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The high demand for SARS-CoV-2 tests but limited supply to South African laboratories early in the COVID-19 pandemic resulted in a heterogenous diagnostic footprint of open and closed molecular testing platforms being implemented. Ongoing monitoring of the performance of these multiple and varied systems required novel approaches, especially during the circulation of variants. The National Health Laboratory Service centrally collected cycle threshold (Ct) values from 1,497,669 test results reported from 6 commonly used PCR assays in 36 months, and visually monitored changes in their median Ct within a 28-day centered moving average for each assays' gene targets. This continuous quality monitoring rapidly identified delayed hybridization of RdRp in the Allplex™ SARS-CoV-2 assay due to the Delta (B.1.617.2) variant; S-gene target failure in the TaqPath™ COVID-19 assay due to B.1.1.7 (Alpha) and the B.1.1.529 (Omicron); and recently E-gene delayed hybridization in the Xpert® Xpress SARS-CoV-2 due to XBB.1.5. This near "real-time" monitoring helped inform the need for sequencing and the importance of multiplex molecular nucleic acid amplification technology designs used in diagnostics for patient care. This continuous quality monitoring approach at the granularity of Ct values should be included in ongoing surveillance and with application to other disease use cases that rely on molecular diagnostics. [ABSTRACT FROM AUTHOR]

Background and Aims: The main objective of this study is to establish the characteristics of blood laboratory parameters in hospitalized coronavirus disease 2019 (COVID‐19) Ukrainian patients and the significance of the above‐mentioned parameters for predicting the course of the disease. Methods: Hematocytological, biochemical, and hemostasis methods of research have been used. Groups of patients with different forms of the coronavirus disease course have been analyzed (lethality ‐ recovery, recovery with a severe and mild course). Results and Conclusion: Age is one of the risk factors for COVID‐19 mortality. Absolute values of neutrophils, neutrophil‐lymphocyte ratio (NLR), systemic inflammation index, d‐dimer, C‐reactive protein, and soluble fibrin complex can be used by clinicians to effectively differentiate between two possible outcomes (lethality vs. recovery). A higher number of stab leukocytes, d‐NLR, and platelets concentration have been recorded in patients with severe COVID‐19 cases, compared to mild ones. The risk of adverse COVID‐19 outcome (lethality) is significantly linked with d‐dimer and NLR (odds ratio 1.42). The risk of a severe course of the disease was significantly associated with the count of leukocytes (odds ratio 4.96). [ABSTRACT FROM AUTHOR]

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Relevance . Despite the large list of biological markers of cardiovascular diseases, not all have evidence-b ased effectiveness and independent prognostic value. Laboratory diagnostics of serum cardiospecific auto-antibodies for the diagnosis of myocyte cell damage has several potential advantages compared to the evaluation of traditional methods. These include the analysis of natural globulins to troponin I (cTnI), to alpha-a ctin 1 (ACTC1), to the heavy chain of beta-myosin 7B (MUN7B), which are based on a self-sustaining immune response to the myocardium’s own auto-antigens, which leads to damage to the cells expressing them. Purpose: To determine the diagnostic and practical value of quantitative indicators for the autoantibody complex to cardiomyocyte proteins to troponin I, to alpha-a ctin 1 and to the heavy chain of beta-myosin 7B in patients with cardiac pathology. Materials and Methods. The study of auto-antibodies to cTnI, ACTC1 and MUN7B in blood serum using laboratory enzyme immunoassay was carried out in patients with cardiac pathology undergoing inpatient treatment at the Regional Clinical Cardiology Dispensary in Stavropol. Additionally, an instrumental and laboratory examination was carried out in accordance with the clinical recommendations developed by the Association of Cardiovascular Surgeons, the Cardiological Society of Russia and approved by the Scientific and Practical Council of the Ministry of Health of the Russian Federation. The work was examined and approved by the Ethics Committee of the North Caucasus Federal University. Results and Discussion . Changes in the level of autoantibodies to cTnI, ACTC1 and MUN7B proteins in blood serum were statistically significant (p 0.01 v. s. p 0.01). A persistent increase in the level of auto-antibodies to cTnI by 2.36 ng/ml (694.11 %), to ACTC1 by 3.6 ng/ml (141.73 %) and to MUN7B by 1.74 ng/ml (119.17 %) was found in individuals with confirmed cardiac pathology, when other criteria for laboratory analysis were within acceptable values, which determine their diagnostic and evidentiary effectiveness. Conclusion . The results of the study showed the relationship of changes in the activity of cardiospecific auto-A T to cardiomyocyte proteins (Anti-cTnI, Anti ACTC1, Anti-M YH7B) in patients with cardiac pathologies, indicating not only systemic membrane disorders (membranopathies), but also serve as convincing evidence of direct chemical changes in cardiomyocytes. A correlation has also been established between cardiomarkers of necrosis and ischemia and autoimmune globulins Anti-cTnI, Anti ACTC1, Anti-MYH7B, that confirms diagnostic and practical value of this laboratory analysis.

Introduction. The World Health Organization estimates that as of 2019, more than 296 million people were living with chronic hepatitis B virus (HBV) infection. The prevalence of HBsAg-negative, occult form of the disease in blood donors varies depending on the region of the world and the sensitivity of the methods of analysis used. Considering that the genetic diversity of viruses demonstrates space and time variations and taking into account that the genetic profile of isolates in key groups, which may turn into a source of the pathogen spread, is important for forecasting of the epidemiological situation, the attention should be given to identification of HBV genotypes currently circulating among regular blood donors in regions of the Russian Federation. The aim of this work was molecular and genetic characterization of HBV genomes identified in HBsAg-negative blood donors in the Ural Federal District. Materials and methods. The study material was 1400 plasma samples obtained from HBsAg-negative blood donors in Ural Federal District. The study included the testing for HBsAg, anti-HBs IgG and anti-HBcore IgG antibodies, HBV DNA. For all identified HBV DNA containing samples, sequencing and analysis of the nucleotide sequences of the complete HBV genomes were performed. Results. The prevalence of HBV DNA was 4.93%, including 4 (0.28%) cases of false occult hepatitis B. Among anti-HBcore IgG-positive samples, HBV DNA was found in 18.08% of cases, while in persons with detected HBV DNA the anti-HBcore IgG positivity rate was 46.38%. In 8.69% of the isolates, anti-HBs IgG antibodies and viral DNA were detected simultaneously in the absence of anti-HBcore IgG. Based on phylogenetic analysis, HBV subgenotypes distribution in HBsAg-negative blood donors was as follows: D3 53.62%, D2 21.74%, D1 18.84%, C2 5.8%. The high variability in the S, C, P regions of the virus genome in the examined group was shown. In all cases of HBsAg-negative chronic HBV infection identified in blood donors, viral sequences contained at least one amino acid substitution in positions, mutations in which are associated with immune escape. In 3 (4.35%) cases mutations in reverse transcriptase region of P gene that are associated with resistance to the following drugs were identified: lamivudine, telbivudine, entecavir. Mutations in the preCore/Core regions that contribute to the progression of liver disease were also identified. Conclusion. Occult HBsAg-negative chronic HBV infection poses a threat of HBV transmission through transfusion of blood and its components due to the extremely low viral load, which does not allow the virus to be detected using routinely used diagnostic kits. The situation can be exacerbated by the abundance and diversity of virus amino acid substitutions that we have identified, including immune escape mutations, drug resistance mutations, and mutations that contribute to the progression of the disease.

The method of express detection of hypotensive drugs (atenolol, propranolol, nifedipine, enalaprile and verapamil) in urine samples of patients with acute poisonings was developed. The optimum conditions of group identification of hypotensive drugs using TLC were found. Analysis by GC-MS allows to detect both the native substances and their metabolites. The developed method of analysis of antihypertensive drugs can be used for the diagnosis of acute poisoning.

