Diagnostic Sera Research Articles

Clinical Characteristics, Serotypes and Antimicrobial Resistance of Invasive Salmonella Infections in HIV-Infected Patients in Hangzhou, China, 2012-2023.

Developing countries, invasive Salmonella infections can cause considerable morbidity and mortality. There is a relative lack of data on coinfection with Salmonella in HIV-infected patients in Hangzhou, China. In this study, we manually collected case data of patients aged >18 years with HIV combined with invasive Salmonella infections admitted to Xixi Hospital in Hangzhou from January 2012 to August 2023 by logging into the Hospital Information System, and identified 26 strains of invasive Salmonella using a fully automated microbiological identification system and mass spectrometer. Serotypes were determined using Salmonella diagnostic sera based on the White-Kauffmann-Le Minor scheme. Drug sensitivity tests were performed using the automated instrumental method of the MIC method. A total of 26 HIV-infected patients with invasive Salmonella coinfections were identified over 11 years; Twenty-five of the 26 patients (96.2%) were males, with a mean age of 33.5 years (26.75, 46.75). The most common type of infection was bloodstream infection (92.3%). One patient also had concomitant meningitis and osteoarthritis, followed by pneumonia (7.7%). The presence of multiple bacterial infections or even multiple opportunistic pathogens was clearly established in 7 (26.9%) patients. Three (11.6%) patients were automatically discharged from the hospital with deterioration of their condition, and one (3.8%) patient died. Salmonella enteritidis was the most common serotype in 6 patients (23.2%), and Salmonella Dublin was the most common serotype in 6 patients (23.2%). Drug sensitivity results revealed multidrug resistance in a total of 8 (30.8%) patients. The clinical presentation of invasive Salmonella infection in HIV patients is nonspecific and easily masked by other mixed infections. A CD4+ count <100 cells/µL and comorbid intestinal lesions may be important susceptibility factors. Salmonella has a high rate of resistance to common antibiotics, and the risk of multidrug resistance should not be ignored.

Obtaining and Characterization of Experimental Diagnostic Brucellosis Rabbit Sera for the Development of a National Standard for Brucellosis Serum

Relevance. The production of brucellosis diagnosticums is currently being established in the medical and veterinary practice of the Republic of Uzbekistan. However, due to the lack of a national standard serum in the country and the high cost of analog standard serums on the market, problems may arise in the production of diagnostic tools. In addition, the lack of national standard serums causes problems in quality control of commercial and local antigens, hinders the development of local drugs – brucellosis diagnostics, which reduces the effectiveness of serological diagnosis of brucellosis in humans and animals. Given the above, it becomes necessary to develop and produce a standard serum with a high level of safety, which will improve the methods of laboratory diagnosis of various forms of brucellosis in patients and animals to identify hidden foci of infection in Uzbekistan. The aim of the study is to obtain and characterize experimental standard diagnostic brucellosis rabbit sera for the development of national and industry standard serum with appropriate titers. Materials and Methods. The experiments were carried out in full compliance with the European Council Directive 2010/63/EU on compliance with ethical principles in working with laboratory animals and the experiments were carried out on the basis of the methodological manual «Methods and rules of work» approved by the Ministry of Health of the Republic of Uzbekistan in 2016 with laboratory animals in experimental microbiological and immunological studies. To obtain positive standard sera against brucellosis pathogens, 12 rabbits weighing from 2.2 kg to 4.6 kg, aged 6 to 12 months, were used. The animals were kept under identical conditions on a standard vivarium diet. For the experiment, animals adapted to the conditions of the study were selected and kept in quarantine for 21 days. The maintenance and conduct of the studies followed international rules for the humane treatment of animals. Results and Discussion. During the evaluation of the heterologic specificity of the obtained sera for 23 collection and hospital strains, a weakly positive response (+) was observed in 16.7% of cases. Rabbit sera positive for heterologic specificity were adsorbed on suspensions of inactivated microorganisms (Citrobacter spp. and Enterobacteriaceae spp.) - no significant drop in titer was observed, and cross-agglutination was eliminated. A significant increase in the total protein level was noted after the first and fourth immunizations. Before the first immunization, the total protein level was 59.57 g/l, and after the fourth immunization, it increased threefold to 157.17 g/l. There was also an increase in albumin and globulin (33.2 g/L and 26.37 g/L, respectively) to 79.37 g/L and 77.43 g/L, respectively. IgM levels remained almost unchanged, increasing 3.5-fold after the second immunization and decreasing to 0.24 mg/ml after the fourth immunization. An increase in the level of IgG after the second and third immunizations was revealed, followed by a decrease after the fourth immunization. Conclusion. The obtained national and industry standard diagnostic polyvalent brucellosis sera, as well as the developed methods of laboratory control of its quality and efficiency testing will make it possible to produce candidate standard serum samples for agglutination reaction test and detection of Brucella strains. The obtained standard serum samples will be further used for control of the produced immunobiological diagnostic preparations and provision of unified requirements to their quality. The goal was achieved – improving the serological diagnosis of brucellosis, laying the foundation for further study and standardization of serum.

