Clinical-epidemiological features and improvement of botulism laboratory diagnos-102 tics in Irkutsk region

Irkutsk Region is one of the Russian Regions where botulism cases are registered yearly. Botulism morbidity in Irkutsk Region is analysed for the ten-year period (since 2002); its clinical-epidemiological features are revealed at the present stage. Sporadic cases of botulism, are dominated. (81,9 %). From. 2006 in Irkutsk Region botulism share makes 100 % in food poisoning number. A considerable role in the infection transmission plays consumption of fish, mainly smoked or salty Baikal omul (90,9 %). In 30 % of cases injured people purchased fish at private persons in unascertained trade places, 60 % of patients ate own salting fish. In Irkutsk Region botulism is registered more often from July till November with peaks in July (58,3 %) and. September (20,8 %) that is connected, with activation of unprofitable fabricated fish selling in combination, with adverse conditions of this foodstuff storage. Botulism affects various age groups of population with, prevalence of the age category 30-49 years old. (47,8 %). Socially active able-bodied population. (72,3 %) is most often affected that increases the social importance of the disease. Mainly, moderate severe (56, 5 %) and. severe (34,8 %) disease forms were registered. Primary symptoms include two variants - dyspeptically-paralytic and. ophthalmologic-neurological forms. Analysis of botulism etiological structure showed, that the leading part in the disease occurrence in Irkutsk Region belonged to botulinum. type Е (73, 9 %). Clinical material and. foodstuff testing to botulinum toxin presence was conducted in parallel by the standard reaction of biological toxin neutralisation in white mice and dot-immunoanalysis with application of the test system that we have developed. Possibility of dot-immunoassay application in laboratory diagnostics of botulism is estimated as the express method with high specificity, sensitivity and. simplicity

Improvement of a test-system for the botulinus toxin screening in dot-immunoanalysis

We constructed a test system for dot-immunoassay (DIA) to accelerate definition and identification of botulinus toxins and also to refuse from application of laboratory animals for routine screening of clinical samples, foodstuff and environments. This system permitted to detect botulinus toxin during approximately 2 h in the tested samples. Sensitivity of this DIA in some cases exceeded the mice biotest. This improved method has minimum reaction to nonspecific exposures from the investigated biological substrata. It is simple to conduct. It is high efficient and expressive, does not require to use expensive equipment and the reactants, special training for the personnel. Lyophilization conditions for the immune reagents used for the test system preparation for botulinus toxin dot-immunoassay were selected. High sensitivity, specificity of the analysis are remained, stability of the preparations (periods of storage) is increased. This method is convenient to use in field conditions at extreme situations, in particular, in mobile autolaboratories for epidemiological survey

