Clinical Diagnostic Laboratories Research Articles

Клініко-анамнестичні характеристики та медикаментозний супровід вагітних жінок із обтяженим акушерським анамнезом і тромбофіліями

The purpose of the study was to determine the clinical and anamnestic features (age characteristics of women and anamnestic features of previous pregnancies) of pregnant women with burdened obstetric history and thrombophilia with the establishment of the main pharmacological mechanisms of therapeutic support. Materials and methods. The study was performed at the Department of Clinical Laboratory Diagnostics of Kharkiv National Medical University and at the Kharkiv Clinical Laboratory Center in the period from 2017 to 2021. We examined 137 pregnant women: the main group (101 women – pregnant women with a burdened obstetric history and thrombophilia aged 29.0 to 37.0 years) and the control group (36 women – pregnant women with a burdened obstetric history without thrombophilia aged 26.0 to 30.0 years). In general, the predominance of one (35.8 %) and two (46.7 %) pregnancies in the history of women with a burdened obstetric history compared to three (17.5 %) was noted. Results and discussion. An increase (U = 706.0; p <0.001) in the chances of developing negative consequences of pregnancy in thrombophilia with increasing age due to negative idiopathic thromboembolic effects due to thrombophilia was noted. A higher mean age of pregnant women with a burdened obstetric history in the presence of thrombophilia (33.0 [29.0; 37.0] years) was determined compared with the control (28.0 [26.0; 30.0] years). The maximum negative manifestations of idiopathic thromboembolic effects due to thrombophilia in the first pregnancy were found. The predominance of cases of fetal death in the first pregnancy of women with two (94.1 %) and three (78.3 %) pregnancies was determined. In general, the need for antiplatelet therapy in thrombophilia was stated. Its use (χ2 = 46,950; p <0.001) was determined in 85.1 % of cases, the majority (65.3 %) of one drug (probably (χ2 = 164.54; p <0.001) more often (52.5 %) acetylsalicylic acid compared with dipyridamole (5.9 %)). It was found that anti aggregate therapy, which included two drugs (combination: acetylsalicylic acid, dipyridamole or klexan) was most often used in the first (40.7%) pregnancy in contrast to the second (11.8%) and third (13.0 %). Conclusion. The study found that antiplatelet therapy with only one drug (acetylsalicylic acid, dipyridamole or klexan) was more commonly used in the second (74.5%) and third (73.9%) pregnancies than in the first one (40.7%). It was naturally determined that the absence of any medication was noted most often by pregnant women in the first (18.5%) pregnancy in contrast to the second (13.7%) and third (13.0%)

EP474: Prenatal exome sequencing analysis in the clinical setting of fetuses with structural anomalies or increased nuchal translucency

Although prenatal screening and diagnosis are generally mostly focused on the detection of aneuploidies and subchromosomal abnormalities, testing for monogenic disorders has become an important prenatal diagnostic test for pregnancies complicated by congenital anomalies. Multiple studies using gene panels and exome sequencing of fetal samples have revealed that pathogenic variants in single genes are an important cause of congenital anomalies. In this study, we reviewed the indications and results of prenatal trio-exome sequencing performed in a clinical diagnostic laboratory.

EP374: Variant classification changes over time at a clinical molecular diagnostic laboratory

As genetic testing increasingly informs clinical decision making across medical specialties, accurate variant interpretation is essential to ensure patients receive appropriate care. Current guidelines for genomic sequence variant interpretation delineate the evidence that should be incorporated in variant assessments as well as how different types of evidence should be weighted. Variants can be re-interpreted and reclassified over time as new evidence accrues. The Advanced Molecular Diagnostics Laboratory (AMDL) at Mount Sinai Hospital in Toronto, Ontario, provides publicly funded multigene panel testing for patients with suspected hereditary disorders.

OP034: Evaluation of the 2019 ACMG-ClinGen guidelines for interpretation of copy-number variants with borderline classifications at an academic clinical diagnostics laboratory

Copy-number variation is a source of both genetic diversity and Mendelian disease. Analysis of copy-number variants (CNVs) is an essential and frequently utilized component of medical genetics for a wide variety of patient indications. In 2019, the American College of Medical Genetics and Genomics (ACMG) and the Clinical Genome Resource (ClinGen) developed new technical standards for the interpretation of CNVs, introducing a semi-quantitative classification system that aims to foster greater standardization and consistency of classifications between laboratories.

