General features, pathogenesis, and laboratory diagnostics of autoimmune encephalitis.

Autoimmune encephalitis (AE) is a group of inflammatory conditions that can associate with the presence of antibodies directed to neuronal intracellular, or cell surface antigens. These disorders are increasingly recognized as an important differential diagnosis of infectious encephalitis and of other common neuropsychiatric conditions. Autoantibody diagnostics plays a pivotal role for accurate diagnosis of AE, which is of utmost importance for the prompt recognition and early treatment. Several AE subgroups can be identified, either according to the prominent clinical phenotype, presence of a concomitant tumor, or type of neuronal autoantibody, and recent diagnostic criteria have provided important insights into AE classification. Antibodies to neuronal intracellular antigens typically associate with paraneoplastic neurological syndromes and poor prognosis, whereas antibodies to synaptic/neuronal cell surface antigens characterize many AE subtypes that associate with tumors less frequently, and that are often immunotherapy-responsive. In addition to the general features of AE, we review current knowledge on the pathogenic mechanisms underlying these disorders, focusing mainly on the potential role of neuronal antibodies in the most frequent conditions, and highlight current theories and controversies. Then, we dissect the crucial aspects of the laboratory diagnostics of neuronal antibodies, which represents an actual challenge for both pathologists and neurologists. Indeed, this diagnostics entails technical difficulties, along with particularly interesting novel features and pitfalls. The novelties especially apply to the wide range of assays used, including specific tissue-based and cell-based assays. These assays can be developed in-house, usually in specialized laboratories, or are commercially available. They are widely used in clinical immunology and in clinical chemistry laboratories, with relevant differences in analytic performance. Indeed, several data indicate that in-house assays could perform better than commercial kits, notwithstanding that the former are based on non-standardized protocols. Moreover, they need expertise and laboratory facilities usually unavailable in clinical chemistry laboratories. Together with the data of the literature, we critically evaluate the analytical performance of the in-house vs commercial kit-based approach. Finally, we propose an algorithm aimed at integrating the present strategies of the laboratory diagnostics in AE for the best clinical management of patients with these disorders.

Differential diagnostics of food allergy as based on provocation tests and laboratory diagnostic assays.

Due to the scale of the phenomenon, food allergy constitute a significant health problem and significantly impair the quality of life of patients. Differential diagnostics, including skin tests, sIgE detection tests, basophil and mast cell activation tests as well as double-blind placebo-controlled food challenge tests, is the gold standard in the diagnosis of food allergy. Recently, increasing attention has been paid to the potential use of nasal provocation test in the diagnosis of food allergy. Allergen dose, protocol standardization, assessment of subjective complaints and objectivization of test results are important factors determining the applicability of provocation tests.

[Pathogenetic and chemical-laboratory aspects of age-associated vascular diseases of older and senile persons (review).]

Cardiovascular diseases, including atheroscleroscoronary and cerebral arteries, are an important medical and social problem of the population, especially among the elderly and seniors. The proportion of older persons is about 80% among deaths from complications of atherosclerosis and over 60% among all detected cases of myocardial infarction in the Russian Federation. Age changes in the body negatively affect the course of the disease. Analysis of the scientific literature found age-related morphofunctional metamorphoses of the vascular wall, blood forms, lipid and carbohydrate metabolism, immune system, redox balance, microbiota. These changes exacerbate atherogenesis, as atherosclerosis is found to be a multi-factor disease with a wide range of causal relationships. Research demonstrates the possibility of developing measures that can affect different pathogenetic links of the disease. A study of transendothelial transport of lipoproteins, the role of different macrophage populations on the course of atherosclerosis, seems promising. Preventive measures aimed at preventing, early detection of both atherosclerosis itself and its complications, as well as expanding the spectrum of targeted therapy, can significantly improve the quality of life of the older generation.

MODERN DIAGNOSTIC CRITERION FOR ESTABLISHING THE REGIONAL ORIGIN OF BLOOD IN SEXUAL CRIMES.