Introduction. The most common etiological agents of ixodid tick-borne borreliosis (ITBB) in Russia are Borrelia garinii, B. afzelii, B. bavariensis. Multilocus sequence typing and multilocus sequence analysis (MLSA) have been used in recent studies for Borrelia species identification. The results of using the MLSA scheme for identification of pathogens causing erythemic forms of ITBB have been presented earlier. The purpose of the study was to explore the possibility of MLSA optimization for laboratory identification of ITBB pathogens. Objectives: comparative analysis of nucleotide sequences of 6 conserved genes (rrs, hbb, fla, groEL, recA, ospA) and the rrfA-rrlB intergenic spacer, which are recommended by the MLSA protocol; identification of the minimum set of genes, the concatenated sequences of which are essential for species identification of Borrelia isolates. Materials and methods. The sequences of the above loci of 23 reference isolates collected from patients with ITBB and assigned, using MLSA, to B. bavariensis were compared with the sequences of similar genes of other Borrelia species available in international databases. The UPGMA method was used to build and analyze dendrograms based on the obtained data. Results. The sequences of ospA gene loci of reference species demonstrated the greatest difference (not less than 8.5%) from the sequences of the above gene in other analyzed species of Borrelia; approximately similar species-related differences (not less than 6.7%) were demonstrated by the comparison of recA gene sequences. The sequences of the identified variants of these two genes in B. bavariensis differed from the sequences of the similar genes in the most closely related species B. garinii. The dendrogram of the concatenated nucleotide sequences of recA and ospA genes demonstrated that it was totally consistent with the results of identification of the isolates based on the MLSA protocol. Conclusion. The optimized approach to MLSA of the B. burgdorferi sensu lato group suggests that species identification should be based on the concatenated analysis of loci of only two genes (recA and ospA) out of 7 loci recommended by the MLSA protocol.

Introduction. As is currently known, the epidemic process in the Kaliningrad Region was mainly associated with the spread of the recombinant form of HIV-1 (CRF03_AB); however, regular HIV importations from other countries and continents has created favorable conditions for emergence and spread of various recombinant forms of the virus. The most complete information on the diversity of recombinant forms in the region is also necessary to understand the structure of drug resistance (DR). The aim of the study was to explore the HIV-1 genetic diversity in the Kaliningrad Region. Materials and methods. We studied 162 blood plasma samples obtained from patients from the Kaliningrad Region, both with confirmed virological failure of antiretroviral therapy (ART) and with newly diagnosed HIV infection. For reverse transcription and amplification of HIV genome fragments, diagnostic AmpliSense HIVResist-Seq. Results and discussion. The various recombinants between subtypes A and B (74%) were predominant in study group: recombinant was between CRF03_AB and subtype A (33.95%) and CRF03_AB-like (13.58%) were the most common. Among the pure subtypes of the virus, subtype A6 (16.67%). The circulation of subtypes B (3.70%) and G (1.23%) was also noted. Ninety-six patients (59.26%) were identified with at least one mutation associated with antiretroviral (ARV) drug resistance. Conclusion. The observed diversity of subtypes and recombinant forms of the virus implies that the new recombinants are actively emerging in the studied region, both between existing recombinant forms and pure subtypes, as well as between pure subtypes.

Infertility is a disease of the reproductive system characterized by lacked clinical pregnancy after 12 or more months of regular unprotected sex. To diagnose the causes and choose the tactics of infertility treatment, a personalized approach to each patient is required, which is beneficial for both the doctor and the patient. A comprehensive examination of patients is important from the point of pharmacoeconomics and clinical efficiency view. Here, a comprehensive diagnostics of the spouses resulting in birth of a healthy child is discussed.

Mitochondrial DNA (mtDNA) damage is closely associated with typical diseases of aging, such as Alzheimer’s or Parkinson’s disease, and other health conditions, such as infertility. This damage manifests in reduced mitochondrial copy number and deletion mutations in mtDNA. Consequently, the analysis of mitochondrial damage by determining the parameters copy number and deletion ratio using quantitative real-time PCR (qPCR) is of interest for clinical diagnostics. To bring the findings from research into laboratory practice, a suitable and reliable process is needed, which must be thoroughly validated. This process includes the software used for the analysis, which must meet extensive regulatory and process requirements. Existing software does not adequately implement the requirements of laboratories and, in particular, does not provide direct support for the calculation of the aforementioned mtDNA parameters. The paper discusses the development of a new software-based analysis workflow that is designed specifically for laboratories to help with the calculation of mtDNA parameters. The software was developed using the User-Centered Design method and is based on the recently introduced prototype, “PlateFlow”. Initial user tests provide positive feedback. In the future, this workflow could form the basis for validations of mitochondrial tests in medical laboratories.

In this work, the authors would like to share their experience in laboratory diagnostics and in the care of individuals affected by SARS-CoV-2 infection and in the early stages of the disease of COVID-19 that this infection can cause. It is based on the results of tests that were carried out in the laboratories of Spadia Lab Ostrava during the gradual introduction and internal verification of laboratory methods in the diagnosis of this infection into routine practice, on the experience of caring for such individuals in the AKIMED Brno outpatient clinic and the knowledge obtained with them from the available literature on this issue. The author’s opinions on the current state of laboratory diagnosis of this infection and the disease caused by it and the possibility of streamlining the care of people affected by this infection and the disease, especially in their early stages, are presented for discussion, even though they are aware that their set of people on whom the data was for the study obtained is small and limiting. The full English version of the article is available here [1]. Keywords: SARS-CoV-2 infection, laboratory diagnostics, COVID-19, humoral and cellular immunity. [ABSTRACT FROM AUTHOR]

Diseases related to chronic persisting inflammation are amongst the largest sources of morbidity and health costs, yet biomarkers for early diagnosis, prognosis, and treatment response are not sufficiently effective. This narrative review discusses how inflammation concepts have evolved from ancient times to the present, and places in perspective the use of blood-based biomarkers to assess chronic inflammatory diseases. From reviews of biomarkers in specific diseases, emerging biomarker classifiers and their clinical utility is discussed. Biomarkers representative of systemic inflammatory response such as C Reactive Protein are distinguished from local tissue inflammation markers such as cell membrane components and molecules involved in matrix degradation. The application of newer methodologies such as gene signatures, non-coding RNA, and artificial intelligence/machine-learning techniques is highlighted. The dearth of novel biomarkers for chronic inflammatory diseases can be ascribed in part to the lack of basic understanding about non-resolving inflammation, and in part by fragmentation of effort whereby individual diseases are studied but their pathophysiologic commonalities and differences are neglected. Finding better blood biomarkers for chronic inflammatory diseases may be best addressed by studying cell and tissue products of local inflammation, augmenting data interpretation by artificial intelligence techniques. [ABSTRACT FROM AUTHOR]

Abstract Background and Aims The main objective of this study is to establish the characteristics of blood laboratory parameters in hospitalized coronavirus disease 2019 (COVID‐19) Ukrainian patients and the significance of the above‐mentioned parameters for predicting the course of the disease. Methods Hematocytological, biochemical, and hemostasis methods of research have been used. Groups of patients with different forms of the coronavirus disease course have been analyzed (lethality ‐ recovery, recovery with a severe and mild course). Results and Conclusion Age is one of the risk factors for COVID‐19 mortality. Absolute values of neutrophils, neutrophil‐lymphocyte ratio (NLR), systemic inflammation index, d‐dimer, C‐reactive protein, and soluble fibrin complex can be used by clinicians to effectively differentiate between two possible outcomes (lethality vs. recovery). A higher number of stab leukocytes, d‐NLR, and platelets concentration have been recorded in patients with severe COVID‐19 cases, compared to mild ones. The risk of adverse COVID‐19 outcome (lethality) is significantly linked with d‐dimer and NLR (odds ratio 1.42). The risk of a severe course of the disease was significantly associated with the count of leukocytes (odds ratio 4.96).

The article presents an outline of the concept of a prototype system allowing for the optimization of inventory management in a diagnostic laboratory on the basis of patients results. The effectiveness of laboratory diagnostics depends largely on the appropriate management of resources and the quality of tests. A functional quality management system is an integral element of every diagnostic laboratory, ensuring reliability and appropriate work standards. This system includes maintaining correct and reliable analytical test results as well as the optimal use of the laboratory equipment’s processing capacity and the appropriate organization of the supply chain—both analytical material and reagents. It is extremely important to avoid situations in which tests cannot be performed due to a lack of reagents, the overloading of analyzers, or improper calibration. Therefore, the accurate prediction of the number of orders is crucial to optimize the laboratory’s operations, both in the short term—for the next few hours and minutes—and in the longer term, even monthly, which will allow for the appropriate planning of reagent stock. As part of the research presented in this article, machine learning methods were used to implement the above functionalities, which allowed for the development of a prototype of a laboratory optimization system using patient test results as a basis.