DIPG-19. SURGICAL STATUS IN THE ERA OF NEW MOLECULAR DIAGNOSTICS: INSIGHTS FROM A SINGLE-CENTER CASE SERIES REPORT IN CHINA

Abstract The therapeutic value of surgery for DIPG has been a subject of controversy. Surgical biopsy provides valuable information on molecular markers of brain gliomas, aiding in further personalized treatment. This retrospective study aimed to elucidate the real-world application of surgery in Diffuse Intrinsic Pontine Glioma (DIPG) in the era of new molecular diagnostics. This study from Guangdong Sanjiu Brain Hospital investigates the surgical landscape in the treatment of Diffuse Intrinsic Pontine Glioma (DIPG) amid the new molecular diagnostics era (Jan 2017 to Feb 2023). Among 184 cases, 58.6% underwent surgery. Notably, 97.2% were introduced to surgery, with 65.8% opting for open surgeries and 34.2% utilizing the Rosa robot. Patients aged 18 and above showed a higher surgery rate (78.3%) than those below 18 (49.1%), revealing a significant difference (P&amp;lt;0.05). The median time from surgery to radiotherapy varied between open (19 days) and stereotactic biopsy (7 days). Complications included increased cranial nerve dysfunction in 20 open biopsy cases. All 108 patients received pathological diagnoses, with 79 having diffuse midline gliomas. Targeted therapy based on second-generation sequencing was administered to seven patients, involving drugs like ribociclib and rigosertib. The study concludes that compared to a decade ago, there is an increased willingness among oncologists to perform brainstem surgery, especially in older patients. Stereotactic biopsy, with fewer side effects, allows swift initiation of radiotherapy. Obtaining lesion tissue during surgery holds promise for new treatments and future research.

Application of alternative identification methods for live tularaemia and brucellosis vaccines

