Approbation of the experimental series of latex agglutination and immune chromatography test-systems was carried out for identification of Bacillus anthracis spores and vegetative cells respectively. These test-systems are suitable for rapid diagnostics of anthrax and have a number of advantages over already-existing commercial tests.
Tick-borne borreliosis (Lyme disease-LD) is caused by pathogenic Borrelia spirochetes that is transmitted through bite of Ixodes ticks to humans and animals. In the Russian Federation, borreliosis registered with an index of 6-7 per 100,000 people annually. In reality, LD morbidity in Russia is much higher because Russian strains develop less erythematous rashes compared to North American strains, thus missed by physicians in most of the early cases, and current serology tests have insufficient sensitivity as well. The aim of this work was to improve the sensitivity and specificity of serology tests for LD in Russia using rationale-designed Borrelia antigens. It was anticipated that sensitivity of LD sero-diagnosis will be higher if antigen for test-systems are derived from a strain that is circulated in a geographical region of test application. A large portion of the Russian population lives in the Central region. Thus, effort has been made to create a serological test using antigens from Moscow region, Tula and Ul'janovsk areas. In this study we included wild strains (ultrasonic-treated spirochetes B. garinii H19, B. afzelii P1, B. afzelii P1H13, B. burgdorferi s.s. 39/40, B. burgdorferi s.s. B31), recombinant (expressed in E.coli DbpA, Bgp, Bbk B. garinii, and B. afzelii) antigens and some of their combinations were produced and tested against LD patients and donors serum collected in hospitals of Central regions of Russia by ELISA and Western blotting. Considering sensitivity and specificity, DbpA B. afzelii and DbpA B. garinii recombinant antigens were selected among all probed antigens for regional serology test. As long as DbpA B. afzelii and DbpA B. garinii antigens interacted with LD patient's serum in a complementary mode, it is possible to combine epitopes DbpA B. afzelii and B. garinii in a single antigen for improving sensitivity. We created recombinant fusion protein DbpA B. afzelii/B using dbpA genes from Russian isolates of B. afzelii and B. garinii in E. coli. Fusion DbpA A + G protein was then used for formulation of fast immunochromatographic serodiagnosis test (LF) in a "deep-stick" format. The trials of LF-test were conducted separately at Institute of Rheumatology Russian Academy of Medical Science (using 325 sera) and at the Borreliosis Reference Center of Ministry of Health RF (using 120 reference sera). The average sensitivity and specificity of LF-test was 80.5 and 100 %, respectively.
Background: Staphylococcus aureus is one of the most important human pathogens and causes over 100 nosological forms of diseases. The lack of data on the spread of S. aureus genetic types specific for different forms of staphylococcal infections in Russia makes it difficult to timely identify and control strains of this epidemiologically dangerous bacterial pathogen. Objective: The aim of the study was to carry out a molecular genetic research of S. aureus isolates obtained during a widespread foodborne illness outbreak among builders at the Pulkovo airport in St. Petersburg in 2013. Methods: The ability of the isolates to produce staphylococcal enterotoxins was studied by immunoenzyme techniques. Gene typing was carried out by sequence-specific primer-based PCR, as well as by sequencing genomic nucleotide sequences of two independent isolates of the pathogen. Results: An enterotoxin A gene in genomes of S. aureus isolates etiologically associated with the outbreak was identified. The production of enterotoxin A by the isolates was shown. According to the complex analysis all isolates producing staphylococcal enterotoxins were identical and constituted the S. aureus strain, sequence-type ST30 and spa-type t2509. The genome of the identified S. aureus strain carried a set of various staphylococcal toxins. The full genome sequence among other techniques revealed high levels of similarity between genomes of the strain under study and well-known reference strain S aureus MRSA 252. Conclusion: The complete molecular genetic study of the S. aureus strain involved into the widespread foodborne illness outbreak was first carried out in Russia, allowing of further using the strain as a Russian reference strain to study potential epidemic outbreaks in the Russian Federation.
Despite the fact that the incidence of leprosy in Russia is sporadic, the number of newly identified patients has increased in recent years. In 2017–2018, 4 new cases of leprosy were registered in Russia. The standard methods of research for diagnosis, in addition to the clinical picture, are bacterioscopic study of the skin scarificates and histological study of the skin biopsy specimen. Currently, additional methods are being developed and used to confirm the diagnosis of leprosy, namely, modern serological and genetic diagnostic methods. To use methods such as enzyme-linked immunosorbent assay (ELISA) and membrane immunochromatographic analysis (leprosy LF serotest), it is appropriate to use domestic synthetic mycobacterial antigens (SMA). Key words: Mycobacterium leprae, leprosy, synthetic mycobacterial antigens, PGL-1(phenolic glycolipid-1), LAM (lipoarabinomannan), serodiagnostics, enzyme-linked immunosorbent assay (ELISA), lateral flow (LF) test, BSA (bovine serum albumin)
The glycoconjugates with BSA (bovine serum albumin) were synthesized using a next saccharide: disaccharide derivative M.leprae PGL-1 (phenolic glycolipid-1); a complex of the disaccharide fragment and the branched hexasaccharide fragment LAM (lipoarabinomannan); diarabinofuranose fragment LAM. These glycoconjugates were used as antigenic components for leprosy rapid serotest construction in immunochromatographic format (leprosy LF serotest). The data obtained with sera of leprosy patients, patients who have been in contact with leprosy, and healthy donors indicate that the most promising antigenic component is a BSA conjugate with two synthetic epitopes - a disaccharide derivative of PGL-1 and a branched hexasaccharide fragment of LAM. The leprosy LF serotest with such glycoconjugate demonstrated the greatest diagnostic sensitivity for main forms of leprosy - paucibacillary (PB) and multibacillary (MB)
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