and four coronaviruses, HCoV-NL63, HCoV-229E, HCoV-OC43, and HCoV-HKU1. For eight patients, the identity of the rhinoviruses, including 4 cases of HRV-C, 3 cases of HRV-A, and a single case of HRV-B, was corroborated using partial sequencing of the 5 non-coding regions and phylogenetic analysis. The viruses of HRV-C, HCoV-NL63, and HCoV-OC43 were prevalent in children with severe respiratory diseases.

The goal of this work was to analyze the antigenic structure of the hemagglutinin (HA) of the pandemic influenza virus A(H1N1)pdm09 using monoclonal antibodies (MAbs) and to develop a sandwich ELISA for identification of pandemic strains. Competitive ELISA demonstrated that 6 MAbs against HA of the pandemic influenza A/ IIV-Moscow/01/2009 (H1N1)pdm09 virus identified six epitopes. Binding of MAbs with 22 strains circulating in Russian Federation during 2009-2012 was analyzed in the hemagglutination-inhibition test (HI). The MAbs differed considerably in their ability to decrease the HI activity of these strains. MAb 5F7 identified all examined strains; MAbs 3A3 and 10G2 reacted with the majority of them. A highly sensitive sandwich ELISA was constructed based on these three MAbs that can differentiate the pandemic influenza strains from the seasonal influenza virus. The constancy of the HA epitope that reacts with MAb 5F7 provides its use for identification of the pandemic influenza strains in HI test. MAbs 3D9, 6A3 and 1E7 are directed against the variable HA epitopes, being sensitive to several amino acid changes in Sa, Sb, and Ca2 antigenic sites and in receptor binding site. These MAbs can be used to detect differences in HA structure and to study the antigenic drift of the pandemic influenza virus A(H1N1)pdm09.

Aim: Study phylogenetic interconnections of HIV-1 subtype A and B variants circulating in Novosibirsk region (NSR)., Materials and Methods: 268 HIV-1 variants isolated in 2007 - 2010 from blood samples of HIV infected patients in NSR, Samara, Congo and Moscow. HIV-1 variant genotyping was performed by analysis of 1.3 kb long pol gene nucleotide sequences. Phylogenetic analysis of nucleotide sequences was carried out by program Mega version 4.1 by constructing phylogenetic trees by nearest neighbor method. Nucleotide distances were calculated by Kimura method., Results: The studied HIV-1 subtype B variants form separate phylogenetic groups with a low HIV-1 nucleotide sequence homology level combined based on territorial principle and/or time of HIV infection in a territory but not possessing interconnection with a specific population risk group. Subtype A HIV-1 is a fairly homogenous monophyletic group. Phylogenetic differences during studies of HIV-1 isolated from risk group patients - injection drugs users and individuals infected through sexual contacts were not detected. HIV variants isolated from patients infected in Moscow and Samara generally grouped with HIV variants circulating in the European part of Russia., Conclusion: An independent circulation of genetically separate HIV-1 subtype B groups is observed on the territory of siberian region which is a result of multiple independent introductions of distant variants of the virus. The confirmed limited spread of this HIV-1 genetic variant with a subsequent territorial separateness creates a possibility of formation of genetically different virus populations. The studies of subtype A viruses performed confirm the high level of homogeneity detected earlier in other Russia territories of HIV-1 belonging to this genetic variant. Monophyly of subtype A HIV variants is explained by imposition of 2 factors - territorial mobility of the population inside the country and lack of specific transmission routes for HIV-1 subtype A.