Background Acute mesenteric ischemia (ami) is diagnosed in 1 case per 5,000–15,000 operations associated with elective abdominal surgical intervention and in 1 case of 1,000 emergency laparotomies associated with the acute abdomen clinical features. Non-occlusive disorders account for 20-30% of all cases of ami. The absence of pathognomonic symptoms and syndromes, as well as specific laboratory markers of ami, determines significant difficulties in the early diagnosis of this disease, especially in critically ill patients.Aim of study To identify the main laboratory markers of acute mesenteric ischemia applicable in the resuscitation and intensive care unit (ricu) based on a systematic literature review.Material and methods A systematic search and selection of publications was carried out in march-april 2021 in accordance with the prisma system checklist (prisma, preferred reporting items for systematic reviews and meta-analyzes). The search used two databases: the cochrane library of systematic reviews (https://www.cochranelibrary.com) and the pubmed bibliographic database (https://pubmed.ncbi.nlm.nih.gov). Key terms in the search parameters were: acute mesenteric ischemia; bowel necrosis predictors; laboratory diagnostics of mesenteric ischemia. Search parameters were limited to the year of publication (not later than 2015). The main results of the review included patients with an established diagnosis of acute mesenteric ischemia, indicating the level of laboratory parameters (ph of mixed venous blood, lactate, d-dimer, white blood count, mpv, i-fabp, serum marker and alpha-glutathione-s-transferase, indicators of the coagulation and hemostasis). Case reports and editorial letters were excluded from the search.Results The threshold value for serum l-lactate ranged from 1.05 mmol/l to 5.6 mmol/l. The sensitivity of serum lactate for the diagnosis of ami ranged from 34.7% (95% ci: 0.82–91.64%) to 89.51% (95.4% ci: 75.12–94.28%), and the specificity ranged from 47.3% (94.7% ci: 26.54–67.98%) to 94.5% (93.2% ci: 75.44–96.85%). The threshold level of d-dimer varied from 0.58 nmol/l to 7.84 nmol/l. The sensitivity of blood d-dimer for the diagnosis of ami was 72.6% (94% ci: 56.1–92.4%) to 99.7% (94% ci: 69.15–100%); the specificity ranged from 69% (95% ci: 7.41–33.63%) to 97.89% (95% ci: 90.42–98.1%). Parameters such as transaminases (ast, alt), mpv, ldh, d-lactate, serum amylase were not included in the review due to the lack of a sufficient number of studies.Conclusion Today, there is no highly specific laboratory marker that could be considered the “gold standard” in the laboratory diagnosis of acute mesenteric ischemia. The most promising in predicting ami are serum levels of l-lactate and d-dimer.

Osteoarthritis is the most common joint disease in animals, which is not always treatable. Both in veterinary and in human medicine, laboratory research methods are used to effectively diagnose this pathology. The purpose of this study was to establish the dynamics of markers of the acute phase of inflammation in the blood using experimental modelling of knee osteoarthritis in rabbits. Artificial reproduction of the pathological process in the knee joints of animals was performed by intra-articular administration of Yellow peel 2*5 ml. The BC-2800Vet automatic haematology analyser (China) was used to calculate the number of white blood cells and their subpopulations in the blood. The content of C-reactive protein in blood serum was found using enzyme-linked immunosorbent assay. The rate of erythrocyte sedimentation in the blood was investigated according to the Panchenkov method. Thus, on Day 7, a sharp increase in the number of white blood cells (2.7 times) and their subpopulations was noted in sick rabbits, which indicates an active inflammatory process of a systemic nature. On Day 14, their number in the blood of sick animals was characterized by a tendency to decrease, but it had not yet reached the values of the control group. On Days 21 and 28, all the indicators under study tended to decrease in sick animals. Specifically, the number of white blood cells and their subpopulations acquired reference values. The content of C-reactive protein significantly increased by 40.8 times in sick animals on Day 7 of the study and decreased by 1.7 times on Day 14 compared to its values at the previous stage. At the same time, on Day 28, this indicator increased by 19.3 times compared to the indicators of the control group of animals. The rate of erythrocyte sedimentation in the blood of sick rabbits significantly increased by 2.3 times compared to the control group. On Day 21, this indicator in the blood of sick rabbits decreased by 1.4 times compared to its values on Day 14 of the experiment. On the other hand, on Day 28, its value increased by 1.7 times compared to the control (4.3 mm/h). The results obtained will contribute to further targeted laboratory diagnostics of knee osteoarthritis in animals, which will ensure the proper effectiveness of therapeutic and preventive measures and improve their overall health

Relevance. The problem of timely and effective diagnosis of COVID-19 remains one of the main problems facing healthcare. In this regard, the task of developing test systems for the etiological diagnosis of COVID-19 remains extremely relevant. Purpose To develop ELISA test system for detection of G specific immunoglobulins to SARS-COV-2 coronavirus by immunoblotting (Line Blot). Methods. Elaboration of techniques for obtaining test components and preliminary assessment of its diagnostic effectivenessin blood serums from COVID-19 patients treated at N. I. Pirogov First Gradsky Hospital, Moscow, and serums from healthy human donors. Results. The study of 104 blood serum samples from COVID-19 patients and 100 blood serum samples from healthy human donors, pre-tested by ELISA IgG to SARS-CoV-2 using «Vitrotest SARS-CoV-2 IgG» test systems (Vitrotest, Ukraine) and «ELISA-SARS-CoV-2-AB-G» (CJSC EKOlab, Russia) showed high diagnostic efficiency of the new test system. Conclusion. The new test system after state registration of the medical device can be recommended as a confirmatory test for the etiological laboratory diagnosis of COVID-19.

Acute kidney injury (AKI) is a common disease, with high morbidity and mortality rates. In this study, we investigated the potential influence of sex and age on laboratory diagnostics and outcomes. It is known that serum creatinine (SCr) has limitations as a laboratory diagnostic parameter for AKI due to its dependence on muscle mass, which may lead to an incorrect or delayed diagnosis for certain patient groups, such as women and the elderly. Overall, 7592 cases with AKI, hospitalized at the University of Leipzig Medical Center (ULMC) between 1st January 2017 and 31st December 2019, were retrospectively analyzed. The diagnosis and staging of AKI were performed according to the Kidney Disease: Improving Global Outcomes (KDIGO) guidelines, based on the level and dynamics of SCr. The impact of sex and age was analyzed by the recalculation of a female to male and an old to young SCr using the CKD-EPI equation. In our study cohort progressive AKI occurred in 19.2% of all cases (n = 1458). Female cases with AKI were underrepresented (40.4%), with a significantly lower first (−3.5 mL/min) and last eGFR (−2.7 mL/min) (p < 0.001). The highest incidence proportion of AKI was found in the [61–81) age group in female (49.5%) and male (52.7%) cases. Females with progressive AKI were underrepresented (p = 0.04). By defining and staging AKI on the basis of relative and absolute changes in the SCr level, it is more difficult for patients with low muscle mass and, thus, a lower baseline SCr to be diagnosed by an absolute SCr increase. AKIN1 and AKIN3 can be diagnosed by a relative or absolute change in SCr. In females, both stages were less frequently detected by an absolute criterion alone (AKIN1 ♀ 20.2%, ♂ 29.5%, p < 0.001; AKIN3 ♀ 13.4%, ♂ 15.2%, p < 0.001). A recalculated SCr for females (as males) and males (as young males) displayed the expected increase in AKI occurrence and severity with age and, in general, in females. Our study illustrates how SCr, as the sole parameter for the diagnosis and staging of AKI, bears the risk of underdiagnosis of patient groups with low muscle mass, such as women and the elderly. A sex- and age-adapted approach might offer advantages. [ABSTRACT FROM AUTHOR]

During early phases of the SARS-CoV-2 epidemic, many research laboratories repurposed their efforts towards developing diagnostic testing that could aid public health surveillance while commercial and public diagnostic laboratories developed capacity and validated large scale testing methods. Simultaneously, the rush to produce point-of-care and diagnostic facility testing resulted in FDA Emergency Use Authorization with scarce and poorly validated clinical samples. Here, we review serologic test results from 186 serum samples collected in early phases of the pandemic (May 2020) from skilled nursing facilities tested with six laboratory-based and two commercially available assays. Serum neutralization titers were used to set cut-off values using positive to negative ratio (P/N) analysis to account for batch effects. We found that laboratory-based receptor binding domain (RBD) binding assays had equivalent or superior sensitivity and specificity compared to commercially available tests. We also determined seroconversion rate and compared with qPCR outcomes. Our work suggests that research laboratory assays can contribute reliable surveillance information and should be considered important adjuncts to commercial laboratory testing facilities during early phases of disease outbreaks. [ABSTRACT FROM AUTHOR]