OBJECTIVE: The aim: Determination of regional blood origin in cases of sexual violence, establishing the possibility of using prostaglandin F2alpha as a marker of blood of menstrual origin. PATIENTS AND METHODS: Materials and methods: The material for the study were samples of vaginal fluid, menstrual blood and capillary blood from females, the age distribution of women was carried out according to the gynecological classification according to the age periods of women's lives depending on the functional state of their reproductive system: the first group - women at the age of 18-29, the second group - women at the age of 30-45. RESULTS: Results: Among objects of biological origin, fluids, especially blood, occupy an important place. The content of PGF2α has age fluctuations: its content is higher by 6-12% in women aged 30-45 than in women 18-29 years old. PGF2α levels above 13.1 ng / mg of dry tissue are a reliable sign of blood of menstrual origin, which is very important in determining the regional origin of blood in forensic examinations for sexual violence / sexual crimes. CONCLUSION: Conclusions: In cases of sexual violence against women, in addition to recording external harm, attention should also be paid to the examination of objects of biological origin, identification by species, sex, regional, organ or cell.

Isolation and identification of listeria in clinical material.

Results from research on isolation, identification, and study of biological properties of L. monocytogenes clinical isolates and Listeria spp test strains are presented. Peculiarities of modern research methods for indicating and identifying pathogenic listeria to improve the quality of laboratory studies of clinical material are studied. The culture method provides reliable results of microbiological analyses upon detecting Listeria spp. The presented list and algorithm of the laboratory diagnostic methods can be used as a basis for elaborating regulatory documents for carrying out microbiological research on any biological material for the presence of bacteria of the genus Listeria spp. and L. monocytogenes species in it.

Educational video for training specialists in clinical laboratory diagnostics.

The adaptation of educational programs under restrictions during the SARS-CoV-2 coronavirus pandemic and after it is based not only on the widespread use of video lectures, but also on the introduction of a number of new educational technologies. Hybrid learning will become the cornerstone of future educational technologies in clinical laboratory diagnostics and will contribute to the creation of virtual practical classes with examples of analysis of laboratory testing results based on case histories (Case Technologies). The key aspect in the implementation of video cases into the educational process is the development of video materials. The use of educational video cases developed by Vector-Best in the process of training specialists in clinical laboratory diagnostics during the cycles «Retraining¼ and «General Improvement¼ at the Department of Clinical Laboratory Diagnostics of the Saratov State Medical University named after V. I. Razumovsky of the Ministry of Health of Russia was a convenient format and received positive assessment of cadets. Currently, the «IVD gallery¼ section has appeared on the FLM website and the placement of an additional educational resource - a library of educational video cases.

[The comparative analysis of professional standards of specialists with higher and secondary medical professional education].

In 2020, the Mintrud of Russia approved a number of professional standards for specialists with secondary professional medical education. The implementation of professional standards is called to actualize outdated normative base concerning functions of medical workers, including assurance of separation of labor functions and actions of physicians and medical nurses, facilitation of development of job descriptions, and minimizing number of conflicts that occur during process of work activities at personnel functions crossing. The medical organization, focusing on requirements established by professional standards, can more competently develop personnel policy, make timely changes in staff list, establish progressive remuneration system. In this regard, it is useful to learn to what extent approved professional standards facilitate solution of practical problems of medical organizations.The article presents results of comparative analysis of three pairs of professional standards for paramedical personnel and specialists with higher education in comparable specialties. Certain contradictions and inaccuracies were esnablished too.

SARS-CoV-2 diagnostics in the virology laboratory of a University Hospital in Rome during the lockdown period.

