Development and Validation of a Protein Array for Detection of Antibodies against the Tick-Borne Pathogen Borrelia miyamotoi.

Current serological tests for the emerging tick-borne pathogen Borrelia miyamotoi lack diagnostic accuracy. To improve serodiagnosis, we investigated a protein array simultaneously screening for IgM and IgG reactivity against multiple recombinant B. miyamotoi antigens.

[The information content and predictive value of cardiac markers in myocardial infarction in the elderly.].

Myocardial infarction in old age is the leading pathology and cause of deaths. The aim of the study was to evaluate the prognostic significance and informative value of cardiac markers of blood in elderly patients with myocardial infarction. The study of cardiac markers in 107 patients with myocardial infarction at the age of 65-75 years and in 83 patients with myocardial infarction at the age of 50-60 years was carried out by immunochromatographic, bioluminescent methods and by electrophoresis on cellulose acetate, followed by the calculation of informativeness and prognostic coefficients.

Whole genome sequencing of Borrelia miyamotoi isolate Izh-4: reference for a complex bacterial genome.

Background: The genus Borrelia comprises spirochaetal bacteria maintained in natural transmission cycles by tick vectors and vertebrate reservoir hosts. The main groups are represented by a species complex including the causative agents of Lyme borreliosis and relapsing fever group Borrelia. Borrelia miyamotoi belongs to the relapsing fever group of spirochetes and forms distinct populations in North America, Asia, and Europe.

Evaluation of the population heterogeneity of TBEV laboratory variants using high-throughput sequencing.

We studied minor variants within two tick-borne encephalitis virus (TBEV) populations with a common ancestor: the mouse brain-adapted variant EK-328c and the tick-adapted variant M. High-throughput sequencing with custom amplicons from RT-PCR viral RNA was performed on Illumina MiSeq 2*250 paired-end v2 chemistry. Using the LowFreq program (default settings) and Sanger-sequenced consensus as a reference, variants with an abundance of 1 % and above within the studied populations were identified.

Whole-Genome Sequencing of Six Clinical Strains Isolated in Russia.

Here, we report the whole-genome sequence of six clinical isolates from the Russian Federation. Using two independent next-generation sequencing platforms, we determined the complete sequence of the chromosome and several plasmids. All strains have an Asian genotype with 99.

Enrichment of Viral Nucleic Acids by Solution Hybrid Selection with Genus Specific Oligonucleotides.

Despite recent advances, our knowledge of potential and rare human pathogens is far from exhaustive. Current molecular diagnostic tools mainly rely on the specific amplification of marker sequences and may overlook infections caused by unknown and rare pathogens. Using high-throughput sequencing (HTS) can solve this problem; but, due to the extremely low fraction of pathogen genetic material in clinical samples, its application is only cost-effective in special, rather than routine, cases.

Specificity of SNP detection with molecular beacons is improved by stem and loop separation with spacers.

Molecular beacons (MBs) are valuable tools in molecular biology, clinical diagnostics and analytical chemistry. Here we describe a novel approach for the design of MBs with nucleotide or non-nucleotide linkers between the stem and loop regions. Such modified MBs have significantly improved specificity and performance for single nucleotide polymorphism (SNP) detection.

Complete Genome Sequence of Human Adenovirus 7 Associated with Fatal Adult Pneumonia.

Human adenovirus 7 (hAdv7) 19BOVLB/Volgograd/Rus/2014 was isolated from the autopsy material from an adult with fatal pneumonia in Volgograd, Russia, in March 2014. Whole-genome sequencing of the virus isolate was performed.

[GENETIC CHARACTERISTICS OF INFLUENZA A/H3N2 AND B VIRUSES THAT HAD CIRCULATED IN RUSSIA IN 2013 - 2015].

Aim: Establish genetic characteristics, carry out phylogenetic analysis and determination of molecular markers of resistance to etiotropic preparations against influenza A/H3N2 and B viruses that had circulated in Russia in 2013 - 2015.

Materials And Methods: 80 biological samples containing influenza A/H3N2 virus RNA and 31 samples containing influenza B virus RNA were studied. Sequencing of PCR fragments was carried out inABI-3 100 PRIZMTM GeneticAnalyzer (AppliedBiosystems, USA) and using MiSeq (Illumina, USA).

Development and evaluation of the RT-PCR kit for the rabies virus diagnosis.

To improve the diagnosis, surveillance, and control for the rabies virus, a kit for hybridization-triggered fluorescence detection of rabies virus DNA by the RT-PCR technique was developed and evaluated. The analytical sensitivity of the kit was 4*10 GE per ml. High specificity of the kit was shown using representative sampling of viral, bacterial, and human nucleic acids.

Whole-Genome Sequence of Mycobacterium bovis BCG-1 (Russia).

BCG vaccine (Mycobacterium bovis BCG-1 [Russia]) is the most important component of tuberculosis prophylaxis in Russia. This study represents the complete genome sequence and genetic characteristics of M. bovis BCG-1 (Russia), which has been used to manufacture BCG vaccine in Russia and in some other countries.

Sequencing of the complete genome of an araphid pennate diatom Synedra acus subsp. radians from Lake Baikal.