Aim: Study of taxonomical structure ofwound infection agents, prevalence of mixes, and detection of character of their possible connection with the results of various microorganisms population interaction in septic wounds., Materials and Methods: A microbiological study of material from patients with wound infection (WI), 582 of those were cured in reanimation and intensive therapy departments (RITD; group 1) and 1455 - in surgical departments (SD; group 2), was performed. Taxonomic membership and ability to coexist was determined in 4129 microorganisms strains. Etiological role of the agents was evaluated by using values of consistency rate (CR). Species that were present in more than 50% of samples were considered consistent, in 25 to 50%--additional, and in less than 25%--random. Frequency rates (FR) were also determined, that is the fraction of a certain species (genus) of the microorganism (in %) from all the isolated cultures that correspond to 100%. For the determination of the significance of individual species of the agent in the structure of mixed microorganism populations, FR - their fraction (%) in mixed population from the number of strains of this species that correspond to 100% - was calculated., Results: A significant part of the microorganisms strains, more frequently in reanimation department (65.5%), caused wound suppuration in populations mixed with other species of the agents. In reanimation and surgical departments consistent species of wound infection agents were not detected. A leading etiological role of Staphylococcus aureus (FR 19.2% and 23.9%) was determined, and FR of S. aureus strains in mixes was 64.6% in RITD and 46.8% in SD. The parameters ofotheragents of WI in the comparison groups were similar. However FR among mixes in RITD were significantly higher for streptococci that do not belong to S. pyogenes species (72,5%), and also nonfermentative microorganisms (67,2%), and in SD - in Klebsiella pneumoniae mixes. For agents of wound infection especially in RITD, low species diversity was characteristic and the number of mixes variants is significantly higher. In RITD mixed infections develop more frequently, and the ecological community of microorganisms reaches higher values than in SD., Conclusion: During the analysis of microbiologi-cal data in RITD and SD general patterns and specific features of taxonomical structure, prevalence of mixed populations and character of their ecological community in wound infection was determined.

Aim: Studies of hepatitis C virus (HCV) genotype and subtype structure in patients with chronic hepatitis C in 3 regions of the Central federal district of Russia., Materials and Methods: Hepatitis C virus genotype and subtype structure was determined in patients with chronic HCV infection in Moscow (1993 - 1995 and 2005), Moscow region (2008) and Vladimir region (1993 -1995, 2005-2007). HCV genotype was determined by using A. Widell et al. (1994) technique, PCR (AmpliSens diagnostic kits), Genotype C test system., Results: In all studied regions and during all the time periods the first position in rating belonged to HCV 1b subtype. In 1993 - 1995 and 2005 - 2007 period changes in HCV genotype and subtype structure were registered that consisted of relative weight of 1b subtype decrease and 3a subtype increase. Subtype 1b in females with chronic hepatitis C was registered more often than in males. In Vladimir region 3a subtype in males was detected more often than in females. In males older than 30 years the first rating position belongs to 1b subtype and in males younger than 30 years--subtype 3a. In females older than 30 years in Moscow region and Vladimir region, as well as in females younger than 30 years in Vladimir region subtype 1b was detected more often, while in Moscow region HCV subtypes 1b and 3a were detected with the same rate of 47.6%., Conclusion: Currently there is an urgent need to include mandatory monitoring of hepatitis C virus genetic variants into the system of hepatitis C epidemiologic control in Russia. This approach will allow for a significant increase in quality of hepatitis C serological diagnostics, and can be used in the prognosis of evolution of the epidemic process of this disease.

Aim: Analysis of clinical cases of tick-borne spotted fever (TSF) group rickettsiosis in 2005 - 2010., Materials and Methods: General clinical, biochemical and serological parameters were determined in 10 tick-borne spotted fever group rickettsiosis patients who had visited various geographical regions of the World., Results: TSF group rickettsiosis diagnostic criteria, optimal serological diagnostics timing were determined. Possible diagnostic errors, features of serological diagnostics and antibacterial therapy of this nosologic form are discussed., Conclusion: Indication for TSF examination are primarily epidemiologic including tick attachment indication and clinical data. Serological studies are positive only in 3 - 4 weeks after the onset of the infection and thus can not be used for early diagnostics.