Development and Validation of Two RT-qPCR Diagnostic Assays for Detecting Severe Acute Respiratory Syndrome Coronavirus 2 Genomic Targets across Two Specimen Types

Following the outbreak and subsequent pandemic of coronavirus disease 2019 (COVID-19), clinical diagnostic laboratories worldwide sought accurate and reliable testing methodologies. However, many laboratories were and still are hindered by a number of factors, including an unprecedented demand for testing, reagent and laboratory supply shortages and availability of qualified staff. To respond to these concerns, two separate laboratory-developed tests were validated for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using two different specimen types. In addition, these assays target different genomic regions of SARS-CoV-2, allowing for viral detection and mitigating genetic variation. Lower limit of detection and clinical evaluation studies showed detection of SARS-CoV-2 at 500 cp/mL with nasopharyngeal and saliva samples. These multiplexed RT-qPCR assays, although based on modified CDC, New York State Department of Health, and World Health Organization Emergency Use Authorization tests, allow for higher throughput and rapid turnaround time, benefiting patients, clinicians, and communities as a whole. These cost-effective tests also use readily obtainable reagents, circumventing commercial assay supply chain issues. The laboratory-developed tests described here have improved patient care and are highly adaptable should the need arise at other clinical diagnostic laboratories. Furthermore, the foundation and design of these assays may be modified in the future for detection of COVID-19 variants or other RNA-based viral detection tests.

Deer Pasture Productivity and Crop Yield in Anabarsky Ulus of the Republic of Sakha (Yakutia)

This article presents the results of the research into the crop yield and productivity of deer pastures in the Republic of Sakha (Yakutia). Currently, the policies aim to preserve and increase the headcount of local breeds of domesticated reindeer. Due to this, we researched the food resources of deer pastures in Anabarsky Ulus. Laboratory research and livestock analysis of the food resources of deer pastures were carried out at the clinical diagnostic research laboratory of the Arctic State Agrotechnological University. The analysis of soil was carried out at the laboratory of the Agricultural Service State Budget Company under the auspices of the Ministry of Agriculture of the Republic of Sakha (Yakutia). The authors prove that the crop yield of natural deer pastures reaches up to 41.3 dt/ha at the total power of 38.4 GJ/ha, 2891 fodder units, and 198 kg of raw protein. Day-long deer pasturing plays an important role in preserving pastures. This system stipulates that the herdsmen actively control the herd and change the pastures daily depending on the deer’s need in grazing and having rest and taking into account the weather conditions, as well as the availability and quality of fodder.

Ultrasonic disintegration of circulating immune complexes is a clinical innovation laboratory diagnostics of infectious diseases

Data describing the effects of ultrasound upon blood serum biological objects are analyzed in the paper. Biophysical as well as medical aspects of tissue reactions caused by ultrasound are considered in fine detail including the immune complexes’ dissociation processes accompanied by releasing of antibodies and antigens that used to be present in bound form and have been inaccessible for revelation using laboratory tests. The prospects of using ultrasonic techniques in laboratory diagnostics are presented using HIV infection as an example.

Serologic evidence of exposure to Human Immunodeficiency Virus Type 1 and Dengue Virus among patients attending diagnostic Laboratory, Lagos State, Nigeria

