[Line immunoassay for detection of IgG antibodies to hepatitis E virus (Hepeviridae, Orthohepevirus, Orthohepevirus A)].

INTRODUCTION: The diagnostic efficacy of methods for hepatitis E serodiagnostic varies over a wide range; therefore, the combined use of tests of various formats is recommended. The aim of the research was to develop a test system for the detection of IgG antibodies to hepatitis E virus (HEV) in human serum by linear immunoassay (LIA). MATERIAL AND METHODS: Serum samples from patients with hepatitis and healthy individuals were tested using commercial enzyme-linked immunosorbent assay systems for the presence of IgG antibodies to viral agents causing hepatitis and other infections associated with liver pathology. Recombinant antigens ORF2 and ORF3 of HEV genotypes 1 and 3 were used. The "RecomLine HEV IgG/IgM" reagent kit (Mikrogen GmbH, Germany) was used as a comparison test system. RESULTS: The first Russian diagnostic kit "Blot-HEV", designed to detect IgG antibodies to individual HEV proteins in human serum using LIA, was developed. The antigenic base is represented by strips of a nitrocellulose membrane with immobilized recombinant antigens ORF2 (aa 406-660) and ORF3 (aa 1-113) of HEV genotypes 1 and 3, and control antigens in the form of discrete lines. The conjugate was mouse monoclonal antibodies to human class G immunoglobulins labeled with horseradish peroxidase. The chromogen solution contained the 3,3',5,5'-tetramethylbenzidine. A visual and digital recording of results was provided. The analytical sensitivity of the test kit was 0.625 IU/ml for ORF2 antigens and 2.5 IU/ml for ORF3 antigens. The absence of the influence of endogenous interfering substances on the results of the analysis and the absence of cross-reactions with antibodies to hepatitis pathogens of the other etiologies had been shown. The sensitivity of the test system compared to the "RecomLine HEV IgG/IgM" kit was 92%, specificity 97%. Shelf life in condition of storage was determined to be 12 months. CONCLUSIONS: The developed test can be used to confirm the results of ELISA in laboratory diagnosis of hepatitis E.

[Clinical significance of interleukin-28B and RNAse L gene polymorphism determination in patients with chronic viral hepatitis C].

AIM: Determination of frequency of occurrence and clinical significance ofinterleukin-28B (IL28B) and RNAse L gene polymorphism in patients with chronic hepatitis C (CHC). MATERIALS AND METHODS: 104 hospital patients with CHC (65% male; 63% with genotype 1 hepatitis C virus - HCV) were examined. 70 patients received therapy with interferon (IFN) and ribavirin (RBV). Single nucleotide polymorphism (SNP) of IL28B gene 39743165T>G (rs8099917), SNP 39738787C>T (rs12979860) and RNAse Lgene (1385G>A) were determined by polymerase chain reaction. RESULTS: The frequency of detection of "favorable," SNP allele variants of IL28B gene in patients with CHC was lower than in population of the European region. In patients with genotype 1 HCV, mutant alleles in SNP 39743165T>G (p=0.045) and 39738787C>T (p=0.005) occurred more frequently than in patients with other virus genotypes. Highervalues of alanine aminotransferase in patients with genotype CC 39738787C>T were detected. Frequencies of SNP variants of IL28B and RNAse L gene did not differ depending on the speed of disease progression (p>0.5). Response to IFN/RBV therapy was higher in "favorable" TT (SNP 39743165T>G) and CC (SNP 39738787C>T) variants. CONCLUSION: Examination for IL28B gene SNP 39738787C>T is recommended before the start of IFN/RBV therapy in all the patients with genotype 1 HCV as a prognostic factor on the therapy response. RNAse L gene SNP 1385G>A does not have a clear clinical significance in CHC.

[Antibodies to different virus antigens in patients with chronic hepatitis C].

A total of 176 hospital patients with chronic hepatitis C (CHC), among them 110 males and 66 females, were examined. The spectrum of antibodies to four hepatitis C virus (HCV) proteins (core, NS3, NS4, NS5) and in 142 patients --IgM antibodies to HCV (anti-HCV IgM) were determined. In 92% of the CHC patients antibodies to core, NS3 and NS4 proteins were simultaneously detected. Differences in the detection of antibodies to HCV in males and females were not statistically reliable. In CHC patients aged up to 20 years anti-NS4 and anti-NS5 were less frequently detected. Among males of different age groups reliable differences in the detection rate of anti-NS5 were registered, while among females of different age groups no such differences were observed. With the increase of age these antibodies were detected somewhat more often. In females over 60 years anti-HCV IgM occurred more often than in males of the same age. The levels of alanine aminotransferase (ALT) were higher in persons with the presence of anti-NS5 and anti-HCV IgM than in persons with their absence. In all groups of CHC patients with biochemical activity and liver cirrhosis the detection rate of anti-HCV IgM was significantly higher than in patients with normal ALT activity. The antibody spectrum with the simultaneous absence of HCV IgM and anti-NS5, while found to contain antibodies to other HCV antigens, was registered significantly less frequently in patients with moderate and high CHC activity and the liver cirrhosis induced by HCV infection.