To define the relationship of SARS-CoV-2 antigen, viral load determined by RT-qPCR, and viral culture detection. Presumptively, viral culture can provide a surrogate measure for infectivity of sampled individuals and thereby inform how and where to most appropriately deploy antigen and nucleic acid amplification-based diagnostic testing modalities. We compared the antigen testing results from three lateral flow and one microfluidics assay to viral culture detection and viral load determination performed in parallel in up to 189 nasopharyngeal swab samples positive for SARS-CoV-2. Sample viral loads, determined by RT-qPCR, were distributed across the range of viral load values observed in our testing population. Antigen tests were predictive of viral culture positivity, with the LumiraDx microfluidics method showing enhanced sensitivity (90%; 95% CI 83–94%) compared with the BD Veritor (74%, 95% CI 65–81%), CareStart (74%, 95% CI 65–81%) and Oscar Corona (74%, 95% CI 65–82%) lateral flow antigen tests. Antigen and viral culture positivity were also highly correlated with sample viral load, with areas under the receiver operator characteristic curves of 0.94 to 0.97 and 0.92, respectively. A viral load threshold of 100 000 copies/mL was 95% sensitive (95% CI, 90–98%) and 72% specific (95% CI, 60–81%) for predicting viral culture positivity. Adjusting for sample dilution inherent in our study design, sensitivities of antigen tests were ≥95% for detection of viral culture positive samples with viral loads >106 genome copies/mL, although specificity of antigen testing was imperfect. Antigen testing results and viral culture were correlated. For culture positive samples, the sensitivity of antigen tests was high at high viral loads that are likely associated with significant infectivity. Therefore, our data provides support for use of antigen testing in ruling out infectivity at the time of sampling. [ABSTRACT FROM AUTHOR]

Introduction The use of laboratory tests to monitor the treatment of patients with chronic osteomyelitis has a lot of clinical relevance. The purpose of the study was to identify laboratory tests for assessing metabolic processes in patients with purulent lesions of the foot bones for monitoring the dynamics of the recovery processes in the course of treatment. Methods The studies were carried out in 36 patients with chronic osteomyelitis of the calcaneus and adjacent bones of the foot of traumatic etiology. All patients were treated with the method of the Ilizarov transosseous osteosynthesis. Biochemical and coagulation parameters were studied. Results It was found that in patients with purulent lesions of the foot bones, the elevated levels of blood serum lactate were most frequently detected before surgery (52.8 %, 95 % CI: 36.6-68.6). The incidence of elevated CRP was 27.7 % (95 % CI: 14.6-43.3) on average, and one third of patients had elevated fibrinogen levels (33.3 %, 95-CI: 19.1-49, 3). During the treatment period, the incidence of elevated lactate levels in patients did not change but the incidence of elevated CRP levels increased significantly which was observed in 77.8 % of patients on the 3rd post-surgery day. By discharge from the hospital, elevated values of CRP and lactate occurred in 11.1 % of patients. It is assumed that the greatest diagnostic significance is the assessment of the level of lactate in the dynamics of the treatment in target patients. Discussion Based on the results of the studies, a laboratory sign was found that indicates the effectiveness of surgical treatment in patients with chronic osteomyelitis of the foot bones using the method of transosseous osteosynthesis. Such a sign is a decrease in the initially elevated concentration of lactate in the blood serum of patients in the postoperative period. The detected sign meets the requirements for its applicability in laboratory tests: 1) this sign is the most common in the target group of patients; 2) the dynamics of changes in the level of lactate correlates with the ongoing treatment; 3) this test is an indicator of a significant physiological process: oxidative metabolic processes in patients with the studied pathology.

The aim of the work was to summarize the experience of the Rostov-on-Don Research Anti-Plague Institute in the organizational and functional response aimed at ensuring the implementation of a complex of on-going and scheduled anti-epidemic (preventive) measures at different territorial levels in the context of COVID-19 pandemic. Stage-wise reassignment of scientific and operational subdivisions of the facility was carried out in a timely manner in a flexible mode for a prompt response to the challenges that arose from community transmission of the new coronavirus infection in the territory of the Russian Federation. It allowed an effective distribution of the forces and assets without damage for specialized functioning in the main areas of scientific and practical activities. Realized was an effective interdepartmental interaction with territorial scientific and practical institutions of the Rospotrebnadzor and specialized medical organizations. Practical, methodological and advisory assistance was provided to personnel of specialized medical institutions within the framework of conversing in-patient facilities into hospitals for the treatment of patients with COVID-19. Priority research to study the level of the humoral and cellular immune response to SARS-CoV-2 among the population of the Rostov Region, the etiological spectrum of the causative agents of community-acquired pneumonia associated with COVID-19, full genome sequencing of SARS-CoV-2 isolates was carried out. Specialists of the task forces reinforced human resources of individual laboratory facilities in Moscow city, the Republic of Crimea and the Rostov Region. Complex objective of effective functioning has been implemented: in an operational mode – participation in anti-epidemic (preventive) measures in the face of changes in the dynamics and intensity of the epidemic manifestations of new coronavirus infection; in scheduled mode – the implementation of activities in the main areas of the scientific work of the organization.

Permanently increased frequency of drug hypersensitivity is observed throughout the world and continues to be of concern to clinicians of either discipline. Diagnostic problems with prevention and treatment of hypersensitivity reactions are due to different pathogenetic mechanisms that may underly similar clinical manifestations. On the other hand, clinical symptoms, response to therapy and the outcome of the disease significantly depend on clinical state of the patient. The phenotypes which are determined by clinical manifestations, and endotypes dependent on the pathogenetic mechanisms of hypersensitivity reactions were specified to the purpose of personalized therapy. Two kinds of phenotypes are described: with immediate (1-6 hours) and delayed (more than 6 hours) onset of the response. Four types of hypersensitivity according to Gell and Coombs with the presence of primary sensitization are discerned between the endotypes. Moreover, interaction of drugs with immune receptors of lymphocytes, according to the Pichler’s pharmaco-immune concept, has been identified, along with quick impact of drugs directly upon receptors of effector cells and signaling cascades without primary sensitization, which was previously termed as “pseudo-allergy”. To optimize therapy and prevent repeated reactions in the patients with drug hypersensitivity, high-quality diagnostics with detection of the causal allergen is required. This strategy currently includes analysis of medical history, physical examination as well as in vivo and in vitro testing. Provocation tests are rarely used, due to the significant risk of severe adverse reactions. Skin tests are a reliable and cost-effective tool, if the case history infers possibility of IgE-dependent reactions. Among nonspecific laboratory markers, the greatest clinical value is shown for determining serum tryptase levels, especially in anaphylactic reactions. In addition to genetic testing, several specific laboratory methods are used, i.e., measuring levels of drug-specific IgE, assessment of antigen-induced activation and proliferation of lymphocytes as well as basophil activation test, which has a number of advantages and allows us to identify a causally significant allergen for the hypersensitivity reactions proceeding by various mechanisms. Due to dysregulation which is a key factor of allergic inflammation, the mechanisms of normalization for impaired immune regulation should be understood, thus offering new strategies for personalized treatment, e.c., modulating the regulatory cell functions, aiming for control of the drug hypersensitivity reactions.