Reportedly, in co-infection, dengue viral (DENV) replication negatively impacts human immunodeficiency virus type 1 (HIV-1). Reports on this or on people having dual HIV-1/DENV are however, scarce in Nigeria, where both HIV-1 and DENV are endemic. As a preliminary investigation in southwest Nigeria, we hypothesized that a high proportion of individuals presented for laboratory tests have serologic evidence of dual HIV-1/DENV infection. This is a cross-sectional Research Institute-based serologic study; patients attending a specific Clinical Diagnostic Laboratory in Lagos State were enrolled. Plasma samples (n=150) of consenting asymptomatic patients were serially screened for evidence of HIV-1 antibodies using DETERMINETM and UNI-GOLDTM HIV-1/-2 rapid test kits. Only the samples (n=54) confirmed HIV-1 reactive by the latter (and some randomly selected non-reactive samples [n=37]) were tested with ELISA kit for anti-DENV immunoglobulin G (IgG) antibodies. Microsoft Excel and Statistical Package for the Social Sciences were used for statistical analysis. Some participants had detectable HIV-1/DENV antibodies with dual positivity rate of 16.5% [as group-specific point-prevalence rate]. This was independently associated (p=0.04) with marital status as participants who were neither single nor married (categorized as ‘others’) had more than 11 times (OR: 11.3) likelihood of being dual seropositive. Mono-positivity of HIV-1 antibody among the 91 participants tested for dual HIV-1/DENV seropositivity was 59.3%; corresponding rate for DENV antibody was 20.9%. As per mono-positivity rates, the likelihood of participants testing positive to HIV-1 antibody was more than 21 times (Odds ratio [OR]: 21.3) that of DENV antibody. In conclusion, 15 participants (16.5%) of this specific group had serologic evidence of dual HIV-1/DENV infection; this we considered low given the endemicity of Nigeria for both viruses. As a subpopulation with unusually high HIV-1 mono-positivity; thelow DENV mono-positivity might be responsible for the low dual HIV-1/DENV positivity.

Assessment of biosafety implementation in clinical diagnostic laboratories in Pakistan during the COVID-19 pandemic

Laboratory diagnostic capacity is crucial for an optimal national response to a public health emergency such as the COVID-19 pandemic. Preventing laboratory-acquired infections and the loss of critical human resources, especially during a public health emergency, requires laboratories to have a good biorisk management system in place. In this study, we aimed to evaluate laboratory biosafety and biosecurity in Pakistan during the COVID-19 pandemic. In this cross-sectional study, a self-rated anonymous questionnaire was distributed to laboratory professionals (LPs) working in clinical diagnostic laboratories, including laboratories performing polymerase chain reaction (PCR)-based COVID-19 diagnostic testing in Punjab, Sindh, Khyber Pakhtunkhwa, and Gilgit-Baltistan provinces as well as Islamabad during March 2020 to April 2020. The questionnaire assessed knowledge and perceptions of LPs, resource availability, and commitment by top management in these laboratories. In total, 58.6% of LPs performing COVID-19 testing reported that their laboratory did not conduct a biorisk assessment before starting COVID-19 testing in their facility. Only 31% of LPs were aware that COVID-19 testing could be performed at a biosafety level 2 laboratory, as per the World Health Organization interim biosafety guidelines. A sufficiently high percentage of LPs did not feel confident in their ability to handle COVID-19 samples (32.8%), spills (43.1%), or other accidents (32.8%). These findings demonstrate the need for effective biosafety program implementation, proper training, and establishing competency assessment methods. These findings also suggested that identifying and addressing gaps in existing biorisk management systems through sustainable interventions and preparing LPs for surge capacity is crucial to better address public health emergencies.

Inpatient Cost of Stroke Care in Greece: Preliminary Results of the Web-Based "SUN4P" Registry.

Direct medical costs (diagnostic imaging and clinical laboratory exams, overheads/bed cost, pharmaceuticals, ringers and other non-durables and inpatient rehabilitation) per patient were calculated from the providers' (hospitals') perspective. Resource use data derived from the "SUN4P" web-based registry and unit costs were retrieved from publically available sources and were assigned to resource use. The sample comprised 6,282 inpatient days of 750 patients (mean age: 75.5±13.3 years) admitted from July 2019 to July 2021, in nine public hospitals. Mean length of stay was 8.4±7.6 days and mean total operational cost was calculated to €1,239.4 (from which 45% and 35% related to diagnostic exams and overheads/bed cost respectively). Mean cost related to hemorrhagic stroke patients that were discharged alive was calculated significantly higher compared to mean cost related to ischemic stroke patients who didn't undertake thrombolysis and were also discharged alive from the hospital (€2,155.2 vs. €945.2, p<0.001). Linear regression analysis revealed that length of stay was significantly correlated with cost (coefficient beta=232, 95% CI confidence interval = 220-243, p<0.001). These findings are in accordance with current evidence and should be thoroughly assessed to rationalize inpatient reimbursement rates in order to achieve improved value of care.