Aim: Genetic and antigenic characterization of Neisseria meningitidis strains isolated during meningococcal infection outbreaks from individuals in contact with patients with generalized form of meningococcal infection., Materials and Methods: Strains obtained in 2007 - 2009 in Moscow during examination of individuals that were in contact with patients during meningococcal infection outbreaks were analyzed. Multilocus sequence typing, genetic subtyping and typing of VR fragment (FetA) techniques were used., Results: Data regarding investigated strains were submitted to the database at http://pubmlst.org/neisseria/. Previously undescribed sequence types were found in 12 strains, sequence-type could not be determined in 2 strains, 2 strains lacked VR fragment (FetA). Serogroup A meningococci had "P1.5-2,10: F3-5" antigenic profile and belonged to ST-75 and ST-3349 sequence-type, these data does not support the emergence of epidemically significant strains in the territory under surveillance. All typed serogroup C strains and 1 serogroup B strain are of "ST-41/44 complex/Lineage 3" clonal complex. Subtypes of serogroup C meningococci strains match subtypes of strains that cause generalized forms of infection, while serogroup B strains isolated from the carriers and strains isolated from the patients had different antigenic profiles. Ungrouppable strains had notably higher level of genetic and antigenic diversity: only 6 of 16 strains (37.5%) could be sequence-typed using earlier data, all these strains are of clonal complex "ST-53 complex" that consists mostly of strains isolated from the carriers. CONCLUSION. Ratio of meningococci population circulating in Moscow and subpopulation capable of causing generalized form of meningococcal infection (GFMI) is different for meningococci of various serogroups. Ungrouppable strains isolated from the carriers are highly different from strains causing GFMI.

Aim: To assess rate and level of contamination by Legionella pneumophila of cooling water systems in industrial facilities as well as hot water supply systems of administrative buildings in Moscow region., Materials and Methods: Cooling water systems of 8 industrial facilities and hot water supply systems of 12 administrative buildings or complexes located in Moscow or Moscow region were examined. Samples of water, washes and biofilms were studied by bacteriologic methods and RT-PCR. Results. Significant level of contamination of water systems by L. pneumophila was revealed in examined objects. Rate of contamination of cooling water systems in industrial facilities was 70%. The agent was detected in stagnant, end-capped, and rarely used segments of all hot water supply systems during decrease of water temperature to 36-52 degrees C. Visual detection of natural biofilms on the object correlated with high concentration of L. pneumophila in water samples. In some cases, associations of L. pneumophila with Pseudomonas aeruginosa were detected, including water samples from supply systems of 2 healthcare facilities., Conclusion: Obtained results confirm the importance of implementation of modem concept of legionellosis prevention in our country, based on regular quantitative monitoring for Legionella in potentially dangerous water objects and conduction of preventative measures if contamination exceeds acceptable level.

Aim: Comparative analysis of structure of tcfA gene coding tracheal colonization factor of Bordetella pertussis strans isolated in Moscow from patients with pertussis., Materials and Methods: Ninety-seven strains of B. pertussis isolated in different periods of pertussis infection epidemic process (1948 - 1989--from collection of Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology; 1990 - 2007--isolated in Moscow from patients with pertussis) were studied. Primers for amplification of tcfA gene region with size 945 n.p. were used. Amplicons obtained in PCR were used for sequencing. Nucleotide sequences of tcfA gene types of B. pertussis strains were matched to EMBL/GenBank database., Results: Sequencing of tcfA gene fragments revealed two sequence variants. Ninety-six of 97 studied B. pertussis strains had the same sequence variant--variant 1. The one strain was characterized by other nucleotide sequence--variant 2, which differed from variant 1 by presence of insertion g in position 396 that led to reading frame shift., Conclusion: The structure of tcfA gene circulating population of B. pertussis strains is homogenous and is characterized by presence of "vaccine" allele dominating in majority of countries in the world.

