Detection of circulating HCV recombinant form RF1_2k/1b in blood serum of patients by real-time RT-PCR.

Globally, about 70 million people are infected with the hepatitis C virus (HCV), and about 400 thousand people die annually from chronic hepatitis C complications. The management of patients with chronic hepatitis C may require HCV genotyping, since the efficiency of some widely used antiviral drugs strongly depend on the viral genotype and/or subtype. The most prevalent HCV circulating recombinant form, RF1_2k/1b, is misclassified as genotype 2 by many commercial HCV genotyping kits, based on the RT-PCR analysis of the 5' untranslated region of the HCV genome. This leads to inappropriate patient treatment, since the accepted treatment schemes for HCV genotype 2 are ineffective for the RF1_2k/1b. Here we describe a method for detecting the RNA HCV RF1_2k/1b in blood samples by RT-PCR analysis of two regions in HCV genome (5'UTR and NS5b). The method was tested on 240 blood serum samples from HCV infected patients, in which HCV genotype was defined as 2 or mixed (2+1 or 2+3) by the two commercial genotyping kits "OT-Hepatogen-C genotype" ("DNA-Technology", Moscow) and "RealBest RNA HCV-1/2/3" ("Vector- Best ", Novosibirsk). 50 (20.8%) RF1_2k/1b cases were revealed, including three mixed infections: RF1_2k/1b + 1a, RF1_2k/1b + 3a, RF1_2k/1b + 1b. In all cases, the accuracy of HCV typing by the proposed method was confirmed by Sanger sequencing and phylogenetic analysis. The method is easy to implement into clinical practice and may be used in clinical settings equipped for RT-PCR analysis to correctly identify the recombinant variant RF1_2k/1b.

[Clinical and laboratory parallels at pathological and borderline states associated with lipopolysaccharides.]

The review systematizes data on the structure of lipopolysaccharides (LPS) and their role in physiological and systemic pathological processes. The analysis of literature and our own data is of scientific and practical interest for specialists in the field of clinical laboratory diagnostics, anesthesiologists, resuscitators, therapists, immunologists and obstetrician-gynecologists, including studies on the role of LPS in unique three-component systems - «mother-placenta-fetus¼. The prospects of using LPS as immunomodulatory, including for the treatment of infectious diseases, are justified. It is shown that along with their use for the correction of immunodeficiency or the development of new adjuvants and vaccines, it is possible to use their high regulatory activity even at the epigenetic level. The possibility of the prophylactic and therapeutic use of LPS in the context of an alternative solution to the problem of antibiotic resistance of bacteria is discussed.

[Creation of a molecular genetic test system for early diagnosis and evaluation of the effectiveness of treatment of inflammatory periodontal diseases.]

Inflammatory periodontal diseases represent a serious dental and general medical problem due to the high prevalence among the adult population, the presence of clinical forms leading to the destruction of the dentition and tooth loss, insufficient treatment effectiveness and the frequency of relapse, including in connection with the formation of biofilms. A molecular genetic test system has been developed to evaluate the content of periodontopathogenic microorganisms Porphyromonas gingivalis, Treponema denticola, Streptococcus oralis, Streptococcus sanguis and Streptococcus sobrinus in the contents of periodontal pockets. The analytical characteristics of the test system were determined, and testing was carried out on clinical samples of patients with chronic generalized periodontitis of moderate severity. The constructed diagnostic kit allowed us to conduct a comparative analysis of the effectiveness of various types of treatment of inflammatory periodontal diseases based on quantitative data on the content of bacteria in the contents of periodontal pockets.

[Ixodic tick-borne borreliosis: clinical and laboratory features.]