[Levels of some cytokines and antibodies to hepatitis C virus in patients with chronic hepatitis C].

The comparison of the levels of some cytokines (tumor necrosis factor alpha (TNF-alpha), IL-1beta, IL-2, IL-4) in the blood serum of patients with chronic hepatitis C (CHC) having different antibody spectrum was carried out. In CHC patients increased levels of the serum cytokines IL-1beta, TNF-alpha under study in comparison with cytokine levels in donor sera was noted. In patients with detected antiNS5 and antiHCV IgM and antiNS5 HCV the level of IL-1beta was significantly higher than that in CHC patients without antibodies in sera. A change in the levels of proinflammatory and anti-inflammatory cytokines in the blood sera of CHC patients may be of significant diagnostic and prognostic importance.

[Phenotypical and molecular-genetic typing of salmonellae: reality and prospects].

The importance of the intraspecific typing of salmonellae for effective epidemiological analysis is shown. The methods (phenotypical and genotypical), used for the epidemiological marking of salmonellae, are characterized, their advantages and drawbacks, as well as prospects of their further use, are given. The importance of the standardization of molecular-genetic methods to ensure data exchange between different laboratories and the investigation of international outbreaks of enteric infections is pointed out. The international projects PulseNet and Salmgene of the surveillance of Salmonella infection, based on genotyping of Salmonella isolates, are presented.

[Adhesiveness and antibiotic resistance of the causative agents of salmonella infection in Gomel' region]. / Adhezivnost' i antibiotikorezistentnost' vozbuditelei sal'monelezov v Lomel'skoi oblasti.

The adhesive capacity, resistance to antibiotics and biological properties of Salmonella strains of different serogroups, circulating in the Gomel region, were under study. Resistance to antibiotics and changes in biological properties were accompanied by an increased adhesiveness of these strains. A high degree of adhesiveness was noted in S. hadar, a new for Belarus strains.

[Autopsy-based data on radionuclide concentrations in selected endocrine organs (Vitebsk region, Belarus)]. / Soderzhanie radionuklidov v nekotorykh éndokrinnykh organakh po materialu autopsii v vitebskoi oblasti.

Gamma spectometry was used to compare 137Cs levels in certain endocrine organs of victims of general and oncological diseases of different localizations. The radionuclide distribution in the thyroid, adrenals and pancreas was studied. Elevated concentrations were found in the endocrine system of cancer patients.

[Distribution of Cs-137 in the human body with malignant neoplasms of various localizations]. / Raspredelenie 137-Cs v organizme cheloveka pri zlokachestvennykh novoobrazovaniiakh razlichnykh lokalizatsii.

137Cs-concentrations and distribution in internal organs and malignant tumor tissues from cadavers and patients operated on for cancer were identified by gamma spectrometry in clinics of Vitebsk Region. Non-uniform distribution patterns in different organs were registered, irrespective of the presence or absence of malignancy.

[Distribution of Cs-137 in bodies of inhabitants of the Vitebsk Region]. / Raspredelenie 137Cs v organizme cheloveka na primere zhitelei Vitebskoi oblasti.

The content of 137Cs in the whole human body and certain internal organs has been determined using gamma-spectrometry method for inhabitants of Vitebsk Region. The average activity of 137Cs in the whole human body was obtained to be 0.038 mkCu. The unevenness of radiocesium accumulation in the investigated internal organs has been noted. The average annual doses from 137Cs for some human organs have been calculated.

[Viral hepatitis delta in the republic of Belarus]. / Virusnyi gepatit del'ta v Respublike Belarus'.

26,740 blood donors and persons of high risk groups with respect to HBV infection, residing in different regions of Belarus, were examined for the presence of HBsAg in 1983-1997. Of these, 1372 persons (5.1%) were found to have HBsAg, and out of 1081 HBsAg-positive persons anti-HDV antibodies (Ab) were detected in 96 persons (8.9%). In spite of a decrease in acute virus hepatitis B morbidity and in HBsAg carriership, the occurrence of anti-HBV Ab remained stable during the period of 16 years and was equal, on the average, about 4% among asymptomatic HBsAg carriers. Patients having tuberculosis, rheumatoid arthritis, diabetes mellitus, hematological diseases, chronic hepatitides and cirrhosis of the liver were an important reservoir of HBV and HDV infections for regions with the low level of the spread of HBV. A decrease in the detection rate of anti-HDV Ab in patients with cirrhosis of the liver from 47.6% to 15.4% was noted. In 1991-1997 a decrease in the detection rate of anti-HDV Ab in patients with chronic hepatic lesions in comparison with 1983-1990 was observed, and in the age group older than 50 years this decrease was from 33.3% to 8.3%. This difference was particularly pronounces in patients with cirrhosis of the liver: 53.9% and 7.7% respectively.