The Moscow Region is one of the HIV-1-affected subjects of the Russian Federation; there were 34613 HIV-1-infected subjects as of October 31, 2009. To characterize the molecular epidemiology of HIV-1 in the Moscow Region, the investigators obtained and studied HIV-1 variants from 61 infected subjects of the region, who were major risk groups: intravenous drug users (IDUs) and hetero- and homosexually infected persons. Genetic analysis of HIV-1 variants was carried out by sequencing the gag genes (729 nucleotides in length, including full-length protein p17 and partial p24) andlor env (270 nucleotides in length, V3 region) with further phylogenetic analysis. The findings demonstrated that HIV-1 subtype A variants are dominant in the Moscow Region and detectable in 93.5% of IDUs and 100% of heterosexually infected persons. Phylogenetically (and accordingly epidemiologically) unrelated HIV-1 subtype B strains were revealed in 4 patients, including 2 IDUs.

Aim: Genetic characterization of 37 strains and CSF samples containing DNA of Haemophilus influenzae type b isolated in Moscow during 2007-2009., Materials and Methods: Multilocus sequence-typing method was used and also variant of method for capsule type determination was approbated., Results: Ten sequence types, of which 7 were described in previous studies and 3 were revealed for the first time during this work, were detected in studied sample. ST-6 and ST-92 were the most frequently detected--9 strains (24%) of Hib belonging to each sequence type were revealed. All detected sequence types, except one, belong to clonal complex "ST-6" ("A1/A2"). Obtained data were compared with results of typing of Hib strains isolated in Moscow in 1999-2001. Genetic changes in studied population of Hib are characterized by decreased proportion of Hib belonging to ST-6 (from 54% to 24%) and increased number of sequence types belonging to clonal complex "ST-6" differing from ST-6 on more than one locus of allelic profile (from 2 types [2 strains, 5.4%] to 5 types [9 strains, 24%])., Conclusion: In 2007-2009, number of Hib strains with sequence type ST-95 (7 strains, 19%), which is typical for strains circulating in Russia, is markedly increased. Capsule type I was detected in 32 (86.5%) of studied strains, whereas capsule type II--in 5 (13.5%) of studied strains. Capsule type II was detected only in Hib strains with ST-80 sequence type.

Aim: Analysis of influenza morbidity and mortality including delayed mortality as well as study of the association between increased mortality in elderly and influenza incidence., Materials and Methods: Epidemiologic methods of research as well as reporting forms No. 2 and C51 for Moscow for the period 1999-2005 were used. Analysis was performed for 28,801,565 cases of influenza illness as well as for 41,310 fatal cases, of which 6048 cases due to cardiovascular and respiratory diseases were extracted. Retrospective epidemiologic analysis of perennial and within-year incidence of influenza and mortality due to mentioned diseases in Moscow was performed. Correlation analysis was done using Spearman rank correlation coefficient and Pearson correlation coefficient., Results: Delayed mortality concept was introduced. Existence of delayed mortality after influenza illness in persons >60 year old with ischemic heart disease, acute myocardial infarction and cerebrovascular disease was proved. It was established that vaccination against influenza in persons aged >60 years with underlying cardiovascular diseases during epidemic season could prevent 246 deaths each year., Conclusion: Delayed mortality was detected only in patients with cardiovascular diseases, whereas in patients with respiratory diseases direct complications due to influenza developed. Influenza immunization of elderly persons could yield marked economic effect as cost of treatment of one case of acute myocardial infarction or attack of cerebrovascular disease is significantly higher than cost of vaccination.

Herd immunity against poliomyelitis was studied in 1391 children and adolescents from 10 towns of the Moscow Region. It was ascertained that the values of herd immunity against poliomyelitis virus type 1 were high everywhere and those of herd immunity against poliomyelitis virus type 2 were high and very high in 9 towns and below the WHO minimum levels (80%). The values of herd immunity against poliomyelitis virus type 3; they were lower than the required minimum in 2 towns and very low in 2 other towns arouse alarm. The study of strain-specific antibodies to vaccine-derived and wild polioviruses has demonstrated that wild poliomyelitis viruses have not circulated in the areas of the examinees in the past decade.

