ABSTRACT
A comparative analysis of the detection of CagA-positive strains of H. pylori by immunochromatographic and molecular genetic methods was carried out. We used H. pylori strains isolated from individuals with diseases of the gastrointestinal tract. The immunochromatographic method was implemented using a developed experimental model of an immunochromatographic test system for detecting the H. pylori CagA protein in various biological materials. Determination of the pathogenicity gene cagA of H. pylori was carried out using the «Helikopol SA¼ test system («Litekh¼, Russia). The assessment of the comparability of the results of detecting CagA-positive strains of H. pylori was carried out using statistical methods: Monte-Carlo, calculation of the chi-square test (χ2) and Kendall's τ-b and Somer's d coefficients. Statistical analysis was performed using the software packages «Microsoft Office Excel¼, «Statistica 10.0¼, «WinBUGS 1.4.0.¼ The study showed the absence of a statistically significant difference and the presence of a direct strong correlation between the results of detecting CagA-positive strains by molecular genetic and immunochromatographic methods, which indicates that these methods provide similar results in identifying highly pathogenic strains of H. pylori.
Subject(s)
Helicobacter Infections , Helicobacter pylori , Antigens, Bacterial/genetics , Bacterial Proteins/genetics , Helicobacter Infections/diagnosis , Helicobacter pylori/genetics , Humans , VirulenceABSTRACT
Conducted high-resolution HLA-typing loci HLA-A, -B, -C, -DRB1 and -DQB1 by massively parallel sequencing of 150 potential donors of hematopoietic stem cells from the Republic of Kalmykia. In the studied population, four new alleles identified that not previously registered by the International Committee on the Nomenclature of Factors of the HLA-system of WHO. During the HLA-typing identified: 29 alleles at the HLA-A locus, 44 - at the HLA-B locus, 26 - at the HLA-C locus, 15 - at the DQB1 locus, 37 - at the HLA-DRB1 locus. The following alleles have a frequency of more than 10%: HLA-A*02:01 (11,7%), HLA-A*01:01 (11%), HLA-B*51:01 (10,3%), HLA-B*58:01 (10,3%), HLA-C*06:02 (17,7%), HLA-C*03:04 (10,3%), HLA-C*03:02 (10%), HLA-DQB1*03:01 (26,7%), HLA-DQB1*02:02 (10%), HLA-DRB1*07:01 (11,7%). The most common HLA-A-B-C-DQB1-DRB1 haplotype is A*02:05-B*50:01-C*06:02-DQB1*02:02-DRB1*07:01 (3,7%). Deviations from the Hardy - Weinberg equilibrium not identified.
Subject(s)
Alleles , Ethnicity/genetics , HLA-A Antigens/genetics , HLA-B Antigens/genetics , Haplotypes , Gene Frequency , HLA-C Antigens , Hematopoietic Stem Cells , Humans , RussiaABSTRACT
Using data obtained from domestic and foreign sources, we formed a set of primers and fluorogenic probes for analyzing twentysix specific sequence polymorphisms and one reference gene. In the course of evaluating the effectiveness of real-time PCR, using the example of one of the markers (S01a), we obtained the optimal amount of DNA per reaction (70 ng), providing a resolution of at least 0.1% of the method with the ability to estimate linear chimerism. Formed panel of primers for genetic polymorphisms - InDel has a high degree of informational content for donor-recipient pairs of Russia. From January 2018 to June 2019, a quantitative assessment of the level of linear (CD3 +, CD34 +) and general chimerism was carried out for 28 patients of the clinic of the Institution. Finally, we analyzed patients who received allografts and present 4 different clinical situations that illustrate the informativity level of this method.
Subject(s)
Chimerism , Real-Time Polymerase Chain Reaction , Stem Cell Transplantation , Humans , INDEL Mutation , Polymorphism, Genetic , RussiaABSTRACT
The immunochromatographic test-system was developed for detecting protein of pathogenicity of CagA Helicobacter pylori in various biological samples (feces, content of dento-gingival recesses) and also in culture of H.pylori. The test-system represents multi-membrane composite on the basis of membranes manufactured by "MDI" (India). The main immunochemical components of test-system are conjugate of nanoparticles of colloid gold with size of 30 nm with monoclonal antibodies (clone HP-387), applied to membrane for conjugate; monoclonal antibodies (clone HP-1811) and anti-species antibodies of goat against Ig of mouse, applied to nitrocellulose membrane correspondingly in test and control zones. All antibodies are produced by the firm "Bialeksa" (Russia).
Subject(s)
Antigens, Bacterial/isolation & purification , Bacterial Proteins/isolation & purification , Helicobacter Infections/immunology , Helicobacter pylori/pathogenicity , Immunoassay , Animals , Antibodies, Bacterial/immunology , Antibodies, Monoclonal/immunology , Mice , VirulenceABSTRACT
The article summarizes data of research studies concerning searching optimal conditions of development of highly specific and highly sensitive immune chromatography test-systems designed for detection of agents of infectious diseases. The analysis was implemented concerning choosing of optimal size of nano-particles of colloid gold and concentration of antibodies for production of conjugate. The alternatives are presented concerning the most frequently used combinations of buffer solutions applied for development and analysis of immune chromatography test-systems. The preferences are established in the field of characteristics of membranes used in composition of multi-membrane composite of immune chromatographic test-systems.
Subject(s)
Chromatography , Communicable Diseases/immunology , Immunoconjugates/immunology , Nanoparticles/chemistry , Antibodies/immunology , Communicable Diseases/diagnosis , Gold Colloid/chemistry , Humans , Immunoconjugates/isolation & purification , Membranes, ArtificialABSTRACT
The article describes the experience with the treatment of 886 patients with lactation mastitis. For limited purulent foci preference is given to the dissection or wide opening of the abscess with wound dialysis and putting primary sutures. In patients with mastitis and symptoms of anaerobic damage wide openings (beyond the limits of the tissue involved) with necrectomy and following aeration of the wound are recommended; after complete cleansing the wound secondary sutures are possible or plastic closure of the wound. Complex conservative treatment after operation must be combined with the application of trichopol and its derivatives, oxygentherapy.