Italy was one of the most affected nations by coronavirus disease 2019 outside China. The infections, initially limited to Northern Italy, spread to all other Italian regions. This study aims to provide a snapshot of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) epidemiology based on a single-center laboratory experience in Rome. The study retrospectively included 6565 subjects tested for SARS-CoV-2 at the Laboratory of Virology of Sapienza University Hospital in Rome from 6 March to 4 May. A total of 9995 clinical specimens were analyzed, including nasopharyngeal swabs, bronchoalveolar lavage fluids, gargle lavages, stools, pleural fluids, and cerebrospinal fluids. Positivity to SARS-CoV-2 was detected in 8% (527/6565) of individuals, increased with age, and was higher in male patients (P < .001). The number of new confirmed cases reached a peak on 18 March and then decreased. The virus was detected in respiratory samples, in stool and in pleural fluids, while none of gargle lavage or cerebrospinal fluid samples gave a positive result. This analysis allowed to gather comprehensive information on SARS-CoV-2 epidemiology in our area, highlighting positivity variations over time and in different sex and age group and the need for a continuous surveillance of the infection, mostly because the pandemic evolution remains unknown.

COVID-19: advance in laboratory diagnostic strategy and technology.

SARS-CoV-2 is one of the beta-coronaviruses with the spike protein. It invades host cells by binding to angiotensin converting enzyme 2 (ACE2). This newly discovered virus can result in excessive inflammation and immune pathological damage, as shown by a decreased number of peripheral lymphocytes, increased levels of cytokines, and damages of lung, heart, liver, kidney, and other organs. Effective therapeutic modalities such as new antiviral drugs and vaccines against this emerging virus need to be thoroughly studied and developed. However, so far the only recognized but mild progress in this area is the screening of old drugs for new uses. Therefore, rapid and accurate laboratory SARS-CoV-2 testing approaches are the important basis of identification and blockage of COVID-19 transmission. For COVID-19 patients with different clinical classifications (mild, common, severe, and critically severe), dynamic monitoring of functional indicators of susceptible and vital organs is an important strategy for evaluating therapeutic efficacy and prognosis. In this review, we summarized SARS-CoV-2 laboratory diagnostic schemes, pathophysiological indices of tissues and organs of COVID-19 patients, and laboratory diagnostic strategies for distinct disease stages. Further, we discussed the importance of hierarchical management and dynamic observation in SARS-CoV-2 laboratory diagnostics. We then summed up the advance in SARS-CoV-2 testing technology and described the prospect of intelligent medicine in the prevention of infectious disease outbreaks.

Comparison of nine different commercially available molecular assays for detection of SARS-CoV-2 RNA.

To face the COVID-19 pandemic, the need for fast and reliable diagnostic assays for the detection of SARS-CoV-2 is immense. We describe our laboratory experiences evaluating nine commercially available real-time RT-PCR assays. We found that assays differed considerably in performance and validation before routine use is mandatory.

Hemoglobinopathy gone astray-three novel forms of α-thalassemia in Norwegian patients characterized by quantitative real-time PCR and DNA sequencing.

α-thalassemia is one of the most common monogenic diseases worldwide and is caused by reduced or absent synthesis of α-globin chains, most commonly due to deletions of one or more of the α-globin genes. α-thalassemia occurs with high frequency in tropical and subtropical regions of the world and are very rarely found in the indigenous Scandinavian population. Here, we describe four rare forms of α-thalassemia out of which three are novel, found in together 20 patients of Norwegian origin. The study patients were diagnosed during routine hemoglobinopathy evaluation carried out at the Department of Medical Biochemistry, Oslo University Hospital, Norway. The patients were selected for their thalassemic phenotype, despite Norway as country of origin. All samples went through standard hemoglobinopathy evaluation. DNA sequencing and copy number variation (CNV) analysis using quantitative real-time polymerase chain reaction (qPCR) was applied to detect sequence variants and uncommon deletions in the α-globin gene cluster, respectively. Deletion breakpoints were characterized using gap-PCR and DNA sequencing. DNA sequencing revealed a single nucleotide deletion in exon 3 of the HBA2 gene (NM_000517.4(HBA2):c.345del) and a novel deletion of 20 nucleotides in exon 2 of the HBA2 gene (NM_000517.4(HBA2):c.142_161del). qPCR CNV analysis detected two novel large deletions in the α-globin gene cluster, -(NOR) deletion covering both α-globin genes and (αα)Aurora Borealis affecting the regulatory region, leaving the downstream α-globin genes intact. Even though inherited globin gene disorders are extremely rare in indigenous Scandinavians, the possibility of a carrier state should not be ignored.