Introduction. The problem of transfusion safety in relation to parenteral viral hepatitis still remains relevant. Viral hepatitis B (HB) remains the most common viral infection transmitted through transfusion procedures. One of the natural phases of chronic hepatitis B (CHB) is occult hepatitis B infection (OBI), characterized by an undetectable HBsAg (regardless of the other serological markers content) in the presence of hepatitis B virus (HBV) DNA in the liver tissue and an extremely low, up to undetectable, level of viral load in the blood. In the Republic of Guinea, as in most countries on the continent, the prevention of HBV transmission through transfusion is still based on HBsAg serological testing of donors only. In this connection, OBI remains as a potential threat to blood transfusion safety. Detection of HBV DNA is a reliable preventive measure against transmission of the virus from donors with HBsAg-negative HBV infection, especially in highly endemic regions. In this regard, the study was conducted to substantiate recommendations for improving blood safety against the background of significant HBV prevalence in the Republic of Guinea. The aim of the work was the evaluation of serological and molecular markers of HBV infection in blood donors in the Republic of Guinea. Material and methods. We examined 250 blood samples obtained from donors living in Conakry, Republic of Guinea. Samples were tested for the presence of serological (surface antigen, HBsAg; antibodies (ABs) to surface (anti-HBs IgG) and core (anti-HBc IgG) antigens) and molecular (DNA) markers of HBV infection. Results and discussion. The overall detection rate of hepatitis B markers was 83.2%; HBsAg was detected in 16.4% of all individuals. The high incidence of HBsAg in men (19.55%) compared to women (8.45%) was shown, the relative risk of HBV infection with the formation of HBsAg-positive chronic hepatitis B in males was also significantly higher. The prevalence of the HBV DNA in the study group was 30.4%, the OBI cases accounted for 15.6%. The prevalence of this form of the disease was shown in donors aged 30–49 years (24.78%), in the group of people younger than 30 years, the incidence was lower (8.73%), and at the age of over 50 years, OBI was not detected. Based on the phylogenetic analysis of 76 virus isolates, it was shown that genotype E prevails in the examined group (85.53%). Cases of pathogen DNA detection occurred in HBsAg-negative blood donors in the presence of anti-HBs IgG (n = 4), as well as in the simultaneous presence of ABs anti-HBs IgG and anti-HBc IgG (n = 7). The viral load exceeded 200 IU/ml in OBI samples. Escape mutations were detected by sequencing in each OBI sample, contributing to the virus escaping from diagnostic based on screening for HBsAg. Conclusion. Assessment of the prevalence viral hepatitis B markers in blood donors, determination of genotypes and clinically significant mutations of virus variants are necessary to ensure safe medical manipulations, control and prevention of the spread of this infectious agent.

Purpose: to study the clinical signs and effectiveness of laboratory diagnostics of fungal corneal ulcer complicated by endophthalmitis aimed at treatment tactics optimization.Materials and methods. 15 patients aged 20–55 with severe monocular fungal ulcer complicated by endophthalmitis were examined by microscopy and culture of corneal scraping on Sabouraud Agar; microscopy and culture of intraocular fluid and corneal fragments, obtained during the surgical treatment. Corneal scrapings from the corneal ulcer and corneal fragments removed during keratoplasty were tested in polymerase chain reaction (PCR) for the presence of deoxyribonucleic acid (DNA) of Candida albicans and total Fungi DNA.Results. Before treatment start, 2.5 hours after the biomaterial was taken, PCR revealed Fungi DNA in all 15 cases. Symptoms of the severe fungal corneal ulcer complicated by endophthalmitis and caused by Aspergillus spp have been identified and described. Conservative therapy and total penetrating keratoplasty (PKP) helped retain the eye in 13 out of 15 patients (86.7 %) and partially restore the visual functions.Conclusion. Rapid laboratory diagnosis of ophthalmomycosis using PCR offers an advantage over the culturing technique: the results are ready within 2.5 hours instead of 10-14 days (culture method); it is highly sensitive and allows identifying the fungi not only in the tissue of the removed cornea, but also in the сorneal scrapings. The effective treatment tactics of fungal corneal ulcer complicated by endophthalmitis combining PKP with pre-operative and long-term postoperative antifungal therapy was proposed.

This article describes the laboratory diagnostics stages of acute erythroid leukemia clinical case. The following research results allow observing the process of disease formation in development.

The aim of this study was to characterize mutations in the hepatitis B virus (HBV) genome associated with HBsAg-negative form of the disease in patients receiving hemodialysis replacement therapy. Materials and methods. We used blood plasma samples obtained from hemodialysis centers in St. Petersburg, Russia – 173 patients and 108 patients from Belgrade, Republic of Serbia. The samples were examined for the presence of serological (HBsAg, antibodies anti-HBs IgG, anti-HBcore IgG) and molecular-genetic (HBV DNA) markers of HBV followed by whole-genome sequencing and determination of clinically significant virus mutations. Results and discussion. Antibodies to hepatitis B were detected in 7.5 % and 11.1 % of patients from St. Petersburg and Belgrade, respectively. HBsAg was identified only in 1.1 % of cases in the group from Russia and in 0.9 % of cases in the group from Serbia. HBV DNA was determined in 2.8 % of the studied samples from both, patients from Saint-Petersburg and Belgrade. Phylogenetic analysis of 9 viral isolates showed that genotype D virus (88.9 %) prevailed as compared to genotype A (11.1 %) in the examined group. Among the samples obtained from patients from St. Petersburg, four belonged to the D2 sub-genotype, one to the D3 genotype. Four samples obtained from Belgrade patients belonged to different sub-genotypes – D1, D2, D3, A2, respectively. When analyzing the nucleotide sequences of the HBV genomes, mutations in the MHR region were detected in all cases, but only in HBsAg-negative isolates, mutations were revealed in the region of 124–147 amino acids, including mutations P120T, R122K, A128V, Q129R, M133I, G145R affecting the recognition of HBsAg by anti-HBs antibodies and associated with the resistance of the virus to the vaccine. The results of this study indicate that transmission of blood-borne viral hepatitis agent in the hemodialysis departments of the Russian Federation and the Republic of Serbia still exists. The prevalence of the latent chronic hepatitis B, coupled with the presence of vaccine escape mutations in all identified cases, indicates the need to pay close attention to the occurrence of the virus mutant variants in hemodialysis centers.

Mitochondrial DNA (mtDNA) damage is closely associated with typical diseases of aging, such as Alzheimer's or Parkinson's disease, and other health conditions, such as infertility. This damage manifests in reduced mitochondrial copy number and deletion mutations in mtDNA. Consequently, the analysis of mitochondrial damage by determining the parameters copy number and deletion ratio using quantitative real-time PCR (qPCR) is of interest for clinical diagnostics. To bring the findings from research into laboratory practice, a suitable and reliable process is needed, which must be thoroughly validated. This process includes the software used for the analysis, which must meet extensive regulatory and process requirements. Existing software does not adequately implement the requirements of laboratories and, in particular, does not provide direct support for the calculation of the aforementioned mtDNA parameters. The paper discusses the development of a new software-based analysis workflow that is designed specifically for laboratories to help with the calculation of mtDNA parameters. The software was developed using the User-Centered Design method and is based on the recently introduced prototype, "PlateFlow". Initial user tests provide positive feedback. In the future, this workflow could form the basis for validations of mitochondrial tests in medical laboratories. [ABSTRACT FROM AUTHOR]

Introduction: Following the introduction of pertussis vaccination during infancy, the age-related demographics of pertussis epidemiology have changed. Methods: To better understand the pertussis burden (defined here as number of cases and/or incidence rate [IR]) among older adults (OA; at least 50 years of age) in Europe, we collected data on the reported number of cases and IR in this population in Denmark, England and Scotland, Finland, Germany, the Netherlands, Norway and Sweden from 2010 to 2020. Additionally, we collected contextual epidemiological information on surveillance systems, case definitions, laboratory diagnostics and vaccination approaches. Results: We observed large heterogeneity in the burden among OA between countries: annual IRs ranged from 0.4 (England, 2010) to 54.5 (Norway, 2011) per 100,000 population; 9% (Denmark, 2010) to 45% (England, 2017) of all reported cases occurred in OA. No clear impact of changes in contextual epidemiological information or common trends between countries could be observed, highlighting the need for standardised pertussis surveillance programmes across Europe. The epidemiological trends observed in OA were similar to those observed in 0–4-year-olds. Conclusion: This analysis showed that B. pertussis continues to circulate among OA in Europe, suggesting that current vaccination strategies are insufficient to decrease the disease burden in all age groups. This may indicate that improved monitoring of pertussis in OA and booster vaccination throughout adulthood are necessary to control the total pertussis burden. Plain Language Summary: Whooping cough is an infectious, vaccine-preventable disease that is primarily serious in unvaccinated infants but can also affect adults (at least 50 years old). While vaccination is well established in children, many countries do not routinely vaccinate older adults. Moreover, whooping cough infections in older adults can be difficult to identify for healthcare professionals because of the atypical and mild nature of symptoms. Consequently, the extent of whooping cough occurrence in this population is underestimated. To better understand the extent of disease occurrence, we studied whooping cough infections in Denmark, England and Scotland, Finland, Germany, the Netherlands, Norway and Sweden from 2010 to 2020. Our study was based on the number of laboratory-confirmed cases reported to relevant institutions. We also assessed whether we could identify links between disease occurrence among older adults and contextual epidemiological information, such as disease monitoring systems, methods used for laboratory confirmation, vaccination schedules and vaccination coverage rates. Our study confirmed that whooping cough affects older adults and disease occurrence follows similar trends to those in 0- to 4-year-old children. Because the contextual epidemiological information differed over time and between countries, we could not establish links with disease occurrence in older adults. These data may provide further evidence to authorities that whooping cough among older adults would be better controlled and its burden more accurately estimated with a reinforced comprehensive approach around vaccination and monitoring. Because adults can also infect children who are not yet fully vaccinated, such an approach might help further control the disease in children. [ABSTRACT FROM AUTHOR]