High-throughput method of sequencing was applied to determine the complete nucleotide sequence of an araphid pennate diatom Synedra acus subsp. radians from Lake Baikal (East Siberia). The assembled genome has a total length of 98 Mbp, the mean coverage is 33x.

[The development of reagents set in the format of DNA-chip for genetic typing of strains of Vibrio cholerae].

The necessity of development of methods of genic diagnostic of cholera is conditioned by continuation of the Seventh pandemic of cholera, taxonomic variability of strains of Vibrio cholerae involved into pandemic and also permanent danger of delivery of disease to the territory of the Russian Federation. The methods of genic diagnostic of cholera make it possible in a comparatively short time to maximally minutely characterize strains isolated from patients or their environment. The article presents information about working out reagents set for genetic typing of agents of cholera using DNA-chip.

Prevalence of Kemerovo virus in ixodid ticks from the Russian Federation.

The prevalence of Kemerovo virus in ixodid ticks collected in 2008-2012 from 11 regions of the Russian Federation was investigated by real-time reverse-transcription PCR (RT-PCR). The presence of Kemerovo virus in Ixodes persulcatus, Ixodes ricinus, and Dermacentor reticulatus was confirmed. Virus prevalence depended on the region and varied from zero to 10.

Draft Genome Sequencing of Vibrio cholerae O1 El Tor Isolates Collected in the Russian Federation from Imported Cholera Cases.

We report the draft genome sequencing of five Vibrio cholerae O1 El Tor clinical isolates collected in the Russian Federation from imported cholera cases in 2006, 2010, and 2012. In the initial phylogenetic analysis, one isolate clustered with the Haiti/Nepal-4 group.

[The new possibilities in serologic diagnostic of Ixodes mite-borne borreliosis using ImmunoChip].

The new kit of reagents in format of the immunochip "ImmunoChip Borreliosis" for multiplex serologic analysis of mite-borne borreliosis demonstrated high diagnostic sensitivity and specificity. The percentage of detection of specific immunoglobulins was higher in "ImmunoChip Borreliosis" as compared with screening results in immune enzyme analysis. The high correlation between results of testing in immunochip and data of immune blotting is demonstrated to.

[Phylogenetic analysis of genomes of Vibrio cholerae strains isolated on the territory of Rostov region].

Aim: Determination of origin of 2 Vibrio cholerae strains isolated on the territory of Rostov region by using full genome sequencing data.

Materials And Methods: Toxigenic strain 2011 EL- 301 V. cholerae 01 El Tor Inaba No.

[The detection of activating somatic mutations in gene KRAS using pyrosequencing technique].

The technique to detect all possible variants of mutations in 12, 13 and 15 codons of gene KRAS was developed on the basis of the pyrosequencing technology. The analytical characteristics of the developed technique were identified. The limit of detection for mutations G34T, G35A and G38A detected on the cloned control samples consisted 3%.

Draft Genome Sequences of Vibrio cholerae O1 ElTor Strains 2011EL-301 and P-18785, Isolated in Russia.

We report the draft whole-genome sequences of two Vibrio cholerae O1 strains, the environmental toxigenic strain 2011EL-301 and the clinical nontoxigenic strain P-18785, both isolated in Russia. Some basic data comparing the two against the GenBank repository are provided.

[Downregulation of human CCR5 receptor gene expression using artificial microRNAs].

Chemokine receptor CCR5 is essential for human immunodeficiency virus (HIV) entry into the sensitive cells. The CCR5 inactivation is believed to be one of the promising approaches in HIV therapy, including gene therapy. A powerful mechanism that enables to regulate gene expression is RNA interference which could be exploited to knockdown CCR5 gene.

[The verification possibilities of immunochips under low content of antibodies to Core-antigen of virus of hepatitis C].

The detection of antibodies to Core-antigen of virus of hepatitis C in test-systems for solid-phase immune-enzyme analysis with low optical density can be a result not only of true availability of antibodies but an effect of nonspecific reaction of blood serum. The diagnostic possibilities of immunochips to be used in immune-enzyme analysis for verification of availability of markers of viral hepatitis C were investigated in conditions of low positive reaction of blood serum to core-antigen. It is established that immunochips and immunoblots have similar specificity concerning detection of antibodies to Core-antigen.

[Treatment of HIV-infection by means of gene therapy].

Current methods of HIV treatment can contain a progression of the disease; however they do not lead to a cure. Lifelong antiretroviral therapy is therefore necessary, leading to problems of cost and toxicity of chemical drugs. The recent advances in science have allowed a new approach to the HIV-treatment - gene therapy.

[The detection of nifedipine in biological materials].

The objective of the present study was to find optimal conditions for the isolation of nifedipine from biological materials by ethylacetate. It was shown that nifedipine can be purified from co-extracted substances of the biological material on a Silasorb C-18 column with the size of the particles 30 microns. The authors propose to use thin-layer chromatography, IR spectrophotometry, and reverse-phase high performance liquid chromatography for the identification and quantitative determination of nifedipine extracted from cadaveric liver samples.

[Design of an immunochip for separate detection of hepatitis C virus antibodies].

A test kit as an immunochip designed for the diagnosis of hepatic C virus (HCV) has a high sensitivity and specificity. Recombinant HCV antigens were separately immobilized on the activated slides together with internal controls. Serum test results were red by ScanArray Express.