Aim: To study pathogenic characteristics of B. pertussis strains isolated from patients during different periods of pertussis infection epidemic process., Materials and Methods: Strains of B. pertussis isolated in Moscow during 1967 - 1971, 1980 - 1985, and 2001 - 2005 were studied. Nutrient media: Bordet-Gengou blood agar, casein-charcoal agar., Animals: mice - F1 hybrids (CBA x C57BL6). Pathogenic characteristics of strains were studied by assessment of virulence (LD50), leukocytosis-stimulating (LS units) and histamine-sensitizing (HSD50) activities of cultures. Genotyping was performed using standard equipment and reagents for DNA isolation, amplification, sequencing and detection of results., Results: On the sample of 164 strains, pathogenic and genotypic characteristics of B. pertussis populations circulated during 1967 - 1971, 1980 - 1985, and 2001 - 2005. Majority of B. pertussis strains isolated in 1967 - 1971 and strains circulated during current phase of epidemic process were virulent (80.75% and 81.8% respectively) and had significant leukocytosis-stimulating and histamine-sensitizing activity, whereas strains isolated from patients with pertussis in 1980 - 1985 characterized by lower virulence and toxicity. Genotyping showed strains carrying "non-vaccine" allele ptxA1, which emerged in the middle of 1970s, totally displaced strains with "vaccine" alleles ptxA2 and ptxA4., Conclusion: Adaptive changes of B. pertussis driven by increased vaccination coverage involve both ptxA gene and pathogenic characteristics of infectious agent in the range of genotypically homogenous population with domination of strains, which have high levels of virulence and toxicity.

Aim: To study etiologic role of microorganisms during wound infection and elucidation of their populations interaction in patientswith burns., Materials and Methods: 2137 patients with wound infection treated in burn center were studied. 3179 samples of wound exudates and 6501 strains of microorganisms were studied. Assessment of microorganism's etiologic role was carried out using results of calculation of various statistic indexes, including indexes of ecological similarities of microorganisms in the wound (indexes of contamination, prevalence or persistence, incidence). Ecological similarity of wounds' microflora was determined by calculation of the coefficient Kj., Results: It was established that Staphylococcus aureus is the most common cause of mono- and mixed wound infections. It was determined that Kj value for common agents points to antagonism or associative interaction. During particular time periods of the study values of Kj significantly differed and depended from species. Specifically, for Pseudomonas aeruginosa the values of Kj were equal or exceeded 30% during several years. It was the first time when correlation between persistence and ecological similarity indexes was studied in agents of wound infections., Conclusion: In order to reveal factors influencing on microbial etiology of mixed wound infections it is reasonable to consider the nature of interspecies interactions of causative agents.

Aim: To study genetic diversity of Pseudomonas aeruginosa strains circulating in intensive care unit (ICU), to determine the source of these strains and duration of circulation of epidemically-significant clone in the hospital., Materials and Methods: Genotyping of 106 P. aeruginosa strains isolated from patients, clinical specimens and fomites was performed by random amplified polymorphic DNA analysis with oligonucleotide primer Sh1 of 10 bp long., Results: Out of 106 P. aeruginosa isolates, 72.6% belonged to the same genotype, which was dominated in ICU during whole study period. It was established that 58.3% of examined patients were colonized by identical strains belonged to prevalent genotype that indicates the intrahospital transmission of epidemic strain., Conclusion: Obtained data show that during the period of observation (15 months) one clone of P. aeruginosa dominated in ICU, which was characterized by multiple resistance to antibiotics and caused nosocomial infection in 58.3% of patients. This confirms the need of continuous molecular-microbiological monitoring of hospital microflora in order to early detect potentially dangerous epidemic hospital strains, which are able to cause nosocomial infections.

