Vol 3, No 1 (2013)

Abstract. Mycobacteriosis is an infectious disease of animals and humans caused by non-tuberculosis mycobacteria including M. avium complex. Despite the fact that the transmission of M. avium from human to human has not been proved, and mycobacteriosis has been sporadic, the number of cases of disseminated forms of disease caused by M. avium among HIV-positive patients during the last ten years was increasing. Limited knowledge about the structure of M. avium population in Russia and the lack of simple methods for the microbiological diagnosis make difficult the epidemiological monitoring of mycobacteriosis. This facilitates the use of modern, efficient molecular genetic methods for the species and subspecies identification and typing of M. avium. Thus, the detection of mobile element IS901, restriction fragment polymorphism analysis of hsp65 gene and IS1245 allow the detection and subspecies identification of M. avium. The study of genomic polymorphisms of bacterial strains for the assessment of M. avium population structure became feasible due to a complex of molecular techniques: VNTR-typing, IS1245-and IS1311-RFLP-typing.

Abstract. Overall 178 Mycobacterium tuberculosis isolates recovered in 2009–2011 from newly diagnosed epidemiologically unlinked to TB patients in the Ural region of Russia have been studied. The absolute concentration method was used for drug susceptibility testing. Mutations katG, inhA and rpoB associated with resistance to isoniazid and rifampicin were detected by microchip technology («TB-Biochip»). The isolates were genotyped by real-time PCR for the detection of Beijing/non-Beijing genotypes and 15-locus MIRU-VNTR typing according to «MIRU-VNTRplus» (http://www.miru-vntrplus.org). More than half (55.1%) of 178 isolates belonged to the Beijing family, 58.7% of them were multiple drug resistant (MDR) mostly due to rpoBSer531→Leu and katGSer315→Thr1 substitutions. Fifty VNTR profiles were found in 98 Beijing isolates; 57 of them grouped into 9 clusters. The largest VNTR cluster included 23 (23.5%) Beijing isolates and 21 of them were MDR. The 80 non-Beijing isolates showed 64 distinct VNTR patterns which belonged to 6 genetic families: LAM, Ural, Haarlem, etc. Among LAM and Ural isolates 30.4% and 28.6% were MDR, respectively. The 5 of 7 MDR LAM isolates had specific mutation profile:  rpoBAsp516→Val substitution and mutations katGSer315→Thr1 and inhA_T15. The MDR Ural isolates showed the heterogeneity of mutations in rpoB gene compared to other genotypes. Taken together, these findings suggest the emergence and spread of MDR-TB in the Ural region which is characterized by circulation of MDR strains of different genotypes with the Beijing family genotype to be predominant.

Abstract. The study was performed in the Perm Krai Clinical Hospital N 2 “Heart Institute”. The aim of this study was to evaluate the antibiotic susceptibility in isolates from patients with diabetes mellitus and HIV-infected patients. In all groups involved in the study (patients with diabetes mellitus, patients with HIV infection, and control group) the predomination of the Gram-positive species have been revealed. This fact is typical for surgical departments. More isolates of Staphylococcus aureus were found in the group of HIV-infected patients compared to patients in the control group (20.6% vs 4.6%, p = 0.02, respectively). Oxacillin resistance rate in Staphylococcus aureus was higher in the group of HIV-infected patients than in patients from the control group. Significant differences in Staphylococcus epidermidis resistance rate between HIV-infected patients and patients in the control group were detected for oxacillin (60% vs 10%, p = 0.03, respectively). It is plausible that in the group of immunocompromised patients vancomycin might be recommended for empiric therapy of infections caused by gram-positive species. The Enterococcus faecalis was more often isolated from HIV-infected patients in compare with patients from the control group (34.5% vs 13.8%, p = 0.02), probably due to severe immunodeficiency in the late stages of HIV infection. The absence of significant differences in resistance rate in Staphylococcus aureus and Staphilococcus epidermidis between patients with diabetes and patients in the control group could be considered as important. This fact might be associated with judicious control of glycemia during the pre-, intra-, and postoperative time periods. Postoperative pyoinflammatory diseases remain serious obstacle for the progress in cardiosurgery. Consequently, it’s important to know the characteristics of the microflora, colonizing the patients.