SCIENTIFIC RELEVANCE. The sanctions currently imposed on the Russian Federation requireusing Russian diagnostic products in vaccine quality assessments, as well as searching for alternative testing methods, in particular, serological ones.AIM. This study aimed to demonstrate the possibility of using alternative serological methods, including immunoenzymatic, direct agglutination, and indirect haemagglutination methods, for the identification of tularaemia and brucellosis vaccines in studies and quality assessments. MATERIALS AND METHODS. This study used the established pharmacopoeial reference standard (RS) for the bacterial suspension opacity of 10 international opacity units, the established pharmacopoeial RS for and two commercial batches of the live brucellosis vaccine (Brucella abortus 19 BA), and the established and candidate pharmacopoeial RSs for the live tularaemia vaccine (Francisella tularensis 15 NIIEG). These pharmacopoeial RSs were certified by the Scientific Centre for Expert Evaluation of Medicinal Products. Serological testing used Russian commercial diagnostic products, including dry diagnostic sera (polyvalent brucellosis and tularaemia sera for agglutination tests), a liquid erythrocytic diagnostic preparation of tularaemia immunoglobulin, and enzyme immunoassay (ELISA) diagnostic kits for detecting the causative agents for brucellosis and tularaemia. Statistical analysis involved using Microsoft Excel (P=0.95) for ELISA results and qualitative evaluation for the results of direct agglutination and indirect haemagglutination tests.RESULTS. All the tested batches demonstrated positive results. Live brucellosis vaccine batches showed positive results in the slide agglutination tests, while live tularaemia vaccine batches yielded positive results in the tube agglutination tests. All indirect haemagglutination tests showed haemagglutination in live tularaemia vaccine samples at the same concentration as positive control samples (6.25×106 cells/mL). ELISA tests showed similar optical density values (D)for the two vaccines and positive control samples. Live tularaemia and brucellosis vaccines (undiluted, 1.0×109 cells/mL) had D=2.133±0.273 and D=0.127±0.013, whereas the corresponding control samples had D=1.942±0.056 and D=0.123±0.007, respectively. The results reflected the presence of brucellosis or tularaemia microbes in the samples, confirming the identity of the vaccines.CONCLUSIONS. Serological immunoenzymatic, direct agglutination, and indirect haemagglu tination methods with Russian diagnostic products can be used to identify live brucellosis and tularaemia vaccines during quality assessment. The agglutination method with Russian diagnostic sera can be recommended as an alternative quality assessment method for the identification of live brucellosis and tularaemia vaccines, as this method offers time efficiency, simple visual evaluation of results, and low costs and relatively long shelf lives of diagnostic products. However, ELISA and indirect haemagglutination methods cannot be recommended for this purpose because of their labour-intensive and uneconomical nature. The results of this study may support the introduction of the agglutination method in the regulatory documents for live brucellosis and tularaemia vaccines (as an alternative method).

Использование бесклеточных антигенных фракций возбудителя псевдотуберкулёза для получения диагностических сывороток

The development of simple, express, and at the same time sensitive and selective methods for diagnosing pseudotuberculosis remains an urgent task of practical public health. The effectiveness of the test systems largely depends on the activity and the specificity of a diagnostic pseudotuberculous sera. Achieving high sensitivity and specificity of diagnostic test systems largely depends on the presence of specific pseudotuberculous sera with a high antibody titer, which, in turn, is ensured by the proper selection of the animal producer, the physicochemical properties of the immunogen, its dose, multiplicity and duration. introduction and use of adjuvants. Currently, diagnostic serum for Yersinia pseudotuberculosis for an approximate agglutination test on a glass slides is produced in the Russian Federation (FSUE SPbNIIVS FMBA of RUSSIA). Obtaining of pseudotuberculous sera for a wider scope of their use, in particular, in the design of diagnostic immunobiological preparations and test systems for pseudotuberculosis, is relevant and in demand in healthcare practice. The purpose of this study was to obtain and characterize hyperimmune specific sera against cellfree antigenic fractions of the causative agent of pseudotuberculosis. Chinchilla rabbits were used as sera producers. Cell-free antigenic fractions (urea extract, outer membranes, and proteinlipopolysaccharide complex) obtained by treating Yersinia pseudotuberculosis 3704 O:1b strain cells with a 9 M urea solution followed by differential centrifugation were used as immunogens. The presence of specific antibodies in experimental pseudotuberculosis sera was determined in the test tube agglutination reaction, passive hemagglutination reaction and agarose gel double immunodiffusion test. The proposed schemes of immunization of rabbits with subcellular fractions of the pseudotuberculosis microbe make it possible to prepare hyperimmune sera with activity in the standard tubeagglutination test ≥1:1600, in the indirect hemagglutination test ≥1:3200. There was no crossreactivity of antibodies in the immunodiffusion test with heterologous, closely related antigenically microorganisms (Yersinia enterocolitica 628, Salmonella typhi 21, Shigella flexneri 1964). The most important antigenic material is concentrated in the surface structures of pathogens of bacterial infectious diseases, against which, during immunization of laboratory animals, specific antibodies are produced that can interact with antigenic epitopes of outer membrane proteins and O-specific side chains of lipopolysaccharide (O-antigen) exposed on the surface of the microorganism. The data obtained confirm the effectiveness of the use of surface cell-free antigenic fractions of the pathogen to produce highly active specific pseudotuberculosis sera, suitable for use as a source of specific antibodies in the design of diagnostic immunobiological preparations and test systems for pseudotuberculosis.