For a differentiation of clinical forms of ixodic tick-borne borreliosis clinical laboratory assessment of features of the most often defined hemostasis and complete blood count (CBC) indicators is carried out. Ixodic tick-borne borreliosis in the territory of the Republic of Bashkortostan is characterized by mainly erythematous forms of a disease with a medium-weight current. At the same time some increase in quantity of platelets was noted that could be caused by irritation of a megakaryocytic sprout of bone marrow in the conditions of infectious process and have compensatory character whereas other indicators of CBC and hemostasis at this clinical form practically didn't change. Not numerous cases of the ixodic tick-borne borreliosis non-erythematous forms clinically proceeded more hard, with significantly more expressed toxicinflammatory syndrome. At the same time it was followed by significant shifts in biochemical parameters, indicators of CBC and hemostasis. The clinical laboratory features of erythematous and non-erythematous forms of ixodic tick-borne borreliosis revealed as a result of the conducted researches reflect character of a course of disease and can serve for assessment of severity, the forecast of the infection and justification of this treatment.

[The algorithms of etiologic laboratory diagnostic of parenteral viral hepatitis].

The article considers methods of laboratory diagnostic of parenteral viral hepatitis. The approaches ensuring single-valued differentiation of infected patients are determined. The various methods of evaluation of activity of infection process are presented. The algorithm of complex laboratory analysis concerning presence of parenteral viral hepatitis (B, C, D, G, TT, SEN) was proposed to ensure maximal informative minimum of laboratory analyses permitting fast and single-valued interpretation of received diagnostic data.

[THE MOLECULAR GENETIC CHARACTERISTIC OF SPECIES CONTENT OF SALIVA AND GINGIVAL RECESS UNDER PERIODONTITIS].

The examination was carried out of samplings of 110 patients with periodontitis (observation group) and 60 patients without pathology of periodont (comparison group). The polymerase chain reaction was used to analyze samples of saliva and contents of periodontal recesses for detecting species-specific DNA fragments of Porphymmonas gigngivalis, Streptococcus macacae, S. mutans, S. oralis, S. salivarius, S. sangis, S. sobrinus, Treponema denticola. In patients with periodontitis S. mutans, S. oralis S. sobrinus were reliably more often detected in the content of periodontal recesses and S. mutans, S. sobrinus i in saliva. In the observation group the rate of detection of association S. mutans--S. oralis--S. sangis--S. sobrinus was significantly exceeded (up to 15.6%, X2 = 9.1, p = 0.004). In ten days of effective treatment of periodontitis reliable decreastng of rate of detection of S. wasoralis, S. sobrinus was observed in contents of periodontal recesses but not in of saliva. The detection of S.sobrinus using technique of polymerase chain reaction in contents of periodontal recesses and/or saliva of patients with periodontitis has a diagnostic value. The detection of S.sobrinus in contents of periodontal recesses is significant both in monoculture and in association S. mutans--S. oralis--S. sangis--S.sobrinus. The absence of S. sobrinus in contents of periodontal recesses testifies effectiveness of treatment of main disease (periodontitis).

[The epidemiologically valid comparative evaluation of informativeness of techniques of laboratory diagnostic of zooanthroponosic trichophytosis.]

The analysis of long standing morbidity of zooanthroponosis trichophytosis was carried out. The comparative laboratory analysis (microscopy, mycologic technique, polymerase chain reaction) of clinical samples from 111 patients (aged 7-14 years) with clinical diagnosis of trichophytosis (T.verrucosum), 110 patients (aged 3-14 years) with clinical diagnosis of trichophytosis (T.mentagrophytes) and 186 patients with other dermatological diseases (psoriasis, eczema) (negative control). It is demonstrated that in evaluation of diagnostic effectiveness of laboratory methods in case of mycosis induced by T.verrucosum and T.mentagrophytes the optimal choice is the examination of groups of children aged 7-14 and 3-14 years correspondingly. Under trichophytosis (T.verrucosum) sensitivity of polymerase chain reaction amounted to 98.2 (96.5-99.9)%; p<0.05, specificity - 97.5 (95.1-99.9)%; p<0.05 and diagnostic effectiveness - 97.9 (95.9-99.9)5; p<0.05. Under trichophytosis (T.mentagrophytes) sensitivity of polymerase chain reaction corresponded to 97.3 (94.8-99.8)%; p<0.05, specificity - 97.1 (94.6-99.6)%; p<0.05 and diagnostic effectiveness - 97.2 (95-99.4)5; p<0.05.