The production of hyperimmune pseudotuberculosis sera

Background. Pseudotuberculosis remains a serious public health problem, which dictates the need to develop simple and rapid diagnostic methods for this disease. The effectiveness of the latter largely depends on the activity and specificity of the diagnostic sera. Currently, in our country, a diagnostic serum for Yersinia pseudotuberculosis O-monovalent (serotype I, III) is produced with a regulated area of application: for an approximate agglutination reaction on glass (Pasteur NIIEM, St. Petersburg). Preparation of pseudotuberculosis sera for a wider scope of their use, in particular, as a  source of specific antibodies in the design of diagnostic immunobiological preparations and test systems for pseudotuberculosis, is relevant and in demand in healthcare practice.Aims. To obtain hyperimmune pseudotuberculosis sera, promising for use in the practice of laboratory studies for pseudotuberculosis.Materials and methods. Chinchilla rabbits aged 3–6 months, weighing 2.5–3.0 kg served as animal producers of pseudotuberculosis sera. As immunogens, we used a corpuscular antigen (CAg) - a suspension of cells of the epidemically significant Yersinia pseudotuberculosis 3704 O: 1b strain isolated by an outbreak of pseudotuberculosis in the town of Zima, Irkutsk region, inactivated by boiling, and a preparation obtained from the outer membranes, containing the main surface immunogen of the strain. For the adsorption of experimental sera, in order to minimize the nonspecific response, we used heterologous microorganisms Escherichiacoli, Salmonella typhimurium, Shigella flexneri and Y. enterocolitica O:3, which have a similarity of surface antigenic structures with the causative agent of  pseudotuberculosis. The presence of specific antibodies in experimental pseudotuberculosis sera was determined in a volumetric agglutinationreaction.Results. Optimal schemes of rabbit immunization were selected, which made it possible to obtain hyperimmune sera against Y. pseudotuberculosis 3704 O:1b antigens with an agglutination activity of 1:3200-1:6400. In low dilutions of experimental sera (1:100–1:400), obtained against corpuscular antigen and outer membrane antigens (Escherichia coli, Salmonella typhimurium, Shigella flexneri , Y. enterocolitica О:3, S . enteritidis Gartnery) was observed in the agglutination reaction (AR). After the adsorption of experimental pseudotuberculosis sera by cells of heterologous strains, there was no cross-reaction with the indicated microorganisms in the  agglutination reaction.Conclusions. The obtained hyperimmune adsorbed sera against the boiling-inactivated Y. pseudotuberculosi s 3704 O: 1b corpuscular antigen and outer membrane antigens can be used as a source of specific antibodies in the design of diagnostic immunobiological preparations for the detection of pseudotuberculosis microbe, as well as in the monitoring of the epidemic situation

Using dot-immunoassay in decoding the outbreak of pseudotuberculosis in the Tomsk region

Background. Pseudotuberculosis remains a serious healthcare problem, which determines the expediency of developing the express methods for its early diagnosis. To detect the pathogen, we designed test system for dot-immunoassay (DIA) based on antibodies labeled with silver nanoparticles (SNPs) isolated from hyperimmune rabbit serum obtained against killed cells of  Yersinia pseudotuberculosis of O:1b serovariant.The aim. To assess the possibility of using dot-immunoassay for express identification of Y. pseudotuberculosis cultures isolated from clinical material and environmental objects at the initial stage of bacteriological study during laboratory diagnosis of the disease.Methods. We used the materials from the outbreak of pseudotuberculosis in the Krylovskaya Boarding School of the Bakcharsky district of the Tomsk region in 2021. Specific antibodies from hyperimmune rabbit sera obtained against Y. pseudotuberculosis 3704 particulate antigen of O:1b serotype were labeled with SNPs and used in DIA on nitrocellulose membranes with visualization of reaction results with a solution of a physical developer. The presence of the causative agent of pseudotuberculosis in the test material was inferred by the formation of gray spots of different intensity (from 4+ to 1+).Results. All Y.  pseudotuberculosis strains isolated using bacteriological method on  the second day of the study from clinical material obtained from sick people and environmental objects were detected in DIA at concentrations ≥ 3.1 × 104 microbial cells per milliliter (m.c./ml).Conclusion. The designed test system for dot-immunoassay using SNPs as a marker of specific antibodies for the detection of Y.  pseudotuberculosis in cultures isolated from swabs from vegetables and clinical material from patients, including those  with  mixed infection, allows us to  detect a specific corpuscular antigen with a high sensitivity (≥ 3.1 × 104 m.c./ml), providing express identification of isolated cultures at the initial stage of bacteriological study

Новые данные о семействе Blaberidae (Dictyoptera) из Юго-Восточной Азии: новые виды, морфологическое разнообразие и филогения на основании последовательностей рибосомальной ДНК

New cockroaches of the family Blaberidae are described from Southern Sumatra: two new spe- cies of the genus Cyrtonotula Uvarov, 1939, C. secunda sp. nov. and C. tertia sp. nov. (Epilam- prinae); and one new species of the genus Paranauphoeta J.W.H. Rehn, 1951, P. pullata sp. nov. (Paranauphoetinae). Detailed morphological descriptions of the new species are given. Structures of the male genitalia of the genus Cyrtonotula are described for the first time. Hypothesis on the relationships of these new taxa as well as Morphna maculata Brunner von Wattenwyl, 1865, Rhabdoblatta sp. and Pseudophoraspis sp. based on 28S ribosomal DNA sequences is discussed