Epidemiological Analysis of the Swine Perfringens Disease in the Aragatsotn Region

Over the recent 20 years, for the first time, the disease of swine perfringens has been studied in the Aragatsotn region of RA and related epidemiological analysis has been conducted. To identify A, C, and D serological types of Cl. Perfringens and special diagnostic sera produced by the Kursk biofactory were used. It has been found that the main serological infection type of swine perfringens is C and sometimes A and D subtypes. In the pig breeding farms of the mentioned region about 6887 heads of swine (aging from 3 days to 1 year old) were registered. Per the research results, about 45 out of 72 communities were recognized as insecure/vulnerable. As to the above-stated animal number, it should be mentioned, that 1722 heads of swine fell down, whereas 5510 were considered bacteria carriers. Thus, as a result of epidemiological analysis, the morbidity (bacteria carrying rate), mortality, insecurity/vulnerability, and focality coefficients (indicators) have made 0.80 (80 %), 0.25 (25 %), 0.63 (63 %), and 122 animals, respectively.

A qualitative investigation of paediatric intensive care staff attitudes towards the diagnosis of lower respiratory tract infection in the molecular diagnostics era

BackgroundIn the past decade, molecular diagnostic syndromic arrays incorporating a range of bacterial and viral pathogens have been described. It is unclear how paediatric intensive care unit (PICU) staff diagnose lower respiratory tract infection (LRTI) and integrate diagnostic array results into antimicrobial decision-making.MethodsAn online survey with eleven questions was distributed throughout paediatric intensive care societies in the UK, continental Europe and Australasia with a total of 755 members. Participants were asked to rate the clinical factors and investigations they used when prescribing for LRTI. Semi-structured interviews were undertaken with staff who participated in a single-centre observational study of a 52-pathogen diagnostic array.ResultsSeventy-two survey responses were received; most responses were from senior doctors. Whilst diagnostic arrays were used less frequently than routine investigations (i.e. microbiological culture), they were of comparable perceived utility when making antimicrobial decisions. Prescribers reported that for arrays to be clinically impactful, they would need to deliver results within 6 h for stable patients and within 1 h for unstable patients to inform their immediate decision to prescribe antimicrobials. From 16 staff interviews, we identified that arrays were helpful for the diagnosis and screening of bacterial LRTI. Staff reported it could be challenging to interpret results in some cases due to the high sensitivity of the test. Therefore, results were considered within the context of the patient and discussed within the multidisciplinary team.ConclusionsDiagnostic arrays were considered of comparable value to microbiological investigations by PICU prescribers. Our findings support the need for further clinical and economic evaluation of diagnostic arrays in a randomised control trial.Trial registrationClinicaltrials.gov, NCT04233268. Registered on 18 January 2020.

Assessment of the Role of Free-Living and Farmed Fallow Deer (Dama dama) as A Potential Source of Human Infection with Multiple-Drug-Resistant Strains of Yersinia enterocolitica and Yersinia pseudotuberculosis.

Yersinia enterocolitica and Y. pseudotuberculosis are Gram-negative, facultative anaerobic bacteria that cause yersiniosis-one of the most important zoonotic diseases of the digestive tract. The aim of this study was to determine the prevalence of potentially human-pathogenic Y. enterocolitica and Y. pseudotuberculosis strains in free-living and farmed fallow deer, and to evaluate their sensitivity to chemotherapeutics. A total of 372 rectal swabs were analyzed, including 262 from free-living and 110 from farmed fallow deer. Due to the psychrophilic properties of Yersinia, two samples were collected from each animal. Seven Y. enterocolitica strains were isolated from free-living fallow deer, while two strains were isolated from farmed fallow deer. Yersinia pseudotuberculosis strains were not identified. All isolated Y. enterocolitica strains were ystB-positive, and phylogenetic analysis based on the nucleotide sequences of this gene revealed the presence of two phylogenetic groups. Yersinia enterocolitica strains isolated from fallow deer belonged to biotype 1A, and serotyping analysis demonstrated that the vast majority did not agglutinate with any diagnostic sera. All strains were multiple drug resistant and were not sensitive to at least four of the tested chemotherapeutics (amoxicillin with clavulanic acid, ampicillin, cefalexin, and streptomycin). One Y. enterocolitica strain isolated from a free-living animal was resistant to nine out of the 13 analyzed chemotherapeutics and was intermediately sensitive to the four remaining chemotherapeutics. The highest sensitivity was noted in case of ciprofloxacin (five strains) and trimethoprim-sulfamethoxazole (three strains). Only one strain isolated from a free-living animal was sensitive to three out of the 13 examined antibiotics, whereas the remaining strains were sensitive to only one drug or were not sensitive to any of the chemotherapeutics used. The results of this study indicate that multiple drug-resistant Y. enterocolitica strains can be carried by free-living and farmed fallow deer. This observation gives serious cause for concern because the meat of fallow deer and other ruminants is often consumed semi-raw (steak) or raw (tartar steak).