The paper presents the results of the first isolation of the new influenza virus in Moscow and the Russian Federation, which was similar to the swine A/IIV-Moscow/01/2009(H1N1)swl strain isolated on May 24, 2009 from a Russian arrived in Moscow from the USA on May 19, 2009. The antigenic, biological, and molecular genetic properties of this virus were studied. The virus was isolated on MDCK and chick embryos, the hemagglutination titers being 1:8-1:16 AE; the infectious titers being 6.51g of the tissue cytopathogenic infective dose (TCID50) and 7.01g of the common infective dose (CID50). The virus was sensitive to arbidol, ribavirin, oseltamivir, and resistant to rimantadine. The complete virus genome was sequenced; the data were accepted to the Gen Bank on May 28, 2009 under GQ219584-GQ219590 and GQ202724. The significant gene substitution of neuraminidase Asp for Gly in position 451, which has been undetectable in any other strain published in the Gen Bank by the present time is unique only to A/IIV-Moscow/01/2009 (H1N1)swl. The virus has been deposited in the State Collection of Viruses, D. I. Ivanovsky Institute of Virology, Russian Academy of Medical Sciences, under No. 2452 dated May 24, 2009.

Aim: To study the rate of infection of ticks captured one of the Moscow park terrains with bacteria (agents of tick borreliosis and anaplasmosis)., Materials and Methods: Rates of infection of dried ticks with agents of main tick-borne bacterial infections (tick borreliosis and anaplasmosis) were determined by nested PCR., Results: In May-June 2006, 76 ticks (40 adult females, 36 adult, males) belonged to Ixodes ricinus species were captured by the method "on flag". Number of ticks on the chosen terrain was 1.77 ticks per km2. 22.4% of ticks (12 females and 5 males) were positive for the agent of tick borreliosis--spirochete Borrelia burgdorferi sensu lato which is pathogenic for humans. The main detected pathogen was Euro-Asian genovariant of B. garinii--7 female and 5 male ticks (70.6% from total number of infected ticks) were infected with it. Five female ticks were infected with genovariant of B. afzelii. One female tick (1.2%) was infected with B. valaisiana., Conclusion: Anaplasma A. phagocytophilum causing human granulocytic anaplasmosis was not detected in captured adult ticks.

Aim: The objective of this study was to assess dynamics of prevalence of hemotransmissive viral infections (HTVI) markers as mono- or mixed infections in patients admitted to Sklifosovsky Research Institute of Emergency Medicine., Materials and Methods: Two hundred thirty-five thousands and seventy patients admitted to the Institute during 1998 - 2007 were screened for the presence of laboratory markers of HTVI (antibodies to HIV, HBsAg, and anti-HCV)., Results: The long-term screening revealed 27,942 (11.9%) persons positive for laboratory markers, of which 3,099 (11.1%) were positive for >1 marker. It was established that HIV infection as well as viral hepatitis B and C have different level of similarity and ability to coexist. HIV infection was more frequently detected in association with other infections, whereas hepatitis B and C--as monoinfections., Conclusion: Attempt to consider the mutual influence of HTVI on their prevalence from the view of modern concept of microorganisms' ecology was made. Computation of Jaccard similarity coefficient showed that HIV and HCV have the greatest abilities for coexistence, whereas HIV and HBV--the smallest.

