Clinical Diagnostic Laboratories Research Articles

Variant classification changes over time in the clinical molecular diagnostic laboratory setting

BackgroundVariant classification in the setting of germline genetic testing is necessary for patients and their families to receive proper care. Variants are classified as pathogenic (P), likely pathogenic (LP), uncertain...

Multi-center evaluation of the Research Use Only NeuMoDx monkeypox virus (MPXV) fully automated real-time PCR assay.

In this manuscript, we provide detailed in silico analysis and clinical evaluation of the assay using a large cohort of clinical samples across three academic centers in Europe and the United States. Because the assay differentiates MPXV clades I and II, this manuscript is timely due to the current need to rule out the regulated clade I by diagnostic clinical laboratories. In December 2023, and due to first report of cases of sexually transmitted clade I infections in the Democratic Republic of the Congo, when generic assays that do not differentiate the clades are used, samples are considered regulated. The assay meets the need of full automation and has a marked positive impact on the laboratory workflow.

Multicenter benchmarking of short and long read wet lab protocols for clinical viral metagenomics

Metagenomics is gradually being implemented for diagnosing infectious diseases. However, in-depth protocol comparisons for viral detection have been limited to individual sets of experimental workflows and laboratories. In this study, we present a benchmark of metagenomics protocols used in clinical diagnostic laboratories initiated by the European Society for Clinical Virology (ESCV) Network on NGS (ENNGS).A mock viral reference panel was designed to mimic low biomass clinical specimens. The panel was used to assess the performance of twelve metagenomic wet lab protocols currently in use in the diagnostic laboratories of participating ENNGS member institutions. Both Illumina and Nanopore, shotgun and targeted capture probe protocols were included. Performance metrics sensitivity, specificity, and quantitative potential were assessed using a central bioinformatics pipeline.Overall, viral pathogens with loads down to 104 copies/ml (corresponding to CT values of 31 in our PCR assays) were detected by all the evaluated metagenomic wet lab protocols. In contrast, lower abundant mixed viruses of CT values of 35 and higher were detected only by a minority of the protocols. Considering the reference panel as the gold standard, optimal thresholds to define a positive result were determined per protocol, based on the horizontal genome coverage. Implementing these thresholds, sensitivity and specificity of the protocols ranged from 67 to 100% and 87 to 100%, respectively.A variety of metagenomic protocols are currently in use in clinical diagnostic laboratories. Detection of low abundant viral pathogens and mixed infections remains a challenge, implying the need for standardization of metagenomic analysis for use in clinical settings.

Validation of an enzyme-linked immunosorbent assay test-system for bevacizumab concentration determing in biological fluids

Introduction. One of the reasons for the development of many diseases primarily malignant is the increased expression of Vascular Endothelial Growth Factor (VEGF). Bevacizumab is a drug that neutralizes the biological activity of VEGF. The molecular structure of bevacizumab is a recombinant humanized antibody. Its use reduces vascularization in the foci of increased VEGF expression which slows down tumor growth and also helps restore vision in a number of ophthalmic diseases. To determine the concentration of bevacizumab in human biological fluids a test system based on Enzyme-linked Immunosorbent Assay (ELISA) is presented.Aim. Aim of this study is the validation of this test system.Materials and methods. Blank sera of volunteers, bevacizumab solution, enzyme-linked immunosorbent assay, solid-phase sandwich ELISA kit, microplate photometer.Results and discussion. The following characteristics of the test system were determined: the lower limit of quantification is 2.0 mcg/ml, the upper calibration range is up to 200 mcg/ml, the accuracy and precision within one series and between series does not exceed 20 %, and the total error of the method – 30 %, short-term stability for samples at room temperature – 6 hours, long-term stability – 14 days at –20 °C, the ability to freeze/thaw of samples is up to three times, the ability to determine samples with a concentration above the upper calibrator after diluting in 2 times.Conclusion. The results obtained fully comply with international acceptance criteria and allow the use of the ELISA test system manufactured by LLC "Probiotek" for use in the field of clinical laboratory diagnostics.