Study of the Surface Antigenic Determinants of &lt;i&gt;Vibrio cholerae&lt;/i&gt; Strains with Atypical Agglutinability Using the Panel of Monoclonal Antibodies

The aim of the work was to study surface antigenic determinants of V. cholerae R-variant strains using enzyme immunoassay and a panel of monoclonal antibodies (MAbs).Materials and methods. 60 strains of V. cholerae R-variant isolated from ambient environment objects in the territories of the former USSR and the constituent entities of the Russian Federation over a 30-year period (1988–2019) were investigated in the slide agglutination reaction with cholera diagnostic sera, enzyme immunoassay (ELISA) using the panel of MAbs specific to membrane proteins and a set of reagents “Monoclonal diagnostic immunoglobulins labeled with horseradish peroxidase, dry, for serological identification of V. cholerae O1 and O139 (in vitro) through ELISA and dot-ELISA”.Results and discussion. The analysis of the surface structures of V. cholerae R-variant strains with atypical agglutinability has been carried out applying enzyme immunoassay. It showed that individual strains with different amounts of O-antigen are registered among the studied strains identified at isolation as V. cholerae R-variant (the optical density range is from 0.261±0.002 to 1.312±0.003). Epitopes of specific O-antigen were found in some “conservative” strains (30 %) that are agglutinated only with RO serum, and in several strains (20 %) that do not have the wbeT gene that determines its synthesis, and lost agglutinability with all diagnostic cholera sera, including RO. The protein epitopes recognized by complementary MAbs are represented with varying frequency in the composition of surface antigens of R-vibrios; a decrease in their representation or absence on the cell surface correlates with the modification or loss of R-LPS and is accompanied by a negative agglutination reaction.

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 agglutination reaction. 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.

Токсигенність штамів yersinia еnterocolitica, ізольованих з різних об’єктів

Токсигенність штамів yersinia еnterocolitica, ізольованих з різних об’єктів

The Activity of Influenza Viruses during 2017-2018 Season in Russia and Countries of the Northern Hemisphere: Conflict by the B-virus Vaccine Component

Relevance. Influenza viruses, having an extremely high variability of the genome and significant environmental plasticity, continue to maintain a potential threat to the biological safety of mankind. The purpose Aims of the work is to analyze the features of the epidemic season of2017-2018. Materials and methods . The collection of data on the incidence and laboratory diagnosis of influenza and ARVI was carried out in the framework of the epidemiological surveillance of the circulation of influenza viruses in the Russian Federation. The observation period was from 40 weeks (october) 2017 to 25 weeks (june) 2018. The typing of the isolates was performed in the reaction of inhibition of hemagglutination activity (HI) according to the standard technique with diagnostic sera to reference and epidemic influenza viruses. Results and discussion . The article presents the features of the influenza virus circulation n for the period from october 2017 to may 2018 in some territories of Russia, collaborating with D. I. Ivanovsky Institute of Virology. The epidemic season had its own peculiarities: delayed and long-term activity of three influenza viruses, the share participation of which was almost equal, with some dominance of the influenza A(H1N1)pdm09 virus by the end of the season. Morbidity rates for the total population were comparable with the last epidemic season, meantime the morbidity among schoolchildren was higher. The number of hospitalizations and lethal outcomes was lower and mostly was found in patients of 65 years and older. In patients with severe acute respiratory infection (SARI) influenza A(H1N1)pdm09 was detected more frequently (58%). The influenza A(H1N1)pdm09 and A(H3N2) viruses were antigenically similar to strains included in influenza vaccines, at the same time, 96% of the isolated strains of influenza B virus belonged to a different evolutionary line. Epidemic strains were sensitive to neuraminidase inhibitors, except for 5 strains of influenza A(H1N1)pdm09 virus, isolated from pregnant women. Conclusions. The activity of non- influenza ARI viruses was similar to preliminary epidemic seasons. Recommendations on the influenza vaccines composition for the Northern hemisphere for 2019-2020 season are presented as well.