[Antinuclear antibodies : Practical diagnostic recommendations for dermatologists]. / Antinukleäre Antikörper : Praktische Empfehlungen zur Diagnostik durch den Dermatologen.

Antinuclear antibodies (ANA) are a common diagnostic finding in patients with systemic autoimmune diseases. In patients with typical clinical symptoms, the determination of ANA is of both diagnostic and prognostic importance. However, if ANA were determined due to unspecific symptoms, the interpretation of ANA findings can be problematic. In these cases, misjudgements with severe consequences for the patients are possible. Many systemic autoimmune diseases have prominent early skin involvement and the dermatologist can be the first physician that such a patient sees. Therefore, knowledge of ANA diagnostics is important for dermatologists. Basic principles of autoantibody diagnostics, guidance for the interpretation of laboratory results and new developments are discussed in this overview.

Current state of chikungunya fever laboratory diagnosis (review of literature).

The article is about methods of chikungunya fever laboratory diagnosis. An algorithm for the study of biological material for the presence of antibodies against chikungunya virus and virus antigens is presented. The overview describes the information about commercial immunodiagnostic and genodiagnostic kits and their detailed specifications. The information presented in the review will be useful for doctors of clinical laboratory diagnostics to choose a method and an acceptable test system for laboratory confirmation of Chikungunya fever diagnosis, as well as differential diagnosis with other fevers, which have similar symptoms, common geographical distribution and carriers of infection.

Diagnostic value of microbiome biomarkers of the periodonite microbiome in patients with the association of chronic periodontitis and diabetes mellitus type 2.

The place of high-tech methods of molecular biology in clinical laboratory diagnostics of various diseases and the development of a system of biomarkers as an important component of diagnostic research is currently attracting the closest attention of the scientific community. In this paper, an attempt is made to use high-tech metagenomic analysis to solve problems that arise due to the high frequency of association of periodontal diseases with systemic pathology, in particular, with type 2 diabetes mellitus. The aim of the study was to determine the taxonomic and metabolic features of the microbiome of periodontal tissues in periodontal diseases associated with type 2 diabetes mellitus, as a model of the ratio of local and systemic effects of periodontal pathogenic bacteria. The study included 16S shotgun sequencing of bacterial DNA as part of biological material from periodontal pockets/dentoalveolar furrows of 46 people - 15 patients with chronic periodontitis associated with type 2 diabetes mellitus, 15 patients with chronic periodontitis unrelated to systemic pathology, as well as 16 healthy people in the control group, followed by bioinformatic processing of the data obtained. The obtained data allowed us to establish the taxonomic features of the periodontal microbiome in the association of chronic periodontitis with type 2 diabetes mellitus, which included the predominance of representatives of the families Prevotellaceae and Spirochaetaceae in its composition. The features of metabolic processes in periodontal tissues with the participation of the microbiome were also revealed, which consisted in an increase in the exchange of cysteine and methionine against the background of a decrease in the metabolism of pyrimidine, methane, sphingolipids, and the synthesis of fatty acids, which are of diagnostic value in assessing the condition of patients with type 2 diabetes mellitus.

Development of test kit for detection of specific IgM to SARS-CoV-2 by immune blotting in the «Line blot¼ format.

Test kit for detection of specific IgM to SARS-CoV-2 by immune blotting in the «Line blot¼ format has been developed. A preliminary study of diagnostic effectivity on clinical samples of blood serum from patients with COVID-19 and healthy donors showed its high sensitivity and specificity. The new test kit allows to detect IgM to all four structural antigens of SARS-CoV-2 and can be used as a confirmatory test to verify indeterminant screening results in laboratory etiological diagnosis of COVID-19.

Rapid establishment of laboratory diagnostics for the novel coronavirus SARS-CoV-2 in Bavaria, Germany, February 2020.

The need for timely establishment of diagnostic assays arose when Germany was confronted with the first travel-associated outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Europe. We describe our laboratory experiences during a large contact tracing investigation, comparing previously published real-time RT-PCR assays in different PCR systems and a commercial kit. We found that assay performance using the same primers and probes with different PCR systems varied and the commercial kit performed well.