Interprofessional Team Diagnostics Consultation Improves Health Professions’ Practice Outcomes and Clinical Research

Background: Communication gaps in health services delivery significantly compromise quality in clinical decision making. Information generated by diagnostics professionals’ accounts for much of the objective data in the clinical record and therefore is foundational in clinical decision support. This work describes the Diagnostics Consultation Model©, a diagnostics communications portal, which supports communications among interprofessional teams, providers, and institutions. Aims: Study aims were to develop and validate a workflow prediction index (the complexity index) to assign resolution of consultation requests to diagnostics practitioners with requisite competencies based on an algorithm comprised of characteristics available at the point of consultation initiation. The complexity index functions as the entry into a workflow process directing consultation requests, first, to diagnostics practitioners for investigation and then into communication processes for tracking medical history, patient/consumer clinical information, resolution logic, conclusions, and next step recommendations among all healthcare providers. Methods: Data to develop the complexity index (N = 325 consultation cases) were collected during daily activities in the clinical diagnostics laboratory and describe consultation characteristics important in clinical decision making and available at the point of consultation initiation. The complexity index was developed and validated by comparison of regression analyses using consultation characteristics, i.e., clinical outcomes, available at the point of consultation initiation (development) and after consultation completion (validation). Results: Diagnostics Consultation Model© methodology links communication processes among all providers in all care settings, i.e., community, institutional, and referral, involved in the care paths of individual patient/consumers. This methodology also provides the capability to follow individuals’ medical histories longitudinally and, through regular consultations and practice-based clinical research, to address issues of medical effectiveness, cost efficiency, access, equity, timeliness, safety, and compliance. Conclusion: Implementation of Diagnostics Consultation Model© methods and curriculum in health professions’ daily practice has the potential to change health services delivery by the redistribution of care through interprofessional teams coordinated by standardized communication processes. Employed as a systems approach to individualized patient/consumer care, the Diagnostics Consultation Model© could provide the communications technology and methodology structure for value-based healthcare continuously optimized to address the needs of individuals, populations, and health systems throughout the continuum of care.

Верификация цефтриаксона и определение его концентрации в сыворотке крови методом капиллярного электрофореза у больных хирургического профиля

Objective: to develop a method for verifying ceftriaxone and determining its concentration in blood serum by means of capillary electrophoresis. Materials and methods. The study used ceftriaxone sodium salt (CJSC Pharmaceutical Company "Lekko"), model mixtures of biological fluids (whole blood, serum and blood plasma) with different concentrations of ceftriaxone; clinical biological blood samples taken during breast surgery against the background of perioperative antibiotic prophylaxis. The concentration of ceftriaxone was determined by using a Capel-105M capillary electrophoresis system (CJSC Lumex, St. Petersburg, Russia). Results. An express method has been developed for the determination of ceftriaxone in blood serum (sample volume - 0.1 ml), which involves the use of a precipitant of protein substances - crystalline ammonium sulfate, extraction of the organic system chloroform - isobutanol in a ratio of 3: 1, re-extraction of the substance into a working electrolyte diluted with water, and subsequent electrophoresis on the device "Capel-105M". Method validation indicates its linearity, correctness, precision and specificity; the lower limit of quantitation is 0.1 µg/0.1 ml biological medium. The effectiveness of the technique was confirmed in a clinical trial by determining the concentration of ceftriaxone in the blood serum 30 minutes after intravenous administration of 1 g of the drug for the purpose of perioperative antibiotic prophylaxis. Conclusion. The method for the quantitative determination of ceftriaxone in blood serum, performed by using a capillary electrophoresis system (“Kapel-105M”), meets the established validation criteria. At the same time, the identification of the antibiotic is carried out under selective conditions and requires a minimum amount of the object, thereby reducing the amount of co-extractive substances in the samples, which, in combination with the method of stacking during electrophoresis, provides a high sensitivity of the technique. It can be recommended for analysis in clinical diagnostic laboratories.

A CASE OF ANEMIA OF MIXED GENESIS WITH A DISCREPANCY IN THE RESULTS BETWEEN THE 3-DIFF HEMATOLOGICAL ANALYZER AND LIGHT MICROSCOPY

A case of severe anemia of mixed genesis is presented. The clinical picture of the disease is described, the data of laboratory investigations are given. The interest of this case for attending physicians and employees of clinical diagnostic laboratories (CDL) is due to significant discrepancies between the results of blood analysis according to automatic analysis performed using a hematology analyzer and the data of light microscopy of a stained blood smear. Various morphological changes were found in the stained blood smear, which contradicted relatively normal erythrocyte parameters (MСV, MСН, RDW) and the absence of any flags according to the hematological analyzer. The problems of using hematological analyzer data as a basis for differential diagnosis of anemia with various disorders of erythrocyte and leukocyte morphology, especially when using artificial intelligence for decision-making, are discussed.