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Relevance. Bronchial asthma is a serious medical and social problem. To confirm the diagnosis, it is necessary to assess the allergic status, to determine the biological markers of the disease. Of particular relevance is a comprehensive analysis of the results of laboratory examination of patients, including data from hematological, immunological, microbiological studies. Aim. To study some laboratory parameters (hematological, immunological and microbiological), the changes of which are most typical for patients with an established diagnosis of bronchial asthma. Matherials and Methods . A clinical and diagnostic examination of women in the non-production and industrial sectors with an established diagnosis of bronchial asthma, performed during a periodic medical examination, was carried out. Results and Discussion . Laboratory research has shown that hematological parameters are characterized by a reduced content of hemoglobin, erythrocyte indices MCV, MCH, MCHS - markers of hypochromia. The surveyed women had increased indices: eosinophils, immunoglobulin E, allergy index, leukocyte intoxication index. High rates of allergization are found in the production sector in 41,51%, in the non-production sector - in 44,44%, which indicates a serious sensitization of the body. Bacterial contamination in the examined groups is represented by coccal flora, where the most significant microorganisms are Staphylococcus aureus, Staphylococcus spp, Streptococcus pyogenes and Candida albicans. Conclusion . The results of the analysis indicate the need to conduct and study further research on the development of markers of early, prenosological health disorders in the process of screening examination of large groups of the working population.

Aim: To evaluate the epidemic situation on measles and rubella and to carry out the laboratory diagnostics of these infections in the North-West of Russia in the period of elimination.Materials and Methods. In 2016-2019 a total of 534 blood serum samples from patients with measles diagnosis, 210 – from patients with rubella diagnosis and 1251 – from patients with exanthema were studied by ELISA using “VectoMeasles-IgM” and “EIA-Rubella-IgM” test-systems.Results. Measles incidence in NWR varied from 0 to 0.96 per 100,000 in 2016 and 2019, respectively. Among measles patients the proportion of children and adults changed in 2018-2019, adults constituted 65.9%. Patients not vaccinated against measles and those with the unknown vaccination status were mainly involved in epidemic process (85%). High level of patients with revaccination status was revealed in 2018 (18.4%). These patients demonstrated high levels of IgG (>3.0 IU/ml) as well as high percentage of IgG avidity (>94.3%) on 4-7th day of rash onset thus evidencing secondary immune response. Molecular studies of the samples from patients revealed the circulation of D8 and B3 genotypes of measles virus. Strains of D8 and B3 genotypes were represented by 3 and 4 variants, respectively. Rubella incidence did not exceed 0.13 per 100,000. Strains of rubella virus of genotype 1E were detected.Conclusion. The data of annual measles incidence and molecular studies of circulating wild type measles viruses prove the on-going measles elimination process in the NorthWest of Russia. For rubella, the persistent phase of elimination was demonstrated in the region.

The article describes the main biomarkers for determining human adaptation to physical activity. Analyzed modern scientific publications on the criteria of individual tolerance of food products, studied promising directions of personalized correction of the diet. The authors have formed a group of valid (sports-specific) laboratory indicators, it is proposed to pay close attention to the reference values of laboratory equipment that have “their own” norms. A promising direction of laboratory diagnostic work is the formation of reliable and accessible complementary criteria that are at the “junction” of functional and instrumental diagnostic methods. Metabolomics is viewed as a young but highly effective science for detecting highly specific metabolic characteristics of human health. The authors proposed the main directions of scientific research in laboratory diagnostics in sports.

According to the latest data released by UNAIDS, the global number of people living with HIV (PLHIV) in 2021 was 38.4 million, with 1.5 million new HIV infections. In different countries, a significant proportion of these cases occur in the adult fertile population aged 15–49 years. According to UNAIDS, Vietnam had a national HIV prevalence of 0.3% of the total population at the end of 2019, with approximately 230,000 PLHIV. The most effective way to prevent mother-to-child transmission of HIV is ART to reduce maternal viral load. HIV-infected pregnant women should undergo monthly monitoring, especially before the expected date of delivery. The aim of our work was to analyze subtypic structure and drug-resistant variants of HIV in pregnant women in Ho Chi Minh City. The study material was blood plasma samples from HIV-infected pregnant women: 31 women showed virological failure of ART, and 30 women had not previously received therapy. HIV-1 genotyping and mutation detection were performed based on analysis of the nucleotide sequences of the pol gene region. More than 98% of sequences genotyped as HIV-1 sub-subtype CRF01_AE. When assessing the occurrence of drug resistance mutations, genetic resistance to any drug was detected in 74.41% (95% CI: 62.71–85.54%) of patients. These included resistance mutations to protease inhibitors in 60.66% (95% CI: 47.31–72.93%) of patients, to NRTIs in 8.20% (95% CI: 2.72–18.10%), and to NNRTIs in 44.26% (95% CI: 31.55–57.52%). Mutations associated with NRTI (2) and NNRTI (8) resistance as well as PI mutations (12), including minor ones, were identified. The high prevalence of drug resistance mutations found in this study among pregnant women, both in therapeutically naive individuals and in patients with virological failure of ART, indicates that currently used regimens in Vietnam are insufficient to prevent vertical HIV infection.

The high demand for SARS-CoV-2 tests but limited supply to South African laboratories early in the COVID-19 pandemic resulted in a heterogenous diagnostic footprint of open and closed molecular testing platforms being implemented. Ongoing monitoring of the performance of these multiple and varied systems required novel approaches, especially during the circulation of variants. The National Health Laboratory Service centrally collected cycle threshold (Ct) values from 1,497,669 test results reported from 6 commonly used PCR assays in 36 months, and visually monitored changes in their median Ct within a 28-day centered moving average for each assays’ gene targets. This continuous quality monitoring rapidly identified delayed hybridization of RdRp in the Allplex™ SARS-CoV-2 assay due to the Delta (B.1.617.2) variant; S-gene target failure in the TaqPath™ COVID-19 assay due to B.1.1.7 (Alpha) and the B.1.1.529 (Omicron); and recently E-gene delayed hybridization in the Xpert® Xpress SARS-CoV-2 due to XBB.1.5. This near “real-time” monitoring helped inform the need for sequencing and the importance of multiplex molecular nucleic acid amplification technology designs used in diagnostics for patient care. This continuous quality monitoring approach at the granularity of Ct values should be included in ongoing surveillance and with application to other disease use cases that rely on molecular diagnostics.

Invasive candidiasis, including bloodstream infection (candidemia), encompasses the most severe forms of Candida infection. Several species-specific and non-specific serological assays are commercially available to aid in diagnosis. This study compared the performance of five such biomarker assays. Serum samples from 14 patients with proven or probable invasive candidiasis, and from 10 control patients, were included in the analysis. A total of 50 serum samples were tested using C. albicans germ tube antibody (CAGTA) assay (Vircell), C. albicans IgM, C. albicans IgG and Candida mannan assays (Dynamiker Biotechnology). Among these samples, the β-1-3-D-glucan (BDG) assay (Fungitell), a laboratory standard for the diagnosis of invasive candidiasis, was positive in 20 (40%), intermediate in five (10%) and negative in 25 (50%). In cases of proven or probable candidemia, the sensitivity and specificity of the BDG assay was 86% and 80%, respectively; the Candida mannan assay, 14% and 86%; the CAGTA test, 57% and 60%; the C. albicans IgM assay, 71% and 60%; and C. albicans IgG assay 29% and 90%. In 4/8 (50%) cases with multiple serum samples, C. albicans IgM was positive sooner than BDG. Thus, when used as a rule-out test for invasive candidiasis, our data suggest that the C. albicans IgM assay may assist antifungal stewardship (over serum BDG).