Новые данные о семействе Blaberidae (Dictyoptera) из Юго-Восточной Азии: новые виды, морфологическое разнообразие и филогения на основании последовательностей рибосомальной ДНК

New cockroaches of the family Blaberidae are described from Southern Sumatra: two new spe- cies of the genus Cyrtonotula Uvarov, 1939, C. secunda sp. nov. and C. tertia sp. nov. (Epilam- prinae); and one new species of the genus Paranauphoeta J.W.H. Rehn, 1951, P. pullata sp. nov. (Paranauphoetinae). Detailed morphological descriptions of the new species are given. Structures of the male genitalia of the genus Cyrtonotula are described for the first time. Hypothesis on the relationships of these new taxa as well as Morphna maculata Brunner von Wattenwyl, 1865, Rhabdoblatta sp. and Pseudophoraspis sp. based on 28S ribosomal DNA sequences is discussed

The multicenter experience with bendamustine in the treatment of relapsed and refractory multiple myeloma

Relapsed and refractory (R/R) multiple myeloma (MM) constitutes a specific and unmet medical need. Median survival ranges from as little as 6 to 9 months, and responses to treatment are characteristically short. In patients with R/R MM after therapy of bortezomib and/or immunomodulators a bendamustine-based treatment can be used as “salvage”.In this retrospective analysis we have identified 32 patients with R/R MM by means of case research, have been bendamustine-based treated at Hematological Clinics of Russian Federation since 2011. Median age was 67 (43–81) years, the female/male ratio was 2.5:1. After in median 2 (1–7) lines of prior therapy patients received in median 3 (1–9) cycles of bendamustine-based therapy. Bendamustine dosage was 70–120 mg/m2 /day on 2 days of each 28-day cycle until progressive disease or intolerability. Overall rate response was 56.2 %: 21.9 % partial response, stable disease 34.4 %. Median time to progression was 5.3 (0.8–18.0) months and median overall survival was 25.4 (0.8–47.1) months. Hematologic toxicity was in 53.2 % of patients

COMPARATIVE ANALYSIS OF EFFECTIVENESS OF SOLID-PHASE METHODS OF IMMUNE DETECTION OF BOTULINIC TOXIN IN BLOOD SERA OF A PATIENT WITH BOTULISM DIAGNOSIS

Aim. Comparison of effectiveness of solid phase methods of immune detection of botulinic toxin in blood sera of a patient with botulism diagnosis: dot-immune assay using specific anti-botulinic antibodies (AT) labeled with nanoparticles of colloid silver, phosphorescent analysis (PHOSPHAN) using streptavidin label with platinum coproporphyrin (PtCP) and polystyrene nanoparticles, containing chelate complex of europium ions with naphthoyl trifluoroacetone (NA-Eu). Materials and methods. Silver nanoparticle labeled IgG isolated from a commercial diagnostic polyvalent sera against type А, В, С, E, F botulotoxins manufactured by SPA Allergen (Stavropol) with 5000 - 10000 IU activity and biotin conjugated commercial monoclonal antibodies against botulotoxin A, polyclonal mono-specific AB against botulotixin В and E and polyvalent immunoglobulin against botulotoxin А, В, С, E, F. Detection ofbotulotoxin in clinical material was carried out in dot-immunoassay on nitrocellulose membrane by PHOSPHAN method in an experimental test system using 2 detector systems based on streptavidin: PtCP and NA-Eu. Results. Botulotoxin was detected in blood sera of the botulism patient using both of the developed immune detection methods. PHOSPHAN method allowed to identify serotype В botulotoxin, that corresponded with the results obtained in botulotoxin biological neutralization reaction. Sensitivity of PHOSPHAN with NA-Eu luminescent nanoparticle based detection system was higher than with PtCP label. Conclusion. The developed methods (PHOSPHAN and dot-immunoassay) differ by high specificity and sensitivity and may be recommended for express detection of botulinic toxin in clinical material