[Radionuclide pollution of the Vitebsk region]. / Zagriaznenie territorii Vitebskoi oblasti radionuklidami.

The 137Cs content has been determined for probes of flora, fauna, food, soil and autopsy material from Vitebsk Region. The 137Cs content was established not to be more, than the Republic Control Levels of 1992. The dynamic of 137Cs accumulation in potato for 1981-1995 has been shown. The presence of 137Cs was connected with radionuclide air transferring from radiation contaminated areas from nuclear power objects and from "trace" of Chernobyl Accident.

[The use of wild animals in radioecological monitoring]. / Opyt ispol'zovaniia dikikh zhivotnykh v radioékologicheskom monitoringe.

Radionuclide content in muscular and osseous tissues of elks and wild boars was determined by means of spectrum analysis. The radioactive caesium accumulation dynamics in muscular tissues of elks and wild boars from 1986 till 1992 years is presented.

[The determination of the concentration of the carcinoembryonic and meconial antigens in the gastric juice of patients with stomach pathology]. / Opredelenie kontsentratsii rakovo-émbrional'nogo i mekonial'nogo antigenov v zheludochnom soke bol'nykh s patologiei zheludka.

Beta-I-MA and CEA concentration in gastric juice was studied in 74 patients with gastric pathology and healthy subjects. Elevated concentrations of beta-I-MA and CEA were found more often in the gastric juice of cancer patients and less often in that of patients without malignant pathology. No significant differences in the concentration of each antigen were found between the "cancer" and "polyp" groups. The combined use of the two test procedures was shown to sufficiently complement the CEA test. The difference in the probability of high concentrations of both antigens in gastric juice in cancer and low ones in other pathologies was highly significant (chi 2 = 15.3, p. 0.00015).

[Hepatitis B and delta infections in patients with tuberculosis]. / Gepatit B i del'ta-infektsii u bol'nykh tuberkulezom.

Clinical variants of the course and outcomes of HBV and delta infections have been studied in pulmonary tuberculosis patients in the general structure of 1465 patients who were treated for hepatitis B. Enzyme immunoassay and radioimmunoassays were used to study the detection rate of contamination markers HBV activity and delta-virus replication in 437 pulmonary tuberculosis patients. Viruses were found to be predisposed to prolonged persistence and wide spread among tuberculosis patients.

[Detection rate of antibodies against delta virus among HBsAg carriers in a region with moderate prevalence of hepatitis B]. / Chastota vyiavleniia antitel k virusu del'ta sredi nositelei HBsAg iz razlichnykh grupp naseleniia regiona so srednim urovnem rasprostraneniia gepatita B.

Radioimmunoassay and enzyme-immunoassay were used to study the detection rates of HBsAg and anti-Delta in blood donors and subjects at high risk of infection with hepatitis B (HB) in the Byelorussian SSR. The republic was found to belong to the region with a moderate prevalence of HB and low prevalence of delta-virus infection. The detection rate of anti-delta antibodies in individual population groups depends on the level of HBsAg carrier state and is closely associated with prolonged treatment in medical institutions. Patients with tuberculosis, rheumatoid arthritis, diabetes mellitus, and cirrhosis of the liver are important reservoirs of HB and delta-virus infection for a region with moderate prevalence of HB virus and low prevalence of delta-virus. Infection with delta-virus is very important in unfavourable outcomes of acute and chronic viral hepatitides in the region.

[The development and a clinical trial of an erythrocytic diagnostic agent for determining antibodies to the liver-specific lipoprotein]. / Razrabotka i klinicheskoe ispytanie éritrotsitarnogo diagnostikuma dlia opredeleniia antitel k spetsificheskomu lipoproteinu pecheni.

A simple method for the preparation of purified liver-specific protein has been developed and a sensitive and specific test-system has been created on its basis for the detection of antiLSP. The titres of antiLSP correlate with the activity of the pathological processes in patients with acute and chronic hepatitides. Immunosuppressive therapy results in elimination of antiLSP.

[HBe antigen and its antibodies in HBsAg carriers in various regions of the USSR]. / HBe-antigen i antitela k nemu u nositelei HBsAg v razlichnykh raionakh SSSR.

A total of 300 blood serum samples, containing HBsAg and obtained from donors in three regions of the USSR (the RSFSR, the Uzbek SSR and the Moldavian SSR) differing in the level of HBsAg carriership, were studied for the presence of HBeAg and antibodies to this antigen in the passive hemagglutination (PHA) test and in the enzyme immunoassay (EIA). The occurrence of HBeAg was found to depend on the level of HBsAg carriership in the region. Thus, according to the EIA results, in Gorky, Kishinev and Tashkent HBeAg was detected, respectively, in 5.5%, 12.3% and 13.3% of serum samples, the level of HBsAg carriership in these cities being, according to the results of the PHA test, 1.4%, 5.0% and 9.0%. As shown by the results of EIA, the occurrence of HBeAg increased with the rise of the titer of HBsAg, while regarding the occurrence of antibodies to HBeAg the reverse relationship was observed.