Aim: To perform advanced antigenic characterization of meningococci belonging to serogroups A and B and circulating in Moscow according to modern nomenclature of Neisseria meningitidis strains., Materials and Methods: Method of typing of "VR" fragment of FetA protein together with methods of genetic subtyping and multilocus sequence typing was used., Results: Detailed information about studied strains was inputed in Internet database--http://pubmlst.org/neisseria/. Typing of serogroup B strains did not allow to define dominating variant of "VR, fragment of FetA protein which is in accordance with subtyping data obtained previously. Serogroup A strains were notable for less variability of "VR" fragment variants: 6 variants were detected. For the majority of serogroup A strains, it was possible to trace connection between belonging of the strain to particular genetic subgroup and its revealed antigenic profile. For strains from genetic subgroup VI, antigenic profile P1.5-2, 10; F1-5 detected in 14(18%) strains was typical, whereas antigenic profile P1.5-2, 10; F3-5 was typical for genetic subgroup X and was detected in 50 (63%) strains. Antigenic profile P1.5-2, 10-67; F3-5 was detected in 5 (6%) strains, and other 10 antigenic profiles were revealed in one strain each., Conclusion: Prevalence of strains with antigenic profile P1.5-2, 10; F3-5 is explained by change of predominant genetic subgroup from subgroup VI to subgroup X in Moscow population serogroup A meningococci observed after 2003.

Testing of immunocompromised patients for markers of beta-herpesviruses--human herpesvirus type 6 (HHV-6) and cytomegalovirus (CMV), as well as gamma-herpesvirus--Epstein-Barr virus (EBV), revealed that all mentioned infections are frequently detected, mainlyas mixed infections. Chronic HHV-6 infection was diagnosed in more than half of the patients, whereas markers of acute phase of CMV and EBV infections were detected in 25% and 15% of patients respectively.

The paper gives the results of using ozeltamivir in patients with influenza during the epidemic upsurge of morbidity in Russia in the 2006-2007 season. Comparative analysis of nucteotide sequences of viral strains isolated from the patients taking ozeltamivir revealed no marker mutations determining the resistance to this chemical.

Human herpes virus type 6 (H HV-6) and cytomegalovirus (CMV) seroprevalence among HIV-infected and healthy women of childbearing age from Chelyabinsk and Moscow was measured with ELI - SA test-system for detection of antibodies to respective viruses. Seropositivity to HHV-6 was detected in 93.5% of HIV-infected women, that is 1.2 times more often than in healthy woman of child-bearing age. Seropositivity to CMV was detected in 87% of HIV-infected women, that is 1.05 times more often than in healthy woman of childbearing age. Low titers to HHV-6 and CMV were detected in seropositive healthywomen more frequently (in 44.1 and 48.4% respectively) whereas intermediate and high titers prevailed in HIV-infected women (in 73.9% for HHV-6 and 31.4% for CMV).

Influenza and acute respiratory viral infections (ARVI) continue to be one of the most actual medical and social-economic problem. But problem of high incidence of ARVI often regarded as problem of influenza only. Information about methods of prognosis of massive spreading of ARVI complex and universal system of their prophylaxis including influenza is presented in the article.

Pulse electrophoresis of group A streptococci (GAS) isolated in 1998 - 2004 in Moscow from patients during outbreaks of tonsillitis in child institutions, military unit and also from sporadic cases in children and adults was performed. During analysis of 48 strains 16 pulse electrotypes were recognized. These data allow to consider that in different child institutions (and other groups) accumulation and spreading of most adapted to such environment variants of GAS are take place and population structure of GAS in such institutions is unique on molecular and other characteristics. This study showed that complex typing of GAS (use of pulse electrophoresis and PCR for detection of erythrogenic toxins A and C) will help to improve molecular-epidemiologic surveillance for streptococcal group A infection, development of recommendations on reduction of morbidity from this infection and also to decrease risk of its severe forms, complications and mortality.

A total of 167 children with bronchial asthma (BA) have been examined for the mycoplasma infection rate. Among investigated patients 62,8% were infected with one or more mycoplasma species. The prolonged persistence in patient body as well as biological properties of mycoplasmas give grounds to consider these agents as a risk factor in the development of the allergy-infection-borne BA and its relapses.