Abstract. Pathogenesis of chronic hepatitis C (CHC) remains to be determined. Mechanisms of liver parenchyma damage in patients with CHC are complex and different. Cytokines play the role of intermediaries in the process of fibrosis development and chronic inflammation. In the present study levels of 27 cytokines in the blood plasma of 14 patients with CHC were tested using multiplex analysis. The liver biopsy was performed in all patients to define the activity of inflammation (histological activity index) and the degree of fibrosis. Nineteen samples of blood plasma obtained from healthy individuals were served as a control group in this study. The following cytokines were measured: IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, eotaxin, FGF-2, G-CSF, GM-CSF, IFNγ, IP-10, MCP-1, MIP-1α, MIP-1β, RANTES, PDGF-BB, TNFα and VEGF. In patients with CHC elevated levels of plasma IL-1ra, IL-6, IL-7, IFNγ, IL-12 (p70), IL-4, IL-9, IL-8, IP-10, eotaxin, MCP-1, MIP-1β, TNFα, G-CSF and GM-CSF were found in compare with the control group. At the same time levels of FGF-2 and PDGF-BB were reduced in patients with CHC in compare with controls. Differences in the production of IL-1ra, IL-6, IL-7, IFNγ, IL-12 (p70), IL-4, IL-9, IL-8, IP-10, eotaxin, MCP-1, MIP-1β, TNFα, G-CSF and GM-CSF were depend on the genotype of HCV (3a or 1b), histological activity index in liver tissue and the degree of liver fibrosis. The revealed changes of cytokine production in patients with CHC characterize different orientation of regulatory violations confirming that CHC is an immunopathological process.

Abstract. The etiology of seasonal increasing of enteroviral infection incidence in Arkhangelsk oblast in 2008–2011 has been studied. The ECHO 6, ECHO 9 and ECHO 30 viruses were isolated and identified from patients with enteroviral infection in this period. The highest enteroviral infection incidence rate in Arkhangelsk oblast which was 5,1 times higher in compare with average country index was registered in 2008. It was proved by the virological and molecular biological methods that the main etiological agent caused enteroviral meningitis cases in the seasonal outbreak in 2008 was ECHO 30 virus which was very similar to the ECHO 30 strain identified in the Veliky Novgorod in 2008. In 2009 from patients in enteroviral meningitis epidemic focus the ECHO 9 viruses were isolated. These viruses were similar to the ECHO 9 strains circulated in Russia in 2009. It was established that in the period of seasonal increasing of enteroviral infection in Arkhangelsk oblast in 2010–2011 in general circulated ECHO 6 viruses which clustered in 3 separate phylogenetic groups. Viruses ECHO 6 detected in Arkhangelsk oblast in 2011 were different from viruses identified in 2010. Results of the study confirm that monitoring of viruses in case of enteroviral infection remains the one of the important types of additional measures in the frame of Global Program of poliomyelitis eradication. This monitoring is absolutely necessary to define characteristics of epidemic process of enteroviral infection.

Abstract. The cytological evidence of cervical squamous epithelium lesions associated with HPV infection was identified as the presence of koilocytes in 16% of patients and atypical multicore cells with core and/or cytoplasm dystrophy of para-basal layer cells in 46%. The high grade squamous intraepithelial lesions were associated with the presence of HPV 16 mostly in combination with other high risk HPV genotypes.

Abstract. The new cytometric method for laboratory diagnosis of brucellosis in vitro conditions based on detection of hypersensitivity to Brucella has been developed. This test allows to differentiate vaccinal and infectious processes in case of chronic brucellosis, and to measure the level of patient sensibilization to Brucella. Thus, the test might be used to estimate intensity of post-vaccination immunity before re-immunization against brucellosis. Using of flow cytometry in the test excludes additional antigenic influence on human organism and allows to provide testing within 1 hour.