RNAscope for VEGF-A Detection in Human Tumor Bioptic Specimens.

Different pro-angiogenic factors, such as vascular endothelial growth factor-A (VEGF-A), have been related to microvascular density, clinicopathologic factors, and poor prognosis in many tumors. VEGF-A binds its receptor2 (VEGFR2) to induce neo-angiogenesis, a constant hallmark of tumor initiation and progression. Based on VEGF-A/VEGFR2 relevance in tumor angiogenesis, several inhibitors were developed. However, the clinical benefits of anti-angiogenic therapies are limited because tumors activate different mechanisms of drug resistance.The need for understanding tumor biology, limitation or failure of anti-angiogenic therapies, and the demand for a personalized therapeutic approach has boosted the search for robust biomarkers for patient stratification as responder or non-responder to anti-VEGF therapies.This chapter presents a detailed protocol to perform chromogenic VEGF-A mRNA detection and quantification in human tumor bioptic specimens using RNAscope technology and RNA-in situ hybridization (ISH) algorithm. RNAscope for VEGF-A detection, even for small amounts, is compatible with precious clinical samples and diagnostic laboratory workflows.

Immunoglobulin/T Cell Receptor Capture Strategy for Comprehensive Immunogenetics.

In the era of genomic medicine, targeted next generation sequencing strategies (NGS)are becoming increasingly adopted by clinical molecular diagnostic laboratories to identify genetic diagnostic and prognostic biomarkers in hemato-oncology. We describe the EuroClonality-NGS DNA Capture (EuroClonality-NDC) assay, which is designed to simultaneously detect B and T cell clonal rearrangements, translocations, copy number alterations, and sequence variants. The accompanying validated bioinformatics pipeline enables production of an integrated report. The combination of the laboratory protocol and bioinformatics pipeline in the EuroClonality-NDC minimizes the potential for human error, reduces economic costs compared to current molecular testing strategies, and should improve diagnostic outcomes.

A Rare Case of Viral Dermatitis Induced Reactive Histiocytosis in a Dog

A male Labrador aged 2 years was presented to Multi-speciality Veterinary Hospital, Guru Angad Dev Veterinary and Animal Science University, Ludhiana. The animal was presented with a complaint of recurrent nodular skin lesions. The blood and serum samples were sent to clinical diagnostic laboratory for routine haematology and serum biochemistry to assess the health status of the dog. Impression smears, FNAC along with biopsy samples were taken from the skin lesions. Biopsy sample was processed for accomplishment of accurate result. The impression smears revealed plenty of RBC along with neutrophils. The FNAC sample revealed plenty of pleomorphic histiocytes and cells of lymphoid series suggesting histiocytosis. The histopathological slides of biopsy sample revealed presence of intranuclear inclusion bodies in the squamous cells of skin and infiltration of histiocytes. For confirmation of intranuclear inclusions, Triple shorr’s stain was done and magenta-coloured inclusion bodies were observed in the squamous cells which were suggestive of viral aetiology induced proliferation of histiocytes. Based on cytological and histopathological examination, it was concluded that the dog suffered from virus induced skin lesions which led to proliferation of histiocytes and hence, cutaneous reactive histiocytosis.

Assessment of the readiness of medical personnel of the clinical diagnostic laboratory for innovative changes

Assessment of the readiness of medical personnel of the clinical diagnostic laboratory for innovative changes

Use of the WISN method to assess the health workforce requirements for the high-volume clinical biochemical laboratories