DEVELOPMENT OF SCIENTIFICALLY BASED APPROACHES TO MANAGEMENT OF THE PROCESSES OF CONSERVATION OF THE ACTIVITY OF DIAGNOSTIC SERA OF CASES WITH IN LARVAL PARASITIC DISEASES

There was performed a study of the diagnostic activity of sera with antibodies to Toxocara spp. and Echinococcus spp. in conditions of long storage at different temperature regimes. The use of cryoprotectants makes it possible to increase the safety of diagnostic sera by two times during prolonged storage at low temperatures.

COMPARATIVE CHARACTERISTICS OF THE EXPERIMENTAL AGGLUTININS ACTIVITY OF IMMUNE SERUM AGAINST STRAINS OF BURKHOLDERIA PSEUDOMALLEI AND BURKHOLDERIA MALLEI ISOLATED IN DIFFERENT REGIONS

Burkholderia pseudomallei and Burkholderia mallei are the causative agents of melioidosis and glanders, dangerous infectious diseases. These microorganisms are not endemic for the central zone of the Russian Federation, but the possibility of an imported case or deliberate use as a biological weapon is not ruled out. In practical work for accelerated detection and subsequent identification of pathogenic microorganisms, the method of orienting reaction of slide-agglutination is most often used as the simplest, requiring no additional equipment and easy enough to perform. Since in Russia standard (production) diagnostic species-specific sera are not produced for any Burkholderia species, experimental series of preparations are used in the conditions of specialized laboratories for setting diagnostic immunological reactions. In the present work we investigated four types of experimental rabbit immune serum using strains of pathogens melioidosis and glanders collection of Volgograd Research Plague Control Institute. The most active was serum obtained against live B. pseudomallei VPA cells, the least is serum against B. pseudomallei C-141; with complex serum against cells of 5 strains of Burkholderia thailandensis and with serum against extracellular antigen B. thailandensis 264 agglutination was observed only at low dilutions.