Highly active antiretroviral therapy (HAART) is currently a combination of three (less frequently four) antiretroviral drugs; these target pathways involved in various stages of HIV replication in the body. Treatment failure is a problem facing doctors and patients using HAART. The most common cause of therapeutic failure is the development of HIV drug resistance. The emergence of resistance is associated with processes involving mutation occurring in the viral genome under the influence of evolutionary factors. Sequencing reactions were performed using the AmpliSens HIV Resist-Seq. Assembly of consensus sequences from fragments obtained during sequencing was carried out using Unipro UGENE softwar. Isolate genotyping was performed using the MEGA-X software with the Neighbor-joining algorithm. According to the analysis, 72.05% of patients had at least one significant mutation associated with drug resistance for the corresponding viral subtype. HIV-1 A6 remains the predominant HIV-1 genetic variant in Russia's Northwestern Federal District. Among samples with drug resistance mutations, in all cases, mutations associated with pharmacological resistance to two or three drug groups were found. Given the high incidence of resistance mutations in patients on ineffective ART, surveillance of HIV-1 drug resistance, in both ART-receiving and ART-naive individuals, appears necessary. A lack of vigilance and control measures may lead to the spread of primary ART-resistant HIV strains. [ABSTRACT FROM AUTHOR]

Osteoarthritis is the most common joint disease in animals, which is not always treatable. Both in veterinary and in human medicine, laboratory research methods are used to effectively diagnose this pathology. The purpose of this study was to establish the dynamics of markers of the acute phase of inflammation in the blood using experimental modelling of knee osteoarthritis in rabbits. Artificial reproduction of the pathological process in the knee joints of animals was performed by intra-articular administration of Yellow peel 2*5 ml. The BC-2800Vet automatic haematology analyser (China) was used to calculate the number of white blood cells and their subpopulations in the blood. The content of C-reactive protein in blood serum was found using enzyme-linked immunosorbent assay. The rate of erythrocyte sedimentation in the blood was investigated according to the Panchenkov method. Thus, on Day 7, a sharp increase in the number of white blood cells (2.7 times) and their subpopulations was noted in sick rabbits, which indicates an active inflammatory process of a systemic nature. On Day 14, their number in the blood of sick animals was characterized by a tendency to decrease, but it had not yet reached the values of the control group. On Days 21 and 28, all the indicators under study tended to decrease in sick animals. Specifically, the number of white blood cells and their subpopulations acquired reference values. The content of C-reactive protein significantly increased by 40.8 times in sick animals on Day 7 of the study and decreased by 1.7 times on Day 14 compared to its values at the previous stage. At the same time, on Day 28, this indicator increased by 19.3 times compared to the indicators of the control group of animals. The rate of erythrocyte sedimentation in the blood of sick rabbits significantly increased by 2.3 times compared to the control group. On Day 21, this indicator in the blood of sick rabbits decreased by 1.4 times compared to its values on Day 14 of the experiment. On the other hand, on Day 28, its value increased by 1.7 times compared to the control (4.3 mm/h). The results obtained will contribute to further targeted laboratory diagnostics of knee osteoarthritis in animals, which will ensure the proper effectiveness of therapeutic and preventive measures and improve their overall health [ABSTRACT FROM AUTHOR]

Rationale. Countries of Africa, especially countries in sub-Saharan Africa, represent a region characterized by high incidence of chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Methods for detection of HBV and HCV in low and middle-income countries differ from those that are used in countries having access to high-cost technologies. The Republic of Guinea is a region with high prevalence of hepatotropic viruses, however, the information on HBV and HCV prevalence in the area is extremely limited, thus emphasizing the significance of this study.The purpose of the study is to evaluate the need for improving laboratory diagnostics of parenteral HBV and HCV infections in the Republic of Guinea.Materials and methods. A total of 2,616 samples of blood serum were tested, the samples were collected from apparently healthy residents of the Republic of Guinea during the routine medical checkup. The testing included qualitative detection of HBsAg, anti-HBs IgG, anti-HBcore IgG, anti-HCV IgG antibodies as well as HBV DNA and HCV RNA.Results. The detection frequency of serological markers of HBV and HCV infections was 80.77% and 18%, respectively. However, HBsAg was detected only in 16.01% of individuals. Tests for detection of HBV DNA were performed among seropositive patients and patients seronegative by other HBV markers, HBV DNA was detected in 22.36% of cases, including 6.07% of HBsAg-negative cases. HCV RNA was detected in 2.2% of cases. Both HCV RNA and HBV DNA were detected in 27 people, including 19 HBsAg-negative cases, thus accounting for 1.03% of the examined group.Conclusions. The markers that are currently used for laboratory detection of HBV and HCV in the Republic of Guinea are not efficient enough to diagnose reliably all cases. Undoubtedly, there is an urgent need to improve laboratory diagnostics for timely detection of parenteral viral hepatitis. Routine laboratory operations need assays for additional serological and molecular markers of HCV and HBV infections.

Blood α-amylase activity is a key laboratory marker of pancreatic diseases. At the same time, some of the common laboratory methods of its measurement have a number of limitations. Therefore new more effective laboratory methods are currently being developed over those methods that have been already available. The aim of this work was to compare two methods of determination of the activity of total and pancreatic α-amylase in human blood based on using 4,6-ethylidene-(G7)-p-nitrophenyl-(G1)-α,D-maltoheptaoside (EPS-G7) and 2-chloro-4-nitrophenyl-4-O-β-D-galactopyranosylmaltoside (GalG2CNP) as substrates. Blood serum samples from patients with different levels of α-amylase activity were analyzed. The main analytical characteristics of the method with the GalG2CNP substrate were determined using purified α-amylase specimens. A high-sized correlation and high accuracy of the α-amylase isoenzymes activity were observed for the both methods. Therefore, the method for determining the activity of α-amylase using GalG2CNP as a substrate has analytical characteristics similar to the common laboratory method with EPS-G7, but at the same time, the existing advantages allow it to be recommended for wider application in clinical laboratory diagnostics and scientific research.

The review considers the relevant aspects of laboratory diagnosis of rabies. The methods of laboratory diagnostics of rabies infection, standardized by WHO in 2018, and their use in the Russian Federation are presented. The scheme of laboratory diagnostics of rabies, applied by specialists of the “48th CRI” of the Ministry of Defense of Russia, for the study of biological samples from deceased people is outlined. Between 2002 and 2018, the study of biomaterial was carried out using molecular-biological, virological methods of diagnosis and in some cases electron microscopy, which allowed to detect and identify the pathogen in 257 samples from 71 people, to certify and deposit new isolates of the rabies virus. Accumulation and analysis of the lessons learned in the application of molecular-biological method of rabies diagnosis allows us to recommend the use of RT-PCR, real-time RT-PCR sets (having a certificate of state registration) in healthcare and veterinary medicine practice to identify the causative agent of rabies infection. The use of molecular-biological methods is promising in terms of the development of rabies diagnosis to improve epidemiological surveillance and raise the efficiency of the biological protection of the population of the Russian Federation.