Serotonin blood level study, anxiety-depressive states indicators and patients life quality after laparoscopic treatment of gastroesophageal reflux disease

Nowadays effective treatment of gastroesophageal reflux disease (GERD) remains a difficult and still unsolved problem. The use of various antisecretory drugs in combination with surgical treatment does not allow to achieve a long lasting improvement of the quality of life and reliable prevention of complications. In this regard, special attention is paid to study humoral factors influencing the pathogenesis of GERD, data on which will improve the results of therapeutic and surgical treatment. It is promising to study the dynamics of serotonin after laparoscopic treatment of various types of GERD, especially in combination with dyspepsia, anxiety or depressive disorders, as well as the possibility of using drugs that affect the metabolism of this hormone to correct the above mentioned disorders and achieve long lasting recovery after laparoscopic fundoplication. The aim of the work was to study the dynamics of serotonin in the blood during laparoscopic total fundoplication and analyze the correlation of this hormone with the psychological well-being of patients before and after surgical treatment of GERD. Materials and methods. Laparoscopic fundoplication by Short Floppy Nissen modification was performed in 35 patients with GERD. There were 26 (74.3 %) women, 9 (25.7 %) men. Age – 55.3 ± 11.3. The comparison group consisted of 20 healthy volunteers (women – 14 (70.0 %); men – 6 (30.0 %), average age – 56.7 ± 10.6), who did not undergo surgery. The groups were comparable by gender and age. The studies were carried out in venous blood plasma, samples of which were maintained using standard methods and stored in a low-temperature freezer at a temperature of -80 °C. Tests for serotonin (Serotonin ELISA, RE59121, IBL) were studied on the immunoenzyme complex ImmunoChem-2100 (USA) of the Department of Clinical Laboratory Diagnostics in the Zaporizhzhia State Medical and Pharmaceutical University. The concentration of the last indicators was expressed in ng/ml. The presence of concomitant anxiety, depression and level of psychological well-being was assessed using the GAD-7, PHQ-9 and SF-36 questionnaires. Statistical assessment of the research results was carried out using the Statistica for Windows 13 software package (StatSoft Inc., No. JPZ804I382130ARCN10-J). Differences were considered statistically significant at p < 0.05. Results. The serotonin level in the blood of practically healthy people was 5.8 ± 1.5 ng/ml. In the main group serotonin values before surgical treatment were 3.1 ± 1.3 ng/ml. After surgical treatment the serotonin level increased to 5.4 ± 1.8 ng/ml and was not statistically different from the level of practically healthy patients. The dynamics of serotonin levels correspond to the dynamics of the patients number with signs of anxiety and depression before and after surgery in the main group, as well as the improvement in the psychological state of patients after surgery compared to preoperative data on the scales of the “psychological component” of SF-36 health questionnaire: Vitality (VT), Social Functioning (SF), Role-Emotional (RE), Mental Health (MH). Conclusions. Laparoscopic crurorrhaphy and fundoplication in the Short Floppy Nissen modification through the mechanism of serotonin helps to improve the psychological state of patients with GERD in the postoperative period, eliminate anxiety and depression, improve quality of life and achieve a lasting antireflux effect. The obtained dynamics of serotonin levels confirms its participation in the formation of behavioral reactions in GERD, helping to improve the results of surgical treatment.

Analysis of thicknesses of blood collection needle by scanning electron microscopy reveals wide heterogeneity.

The preanalytical phase in clinical laboratory diagnostics is currently receiving more and more attention. This term describes one part of actions and aspects of the "brain-to-brain cycle" of the medical laboratory diagnostic procedure that take place before the analytical phase. However, the preanalytical activities, the handling of unsuitable samples and the reporting procedures are neither fully standardized nor harmonized worldwide. The influence of the properties of the blood collection needle must be acknowledged. In this work, we focused on the investigation of the internal structure and size of standardized 21G blood collection needles. All parameters were measured with a scanning electron microscope using a Jeol model JSM-6000PLUS. Our. The obtained data shows that the internal surfaces of the needles vary greatly from manufacturer to manufacturer (by around 35 %), and this may play an important role in influencing blood flow and even the risk of blood cell injury (especially hemolysis) during blood drawing. The differential actual needle diameters can vary greatly between needle manufactures and this variety may have a significant impact on laboratory values and may also lead to specimen rejection.

Identification of mononucleosis by automatic and microscopic methods (clinical case)

Infectious mononucleosis is an acute anthropogenic viral disease caused by the Epstein-Barr Virus (EBV). According to statistics from the Public Health Centre of the Ministry of Health of Ukraine, the number of cases in the period from 2018 to 2021 was 5,482; 5,467; 2,431; and 1,023; respectively. 5,467 people developed infectious mononucleosis in Ukraine in 2019. In particular, 863 cases were recorded in non-city dwellers, and 4,597 in children under 17. This indicates an increase in morbidity among urban residents, which determines the relevance of the study of infectious mononucleosis. The development of this disease is accompanied by fever, tonsillar pharyngitis, maculopapular skin rash and the appearance of atypical lymphocytes in the peripheral blood, which make up more than 10% of the total number of lymphocytes. Splenic rupture is the most dangerous complication of this disease. The article analyzes the clinical case of a child who complained of increased body temperature and enlarged lymph nodes. Attention is paid to the indicators of clinical blood analysis, namely to atypical mononuclear cells, which are an indicator of infectious mononucleosis. The results of a quantitative automated hematological analyzer with a microscopic (manual) method of diagnosis were analyzed. It has been established that if, when receiving the results on the analyzer, the indicators are outside the normal range, then it is necessary to use the microscopic method of counting, which will not only clarify but also expand the limits of diagnosis in making the final diagnosis. Thus, the increase in morbidity, difficulties in diagnosis and possibility of complications (splenomegaly and hepatomegaly) make it important to study infectious mononucleosis caused by EBV. This disease is an important scientific and practical problem of modern clinical laboratory diagnostics. Keywords: infectious mononucleosis, Epstein-Barr virus, atypical lymphocytes.