ENTEROVIRALES AS A REASON OF INTRAUTERINE INFECTIONS OF FETUS AND NEONATAL DISEASES

Introduction. Intrauterine infections remain an urgent issue of modern Perinatology and belong to the group of diseases diagnosis of which is associated with some difficulties. The number of causative agents of intrauterine infections in case of ante-, intra- and perinatal infectionis practically unlimited. However, one of the first places belongs to pathogens of viral infections with enteroviruses being of the most interest, as a transplacental (vertical) way of infection the fetus and neonates was proved. The significance of enteroviruses in the pathology of the fetus and neonate is unknown, since there is no verification ofserotypes of enteroviruses in pregnant women and neonates.The objective is to determine the effect of enteroviruses on the fetus and neonate based on the verification of circulating enteroviruses serotypes, the definition of risk factors of the perinatal anamnesis, clinical course ofintrauterine enteroviral infection.Materials and methods. Clinical and laboratory examination of 834 mothers and their sick neonates with suspected intrauterine infections in the first 3 days of life was conducted. The subject of the study was a group of 224 infants with congenital viral infections. Intrauterineenteroviral infection was verified in 108 cases. General clinical and special methods of research were used. Specific diagnosis of intrauterine enteroviral infection was made using the cultural method, polymerase chain reaction and the modified reaction of complement binding withthe use of poly- and monovalent enteroviral diagnostic sera on the enzyme-immunoassay analyzer. Material for examination of children and mothers was the blood taken from the umbilical and peripheral veins and feces. Statistical processing was carried out with the computation of parametric and nonparametric criteria. The results of the research. In 224 children with congenital viral infections during verification of an etiological pathogen, prenatal enteroviral infections was diagnosed in 108 cases (48.2%), including 54 (50,0%) mature and 54 (50.0%) premature ones. In most casesenteroviruses were verified in association with other intrauterine enteroviral infection pathogens (mixed forms) in 91 (84,3%): in association with respiratory vi ruses in 33 (36,3%), with the causative agents of TORCH-group in 27 (29,7%), with bacterial pathogens in 31 (34,0%) and in the form of monoinfection in 17 (15,7%) neonates. On the basis of laboratory examinations the role of various serologicaltypes of enteroviruses in the structure of congenital viral infections in neonates was identified for the first time, with serotypes of entero 68-71 – in 67 (62,0%), echo – in 32 (29,6%) and Coxsackie B – in 15 (13,9%) predominating.The most severe course of infection was observed in premature infants. The most severe course of intrauterine enteroviral infection with the fatal outcome was observed in neonates in case of association of Coxsackie B6 with bacterial pathogens and influenza virus.Summary. On the basis of specific diagnostic methods, the role of various enteroviruses serotypes in the pathology of the fetus and neonates was identified. The most common perinatal risk factors for the development of intrauterine enteroviral infection in neonates are the presence of acute respiratory viral infections in pregnant women,abortion, premature labor, anemia, placental dysfunction, preeclampsia. The fact that enteroviruses cause visceral forms of lesions (serotypes of Coxacques B and ECHO) as well as subclinical forms of infection (serotypes of entero-68-71) in newborns was proved. Increased frequency (48,2%) of enteroviral isolation in neonate patients withintrauterine enteroviral infections, peculiarities of the severe clinical course motivate early diagnosis in case of visceral and generalized forms of intrauterine infections.

Experimental Substantiation of New Presentation Form of Cholera Diagnostic Sera

Objective of the study is experimental substantiation of possibility to produce new presentation form of cholera diagnostic sera – lyophilizate in bottles. Materials and methods. Cholera diagnostic sera. Liophylization was performed in Martin Christ Epsilon 2-6D. Residual moisture of dry sera was determined using humidity meter Sartorius MA 150. Solubility of experimental samples was assessed visually. pH was evaluated by potentiometry with the help of PK SevenExcellence-475 Mettler Toledo device, measuring pH/ORP/Ion content/conductivity/concentration. Specific activity and specificity was analyzed through expanded agglutination reaction, using relevant control strains of cholera vibrio. Results and conclusions. Experimentally justified is the possibility to lyophilize cholera diagnostic sera in flasks. Specified are the optimum parameters of lyophilization. It is determined that lyophilization does not result in deterioration of properties, as judging by the quality indicators they meet the requirements of regulatory documentation. Using accelerated aging test, it is demonstrated that the new presentation form of cholera diagnostic sera – lyophilizate in flasks – maintains their specific activity within five years term (the set out shelf life), which fully conform to normative standards.

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.