Aleksey Chaulin1,2 1Department of Cardiology and Cardiovascular Surgery, Samara State Medical University, Samara, 443099, Russia; 2Department of Histology and Embryology, Samara State Medical University, Samara, 443099, RussiaCorrespondence: Aleksey ChaulinDepartment of Cardiology and Cardiovascular Surgery, Samara State Medical University, 89 Chapaevskaya Street, Samara Region, Samara, 443099, RussiaTel +7 (927) 770 25 87Email alekseymichailovich22976@gmail.comAbstract: Laboratory diagnosis plays one of the key roles in the diagnosis of many diseases, including cardiovascular diseases (CVD). The methods underlying the in vitro study of many CVD biomarkers, including cardiac troponins (cTnI and cTnT), are imperfect and are continually being improved to enhance their analytical performance, with sensitivity and specificity being the most important. Recently developed improved cTnI and cTnT detection methods, referred to as highly sensitive methods (hs-cTnI, hs-cTnT), have changed many of our ideas about the biology of cardiac troponins and opened up a number of additional diagnostic capabilities for practical healthcare. This article systematizes some relevant data on the biology of cardiac troponins as well as on methods for determining cTnI and cTnT with an analysis of the diagnostic value of their analytical characteristics (limit of blank, limit of detection, 99th percentile, coefficient of variation, and others). Data on extracardiac expression of cTnI and cTnT, mechanisms of formation and potential clinical significance of gender, age, and circadian characteristics of hs-cTnI and hs-cTnT content in serum are discussed. Considerable attention is paid to the discussion of new diagnostic capabilities of hs-cTnI, hs-cTnT, including consideration of promising possibilities for their study in biological fluids that can be obtained by non-invasive methods. Also, some possibilities of using hs-cTnI and hs-cTnT as prognostic laboratory biomarkers in healthy people (for example, to assess the risk of developing CVD) and in patients suffering from a number of pathological conditions that cause damage to cardiomyocytes are examined, and the potential mechanisms underlying the increase in hs-cTnI and hs-cTnT are discussed.Keywords: laboratory diagnostics, cardiovascular diseases, acute myocardial infarction, biomarkers, cardiac troponins, cTnI, cTnT highly sensitive methods, hs-cTnI, hs-cTnT, analytical characteristics, 99th percentile, limit of detection, coefficient of variation

Melioidosis is a life-threatening infection caused by Burkholderia pseudomallei, an environmental Gram-negative bacterium, inhabitant of moist soils in the tropics and subtropics. There is no licensed vaccine against melioidosis. The main routes of B. pseudomallei infection are percutaneous inoculation, inhalation, or ingestion. Individual cases of vertical, sexual, zoonotic, and nosocomial transmission of melioidosis are described. Risk factors for infection are contact with soil or water (especially during the rainy season). The age over 45, type 2 diabetes, alcoholism, liver disease, chronic lung disease, chronic renal disease, and thalassemia, as well as long-term use of steroids and immunosuppressive therapy, are the main susceptibility factors for melioidosis. Among the affected adult residents of endemic regions, 80% had one or more predisposing factors, among children — about 20%. No significant influence of concomitant diseases on the development of melioidosis in travelers was found. Less than 50% of patients had predisposing factors. The incubation period of melioidosis ranges within 1—21 days; on average, 9 days, in case of sizeable infectious dose, it can be less than one day. There is no post-infectious immunity, and reinfection can occur with a different B. pseudomallei strain after successful treatment. B. pseudomallei is a facultative intracellular pathogen that can invade and multiply inside a wide range of cells, including phagocytic. The acute form of melioidosis is characterized by pneumonia, multiple abscesses, bacteremia, and systemic sepsis. Chronic, subacute, and latent forms are also possible. Antimicrobial therapy is divided into the initial intensive phase and the subsequent eradication phase. B. pseudomallei is resistant to penicillins, first- and second-generation cephalosporins, aminoglycosides, macrolides, chloramphenicol, fluoroquinolones, tetracyclines, trimethoprim, and in some cases to co-trimoxazole, and rarely to ceftazidime. Early diagnosis and appropriate management are crucial in reducing severe complications leading to high mortality, and in preventing disease recurrences. However, there is no pathognomonic melioidosis-specific feature, and the disease is not well known to physicians and microbiologists. The results of serological tests for detection of specific antibodies are ambiguous. The bacterial load of the tested sample limits the detection of antigens. Among the accelerated methods for identifying the causative agent of melioidosis, PCR has the highest sensitivity and specificity. Automated identification using microbiological analyzers generally shows good results, but about 15% of isolates are misidentified. Time-of-flight mass spectrometry with matrix-assisted laser desorption ionization is potentially useful for rapid identification of B. pseudomallei. However, existing databases require optimization by adding the reference spectra for B. pseudomallei.

Introduction. Colorectal cancer is one of the most common cancers worldwide. In the Republic of Kazakhstan, over the past decade, the incidence rates of colon and rectal cancers have increased by 23 % and 17 %, respectively. Although colorectal cancer screening rates have increased over time, it is necessary to identify new serum markers for early detection and prognosis of this cancer. The aim of the study was to evaluate the diagnostic value of serum biomarkers for colorectal cancer. Material and Methods. Biomarkers were studied by immunofluorescence method using xMap technology, which included simultaneous immunofluorescence determination on magnetic spheres CA125, CA15–3, CA19–9, CEA, CYFRA21–1, sFAS, sFasL, HE4, Osteopontin, VEGF-A, pd-L1. Results. The study showed no significant differences in the levels of markersbetween healthy subjects (control group) and patients with bowel diseases. In cancer patients, СEA and OpN markers increased with increasing tumor stage, and were significantly higher than those observed in patients with bowel diseases and controls. The CA125 marker was significantly higher in patients with stage IV cancer then in patients with I–III stages. Conclusion. Our study revealed that the new pd-L1 marker can serve as a significant biomarker for colorectal cancer.

In order to mitigate the spread of COVID-19, in the early stages of the pandemic outbreak, postponing elective procedures was recommended all around the world. Outpatient care and dental care were limited to telephone advice and emergency services. Dental staff is particularly vulnerable to SARS-CoV-2 contraction, because of the inevitable contact with patients’ body fluids during aerosol-generating procedures. The implementation of diagnostic tests among ambulatory patients could improve the occupational safety among outpatient care personnel. The aim of this review was to introduce information regarding COVID-19 diagnostics with a particular focus on the methods which can be utilized in an outpatient and dental care setting. An online PubMed database review of articles on COVID-19 diagnostics, published on February 12–May 15, 2020, was conducted. Reverse transcription polymerase chain reaction is the gold standard in COVID-19 diagnostics, which determines if a person has an active infection. Unfortunately, its utilization in outpatient care is limited. Serological enzyme-linked immunosorbent assays identify people who were infected, including those who have had an asymptomatic infection, but they do not give sufficient information about the acute infection. Rapid serological assays developed to facilitate testing outside of laboratories, especially in dental offices, are not recommended by the World Health Organization to be used outside research settings, and they should not constitute the basis for clinical decision-making because of frequent false-negative results which may consequently contribute to personnel infections. Out of all available COVID-19 diagnostic methods, rapid serological assays seemed to be a method of choice in outpatient medical care. Unfortunately, their results turned out to be unreliable. The best methods to ensure the occupational safety of medical staff and to avoid cross-infections in outpatient care facilities include a thorough epidemiological interview, temperature measurement to rule out patients with an active infection, and the implementation of strict infection control procedures. Med Pr. 2021;72(2):155–62

The aim of our work was to compare the HBV, HCV and HIV vertical transmission risk in the Republic of Guinea.Materials and methods. The material for the study was 305 blood plasma samples from pregnant women living in Conakry, Republic of Guinea. The samples were examined for the presence of serological (HBsAg, antibodies antiHBs IgG, anti-HBcore IgG, anti-HCV IgG, Ag/Ab-HIV) and molecular (HBV DNA, HCV RNA, HIV RNA) markers.Results and discussion. When assessing the overall prevalence of serological markers among patients, the incidence of HBV markers was 76.06 %. Antibodies to HCV were detected only in 1 case, which amounted to 0.32 %. HIV markers were detected in 3 cases, which amounted to 0.98 %. The prevalence of HBsAg in the group under examination significantly differed between the groups of pregnant women aged 13–19 years (17.33 %) and 20–24 years (12.12 %), p

Genomic data enable the development of new biomarkers in diagnostic laboratories. Examples include data from gene expression analyses or metagenomics. Artificial intelligence can help to analyze these data. However, diagnostic laboratories face various technical and regulatory challenges to harness these data. Existing software for genomic data is usually designed for research and does not meet the requirements for use as a diagnostic tool. To address these challenges, we recently proposed a conceptual architecture called "GenDAI". An initial evaluation of "GenDAI" was conducted in collaboration with a small laboratory in the form of a preliminary study. The results of this pre-study highlight the requirement for and feasibility of the approach. The pre-study also yields detailed technical and regulatory requirements, use cases from laboratory practice, and a prototype called "PlateFlow" for exploring user interface concepts. [ABSTRACT FROM AUTHOR]