Transforming diagnostics: The implementation of digital pathology in clinical laboratories.

Digital pathology (DP) has emerged as a cutting-edge technology that promises to revolutionise diagnostics in clinical laboratories. This perspective article explores the implementation planning and considerations of DP in a single multicentre institution in Canada, the University Health Network, discussing benefits, challenges, potential implications and considerations for future adopters. We examine the transition from traditional microscopy to digital slide scanning and its impact on pathology practice, patient care and medical research. Furthermore, we address the regulatory, infrastructure and change management considerations for successful integration into clinical laboratories. By highlighting the advantages and addressing concerns, we aim to shed light on the transformative potential of DP and its role in shaping the future of diagnostics.

Abstract 6615: Reduced penetrance BRCA1 and BRCA2 pathogenic variants in clinical germline genetic testing

Abstract Despite the current binary clinical classification of variants (pathogenic versus benign), it is widely recognized that risk is a continuum. Prior studies have suggested the existence of reduced penetrance pathogenic variants (RPPVs), which pose challenges for identification, reporting, patient counseling and care. Through the current study, we sought to establish RPPVs as a new category of variants, with efforts focused specifically on BRCA1 and BRCA2 (BRCA). We compiled, compared, and assessed data for BRCA RPPVs from two large clinical diagnostic laboratories to establish an initial set of reduced penetrance pathogenic variants. Specifically, candidate BRCA RPPVs provided by clinical diagnostic laboratories were compiled to identify those with the highest prior likelihood of being a RPPV, based on concordant interpretations. Evidence of reduced penetrance included: 1) the identification of biallelic Fanconi Anemia-affected carriers; 2) splicing, missense, and nonsense-mediated RNA decay (NMD)-escaping variants; 3) laboratory-validated cancer history weighting models; 4) published variant-specific risk estimation data; 5) extrapolation of a reduced-risk interpretation onto close match variants that are expected to have the same effect; and 6) functional data. Sixteen candidate BRCA RPPV present in both lists from both laboratories were systematically assessed. For BRCA1, variants included c.5096G>A p.(Arg1699Gln) and variants impacting the canonical c.671 splice acceptor site. For BRCA2, variants included three frameshift (c.658_658delGT, c.9672dupA, c.9699_9702delTATG), two spliceogenic (c.8488-1G>A and c.8488-1G>T), and three missense (c.7878G>C, T, p.(Trp2626Cys); and c.9302T>G p.(Leu3101Arg)) variants. Our study is the first to clearly evaluate and establish BRCA RPPVs as a new class of variants, based on consistency in results across 16 variants using different interpretation strategies between laboratories. These variants impart a moderately increased risk of breast cancer, often considered to be between 2-4 fold, which should be considered when determining risk-informed cancer prevention strategies. Furthermore, our study establishes a framework to harmonize interpretation and standardize reporting of BRCA RPPV, by providing evidence to support this new classification for 16 variants. Further work to define clinically meaningful risk thresholds and categories for reporting BRCA RPPV are needed to personalize cancer risks in conjunction with other risk factors. Citation Format: Tuya Pal, Erin Mundt, Marcy Richardson, Elizabeth Chao, Tina Pesaran, Thomas P. Slavin, Fergus Couch, Alvaro N. Monteiro. Reduced penetrance BRCA1 and BRCA2 pathogenic variants in clinical germline genetic testing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6615.

Acetic Acid Enables Molecular Enumeration of Mycobacterium tuberculosis from Sputum and Eliminates the Need for a Biosafety Level 3 Laboratory.