The analysis of meningococcal strains of different serogroups, isolated from the liquor of patients in Moscow, which was carried out with the method of multilocus sequencing-typing (MLST), was presented. At the periods of epidemic morbidity rises in Moscow the prevalence of group A meningococcal strains, belonging to subgroups III with sequence-types 5 (in the 1970s) and 7 (in 1996), was noted, and at a period between epidemics strains of genetic subgroups VI and X were isolated. Meningococcal strains, groups B and C, isolated in 1995 - 2002, had, as a rule, unique sequence-types, differing both one from another and from N. meningitidis sequence-types detected in other countries. Among group B meningococci the prevalence of strains belonging to clonal complex ST-18 was noted, while for group C meningococcci strains belonging to clonal complex ST-41/44 were most typical. Such genetic variability of circulating meningococci was regarded as characteristic of the period between epidemics, observed in Moscow since the end of the 1980s.

The article deals with the results of the 10-year study of the synanthropic urban foci of leptospirosis on the territory of Moscow. Information on the manifestation of the activity of the foci under study, rodents serving as the reservoir of infection in these foci, the etiological structure of the leptospires among these rodents, the state of leptospirosis morbidity among humans is presented.

The characterization of the pulse-electrotypes of L. monocytogenes, isolated in 2003-2004 in Moscow from different sources, is presented. Among the cultures, isolated from humans, one outbreak pulse electrotype was detected and from different objects in buildings where a wide variety of food products was produced several probably related and unrelated pulse-electrotypes were obtained. The conclusion was made that several independent L. monocytogenes clones existed on the territory of Moscow, and many products supplied to retail trade and public catering enterprises were contaminated with these clones. Pulse electrophoresis was shown to be the most effective method for intraspecific typing and the study of the molecular epidemiology of listeriosis. Grounds for the necessity to improve the microbiological diagnostics of L. monocytogenes infection are given.

A salmonellosis outbreak, caused by S. typhimurium, was investigated with the use of some microbiological and molecular-biological methods of typing. This investigation revealed that the outbreak was caused by the "outbreak" electrotype of the multi-resistant variant of the infective agent, found to have several plasmidovars. The possibilities and limitations of typing by sensitivity to antibiotics and plasmid DNA profile were shown. These methods of intraspecific typing were regarded as methods making it possible to establish the heterogeneity of S. typhimurium with the use of intraclonal markers.

Medical personnel is one of the main risk groups with respect to hospital infections for which hepatitis B and C are professional pathology. The degree of the risk of contamination of the personnel greatly depends on the character, urgency and duration of professional activity, accuracy in following the rules of personal safety while working with infected material, the proportion of patients contaminated with viruses of hepatitis B and C in specialized departments, the epidemiological danger of therapeutic and diagnostic methods, the possibility of reliable disinfection and sterilization of the equipment. In the system of measures aimed at preventing the infection of the medical personnel a special role belongs to the specific immunoprophylaxis of hepatitis B. Prolonged experience in the immunization of the medical personnel of a big hospital demonstrates a high immunological and epidemiological efficiency of the use of vaccine against hepatitis B, produced by the Research and Manufacturing Complex "Combiotech", Russia.

The article deals with the results of the epidemiological analysis of the spread of salmonellosis in Moscow. During the last 15 years the proportion of Salmonella enteritidis was about 80% and more. After salmonellosis morbidity decreased to 19.9 per 100,000 of the population (the minimum level) in 1985 its sharp rise was noted. Morbidity rate increased from 31.7 in 1987 to 55.1 per 100,000 in 1988, then to 93.9 per 100,000 of the population in 1989 (the maximum level). In the subsequent years up to 1996 a decrease in morbidity rate was observed, but in 1997-2002 morbidity rate stabilized within 29.8-35.7 per 100,000 of the population without a perceptible tendency towards decrease. The sharp increase of the etiological role of S. enteritidis which led to a wide spread of Salmonella infections was caused by the "chicken-egg" factor of their transmission. High morbidity rate among children and adults, registered all the year round, is the consequence of the constant epidemic activity of this factor.