BackgroundThe clinical laboratory services, as an essential part of health care, require appropriate staff capacity to assure satisfaction and improve outcomes for both patients and clinical staff. This study aimed to apply the Workload Indicators of Staffing Need (WISN) method for estimating required laboratory staff requirements for the high-volume clinical biochemical laboratories.MethodsIn 2019, we applied the WISN method in all 13 laboratories within the Center for Medical Biochemistry of the University Clinical Centre of Serbia (CMB UCCS). A review of annual routinely collected statistics, laboratory processes observations, and structured interviews with lab staff helped identify their health service and additional activities and duration of these activities. The study outcomes were WISN-based staff requirements, WISN ratio and difference, and a recommendation on the new staffing standards for two priority laboratory workers (medical biochemists and medical laboratory technicians).ResultsMedical biochemists’ and laboratory technicians’ annual available working time in 2019 was 1508 and 1347 working hours, respectively, for the workload of 1,848,889 samples. In general, the staff has four health service, eight support, and 15 additional individual activities. Health service activities per sample can take from 1.2 to 12.6 min. Medical biochemists and medical laboratory technicians spend almost 70% and more than 80% of their available working time, undertaking health service activities. The WISN method revealed laboratory workforce shortages in the CMB (i.e. current 40 medical biochemists and 180 medical laboratory technicians as opposed to required 48 medical biochemists and 206 medical laboratory technicians). Workforce maldistribution regarding the laboratory workload contributes to a moderate–high workload pressure of medical biochemists in five and medical laboratory technicians in nine organizational units.ConclusionsThe WISN method showed mainly a laboratory workforce shortages and workload pressure in the CMB UCCS. WISN is a simple, easy-to-use method that can help decision-makers and policymakers prioritize the recruitment and equitable allocation of laboratory workers, optimize their utilization, and develop normative guidelines in the field of clinical laboratory diagnostics. WISN estimates require periodic reviews.

Разработка и валидация методики количественного определения фосфатидилэтанола в цельной крови

Introduction. Phosphatidylethanol (PEth) is a phospholipid slowly forming in cell membranes as a result of the phospholipase D enzymatic reaction between phosphatidylcholine and ethanol. As a result of the analysis of published scientific papers, high performance liquid chromatography with mass spectrometric detection is used to determine phosphatidylethanol as a method with high specificity, accuracy, precision and the ability to determine substances at minimum concentrations, which is fundamental for clinical laboratory diagnostics when monitoring alcohol abuse in the population. Aim. The aim of the study was to develop and validate a method for the quantitative determination of a direct marker of alcohol abuse, phosphatidylethanol, in human blood. Materials and methods. Phospholipid was quantified by high performance liquid chromatography with tandem mass spectrometry. Results of the study and their discussion. The developed method was validated for the following parameters: selectivity, lower limit of quantitation, linearity, sample transfer, accuracy, precision, matrix effect, stability. The analytical range was 0.0025 µmol/L – 3.0 µmol/L; LPCO level PEth 16:0/18:1 0.0025 was equal to µmol/l. Conclusions. The developed method for the quantitative determination of phosphatidylethanol is suitable for use in laboratory diagnostics in order to monitor the population in the analysis of alcohol abuse.

Urinalysis: Extract the Relevant Information Before Throwing it into the Drain

From ancient time urine has been considered as a substance of importance and examination for physical wellbeing. Evidences from the ancient civilizations including the Egyptian, Sumerian, Babylonian and Eastern Civilizations such as Vedic cultures support the use of urine as an index of physical and mental well-being. Classified as coloured, black, frothy, cloudy and sweet, urine used to be correlated with different disease conditions such as jaundice, kidney diseases, diabetes etc. These practices have been carried on even by the alchemists and have now formed as an integral constituent of clinical laboratory diagnostics. Modern approach to urinalysis can be credited to Dr. Richard Bright, MD, who in 1827 by performing urine examinations related to vol-ume, colour, pH, protein (but not cast) corre-lated his findings to several diseases and clini-cal picture including edema, proteinuria etc. Urinalysis combines the expertise from vari-ous disciplines including biochemistry, pa-thology, microbiology, cytology etc. Urinalysis may be used for screening, diagnosis, monitoring and prognosis due to the ease of collection in any settings. Modern day tech-niques such as molecular biology, immunolo-gy, and mass spectrometry with high resolu-tion microscopy have taken up urinalysis to explore the genetic predisposition to inherited diseases and tumor studies besides the routine diagnostics. So, variation occurs in urinalysis from the simple routine analysis, microscopic examination to highly sophisticated and ad-vanced automated analytical techniques. Uri-nalysis therefore, forms a key component of personalized medicine also integrating with proteomics, genomics, metabolomics approaches.