[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.

[Seroprevalence of hepatitis E virus (Hepeviridae: Orthohepevirus: Orthohepevirus A) among pregnant women in the highly endemic region of Kyrgyzstan].

INTRODUCTION: Hepatitis E (HE) is an important public health problem worldwide and is especially significant for pregnant women, among whom the associated mortality rate reaches 25%. The distribution of HE serological markers in this cohort in the endemic regions of Central Asia is poorly understood. The aim of the study was to assess the seroprevalence of HEV among pregnant women in the region of Kyrgyzstan where an increased incidence of HE is reported. MATERIAL AND METHODS: Blood serum of pregnant women, obtained from medical institutions in Bishkek, city of Osh and Osh region in the period from September 2016 to October 2019, and the control group of clinically healthy women were tested using the test systems "DS-IFA-ANTI-HEV-G" and "DS-IFA-ANTI-HEV-M" (NPO "Diagnostic Systems", Russia). RESULTS: IgG antibodies to HEV were detected in 87 (5.9%) of 1472 examined pregnant women, IgM antibodies -in 64 (4.8%) of 1378, while 5 (0.34%) samples were simultaneously positive for IgG and IgM antibodies. The rates of detection of antibodies to HEV in women of three age categories from 17 to 36 years old in the studied and control groups were similar. The analysis of the seasonal dynamics of HEV seroprevalence in pregnant women in the period from February to September 2019 showed a tendency towards an increase in the values of the positivity coefficient of specific IgM antibodies by the beginning of the autumn. Antibodies to HEV were detected with highest frequency in women from Osh region. CONCLUSIONS: A high risk of HEV infection for pregnant women in the surveyed region had been shown.

[The estimation of the hepatitis E proportion in the etiological structure of acute viral hepatitis in certain regions of of Kyrgyzstan.]

Despite the fact that the Kyrgyz Republic (KR) belongs to the highly endemic regions of the world for hepatitis E, the true extent of the spread of this infection in the country remains poorly understood. It was estimated the prevalence of serological markers of hepatitis E virus (HEV) infection among patients with acute viral hepatitis (AVH) from the regions of the Kyrgyz Republic with a high level of seroprevalence previously established by us. Blood sera samples of hepatitis patients who were admitted to hospitals of Kyrgyzstan in the period 2018-2019 were examined by the enzyme immunoassay method using the kits «DS-ELISA-Anti-HEVIgG¼ and «DS-ELISA-ANTI-HEV-IgM¼ (RPC Diagnostic Systems, Russia). IgG and IgM antibodies to HEV were detected in 103 of 344 studied samples (29.9%). Most often, seropositive specimens were detected among people of age groups under 20 and over 40 years old. Hepatitis with the fecal-oral mode of transmission was dominated in the structure of AVH: the specific gravity of hepatitis E was 47.9%, hepatitis A - 35.32%. Markers of mixed infections with other hepatitis viruses have been detected in 40.4% IgM-positive individuals. Thus, high prevalence of serological markers of HEV infection in the territory of Kyrgyzstan during the interepidemic period had been shown. The necessity of including the determination of serological markers of hepatitis E into the algorithm for the comprehensive diagnosis of AVH in patients of all age groups with liver pathology had been confirmed.