EPIDEMIC RISE IN THE INCIDENCE OF INFLUENZA IN ST. PETERSBURG IN 2016

Inf luenza and acute respiratory viral infections (ARVI) account for up to 90% of the total reported infectious morbidity annually in Russia. The economic damage from these infections reaches 86% of the economic losses associated with infectious diseases. As a result of antigenic drift or a shift, new variants of the inf luenza virus, including pandemic strains, appear. In 2009, an inf luenza pandemic arose due to the shifter version of the inf luenza A (H1N1) pdm09 virus. This virus caused epidemic rises of inf luenza in the world in subsequent years. The aim of the study was to describe the epidemic process of inf luenza in January–March 2016 in St. Petersburg. Materials and methods. The incidence of inf luenza and acute respiratory viral infections in St. Petersburg was recorded according to statistical coupons by age groups of the population. Examined nasopharyngeal f lushes received no later than 3–4 days from the onset of the disease, and in case of a lethal outcome — sectional material (bronchi, trachea, lungs, spleen tissues). The detection of RNA of inf luenza viruses was carried out in RT-PCR (Amplicenc Inf luenza virus A/B, Amplicenc Inf luenza virus A/H1-swine-FL, Amplicenc Inf luenza virus A типа-FL, «Interlabservice», Moscow). Isolation of inf luenza viruses from the clinical material was carried out on culture by the MDSC cell. Isolates were typed in RTGA with diagnostic sera to reference and epidemic inf luenza viruses. Isolation of inf luenza viruses from the clinical material was carried out on culture by the MDSC cell. Isolates were typed in RTGA with diagnostic sera to reference and epidemic inf luenza viruses. Results. In 2016, the incidence of inf luenza in St. Petersburg was 81.38 per 100 000 population and was 4.1 times higher than the incidence rate in 2015. The epidemic rise in the incidence of inf luenza and ARVI began with the 3rd calendar week, lasted 6 weeks, with a peak incidence at the 5th calendar week. In total, 398 675 people (7.6% of the population) were ill during this period, including adults — 184 658 (4%), children 0–14 years old — 214 017 or 31.5% of this age group. The greatest number of cases among children was registered in the group of 3–6 years — 87 766 (41.0%). Among the ill children, 86.2% attended pre-school children’s institutions (kindergartens). During the epidemic recovery in 2016, inf luenza viruses were detected in 297 people (46.2%): the inf luenza A (H1N1) virus pdm09 in 97.1% of cases, A (H3N2) — 0.6%, B — 2.3%. There were 102 deaths from complications after the inf luenza, including 101 cases of inf luenza A (H1N1) and A (H3N2) inf luenza A (H1N1) pdm09 confirmed by PCR. Among the dead 101 adults (including 2 pregnant women) and one child. All of them were not vaccinated against the f lu earlier. All deceased adults suffered from chronic physical illnesses.

DOT-IMMUNOASSAY USING GOLD NANOPARTICLE MARKED COLLOID GOLD FOR THE DETECTION OF BOTULINIC TOXIN IN CLINICAL MATERIAL AND FOOD PRODUCTS

Aim. Construction of test-systems for dot-immunoassay using colloid gold nanoparticles as a marker of specific antibodies for the detection of botulinic toxin in clinical material and food products. Materials and methods. 20 nm gold nanoparticles were used as a marker of specific antibodies. IgGs were isolated from polyvalent diagnostic sera against type А, В, С, E, F botulin toxins produced by SPC Allergen (Stavropol) with 5000 - 10 000 ME activity. Botulin toxin in clinical material (blood sera) from 3 patients with established botulism clinical diagnosis as well as food product (home-made mushroom soup solyanka) was determined by dot immunoassay on nitrocellulose membrane. Results. Botulin toxin was detected in all the studied samples (blood sera from 3 patients and the soup) that was registered in the patient No. 1 at the 1:2112 dilution of fhe studied sample, in patient No. 2 - 1:32, in patients No. 3 - 1:1056, in the food product - 1:8. Botulin toxin was not detected in the negative control (pure cultures of the dysentery cauzative agents and intestine yersinosis, blood sera of the patient with All and a healthy individual as well as canned beans in tomato sauce and canned green peas). Conclusion. A highly sensitive specific test-system was developed for dot-immunoassay based on the commercial anti-botulin antibodies labelled with colloid gold particles that allows to detect botulin toxins within 2 hours in the sample volume of 1 - 2 microlites .

Application of Imunofan, Polioksidoniy, Ronkoleykin, Salmozan and Fosprenil for Stimulating the Humoral Immune Response in Rabbits Immunized against Enteroviruses

Enterprise of viral and bacterial Preparations of Chumakov Institute of poliomyelitis and viral encephalitides, Moscow Abstract Selective and inconstant adjuvant activities of the agents Imunofan, Polioksidoniy, Roncoleykin, Salmozan and Fosprenil were determined. Those agents were used as an additional stimulant of the immune response in rabbit immunized with enteroviruses to get the polyclonal diagnostic sera for neutralization.