[Laboratory diagnostics in medicine].

The development of clinical laboratory diagnostics is in line with the evidence-based medicine, which requires that clinical decisions have to be based on diagnostic methods with proven informativity. This creates a request for the scientific validity of the use of laboratory researches and application of probabilistic interpretation tools corresponding to the tasks. The concept of indefiniteness (analytical, biological and clinical) is at the heart of interpretation of laboratory results. The inclusion of laboratory research in clinical guidelines, the choice and appointment of this research to the patient should not be made from the position of ideas about increasing or decreasing the laboratory index in the disease, but on the basis of its scientifically proven characteristics as a laboratory biomarker sensitivity, specificity, predictive value, as well as the relationship with certain clinical events, outcomes, risks. These characteristics are probabilistic and can be defined.

[Diagnosis and prediction of the course of primary open-angle glaucoma by the level of local cytokines]. / Diagnostika i prognozirovanie pervichnoi otkrytougol'noi glaukomy po urovnyu mestnykh tsitokinov.

Glaucoma is the leading cause of irreversible blindness in all countries and is expected to increase in patients with this pathology in the coming years. Currently, the development of the disease is associated with a change in the interleukin profile of the lacrimal fluid, but the diagnostic significance of interleukins has not been sufficiently studied. PURPOSE: To determine the diagnostic significance of interleukins at the local level to improve the diagnosis and prognosis of glaucoma. MATERIAL AND METHODS: The study examined the content of interleukins in the lacrimal fluid of 109 patients with stage II glaucoma and 52 healthy individuals by sandwich-variant of enzyme-linked immunosorbent assay (ELISA) with calculation of the information content of immune parameters. RESULTS: The study has established high diagnostic significance for the verification of glaucoma IL-2, IL-17, IL-8, which is proposed to be used in laboratory diagnosis and prognosis of the disease. CONCLUSION: The created regression model provides high-accuracy prediction of the course of glaucoma progression with increasing IL-2, IL-17, IL-8.

[Competence-oriented exam in clinical laboratory diagnostics for students studying in the specialty «Medical Biochemistry¼.]

In connection with the introduction of annexes and changes to the Federal State Educational Standard of Higher Education in the specialty 30.05.01 «Medical Biochemistry¼, a need arose to improve the assessment tool of general professional and professional competencies formation in the discipline - clinical laboratory diagnostics. A competence-oriented exam (COE) in the studied discipline is developed and implemented. Admission to the exam is the development of practical skills by students in the discipline modules, portfolio design and posting the results on the "Moodle" platform. COE includes two stages: testing to assess the level of theoretical knowledge deepening and solving a situational problem to assess the skills and abilities, interpret the results of laboratory tests and prepare a clinical and laboratory report. Situational tasks and checklists have been developed that make it possible to conduct an objective and comprehensive assessment of the competencies formed by students during the training process, which allows the graduate completed the program to solve professional problems.

Clinical significance of laboratory diagnostics of coxyellosis in children.

The urgency of the problem of coxyellosis in children is determined by the endemic nature of this pathology for a number of regions of Russia. The purpose of the study: to evaluate the results of diagnosis of coxyellosis in children using the methods of complement binding reaction (RSC), enzyme immunoassay (ELISA), and polymerase chain reaction (PCR). Retrospective analysis of the survey on Coxiella in 3 groups of children aged 7 to 17 years: group 1 (n=30) method RSK; group 2 (n=34) - by ELISA; group 3 (n=35) - PCR, were hospitalized in GBUZ «Regional clinical infectious hospital named. A. M. Nicholi¼ Astrakhan in the period from January 2010 to January 2020. The most informative methods of diagnosis of coxyellosis in children during the first 7 days from the onset of the disease is the PCR reaction (specificity-94%, sensitivity-91%), after the 7th day of the disease ELISA (specificity -91%, sensitivity - 94%).The sensitivity of the RSC method is 70%, the specificity is 87%.