Improved monitoring of Mycobacterium tuberculosis response to treatment is urgently required. We previously developed the molecular bacterial load assay (MBLA), but it is challenging to integrate into the clinical diagnostic laboratory due to a labor-intensive protocol required at biosafety level 3 (BSL-3). A modified assay was needed. The rapid enumeration and diagnostic for tuberculosis (READ-TB) assay was developed. Acetic acid was tested and compared to 4 M guanidine thiocyanate to be simultaneously bactericidal and preserve mycobacterial RNA. The extraction was based on silica column technology and incorporated low-cost reagents: 3 M sodium acetate and ethanol for the RNA extraction to replace phenol-chloroform. READ-TB was fully validated and compared directly to the MBLA using sputa collected from individuals with tuberculosis. Acetic acid was bactericidal to M. tuberculosis with no significant loss in 16S rRNA or an unprotected mRNA fragment when sputum was stored in acetic acid at 25°C for 2 weeks or -20°C for 1 year. This novel use of acetic acid allows processing of sputum for READ-TB at biosafety level 2 (BSL-2) on sample receipt. READ-TB is semiautomated and rapid. READ-TB correlated with the MBLA when 85 human sputum samples were directly compared (R2 = 0.74). READ-TB is an improved version of the MBLA and is available to be adopted by clinical microbiology laboratories as a tool for tuberculosis treatment monitoring. READ-TB will have a particular impact in low- and middle-income countries (LMICs) for laboratories with no BSL-3 laboratory and for clinical trials testing new combinations of anti-tuberculosis drugs.

Investigating Neonatal Multisystem Inflammatory Syndrome Associated with COVID-19 in the Neonatal Intensive Care Unit: A Case Series

Background: The COVID-19 pandemic, caused by the novel SARS-CoV-2 virus, has presented significant challenges to global healthcare systems, affecting individuals of all ages, including neonates. Recent studies have identified a condition similar to Multisystem Inflammatory Syndrome in Children (MIS-C) occurring in neonates, termed MIS-N (Multisystem Inflammatory Syndrome in Neonates), linked to maternal infection with SARS-CoV-2. Understanding MIS-N's clinical manifestations, laboratory findings, and treatment outcomes is crucial for improving neonatal care during the ongoing pandemic. Objective: The objective of this study was to explore the clinical characteristics, laboratory diagnostics, and treatment outcomes of neonates diagnosed with MIS-N, aiming to contribute to better diagnostic and management approaches for this vulnerable population. Methods: This retrospective case series analyzed neonates admitted with suspected MIS-N to the Children's Medical Center in Tehran, Iran, from March to September 2020. Inclusion criteria encompassed neonates showing clinical signs suggestive of MIS-N, elevated inflammatory markers, and positive for SARS-CoV-2 antibodies. Key data points included demographic information, clinical presentation, laboratory results (including complete blood count, inflammatory markers, and specific COVID-19 serology), treatment modalities, and patient outcomes. The effectiveness of interventions was evaluated based on symptom resolution and normalization of lab values. Results: Two neonates were identified with MIS-N. Case I involved a 39-week-old male presenting with diarrhea, dehydration, fever, and a rash, with initial inflammatory markers showing a white blood count (WBC) of 23,000 cells/μL, C-reactive protein (CRP) of 93 mg/L, and serum IgG of 10 g/L. Following treatment with corticosteroids, the patient's condition stabilized, with a decrease in WBC to 18,000 cells/μL and CRP to 34 mg/L. Case II, a 38-week-old female, exhibited a cough, rashes, with WBC of 17,000 cells/μL, CRP of 2 mg/L, and serum IgG of 40 g/L. A single dose of hydrocortisone led to symptom resolution. Both cases had no significant adverse outcomes at a three-month follow-up. Conclusion: Our findings illuminate the clinical and laboratory characteristics of MIS-N in neonates, emphasizing the role of maternal SARS-CoV-2 infection in its etiology. The successful resolution of symptoms with corticosteroid treatment highlights the potential efficacy of this intervention in managing MIS-N. These insights contribute to the growing understanding of MIS-N and underscore the need for ongoing research to define optimal diagnostic criteria and treatment protocols.

The value of preoperative diagnostic testing and geriatric assessment in frail institutionalized elderly with a hip fracture; a secondary analysis of the FRAIL-HIP study.

The aim of this study was to provide a comprehensive overview of (preoperative and geriatric) diagnostic testing, abnormal diagnostic tests and their subsequent interventions, and clinical relevance in frail older adults with a hip fracture. Data on clinical consultations, radiological, laboratory, and microbiological diagnostics were extracted from the medical files of all patients included in the FRAIL-HIP study (inclusion criteria: hip fracture, > 70years, living in a nursing home with malnourishment/cachexia and/or impaired mobility and/or severe co-morbidity). Data were evaluated until hospital discharge in nonoperatively treated patients and until surgery in operatively treated patients. A total of 172 patients (88 nonoperative and 84 operative) were included, of whom 156 (91%) underwent laboratory diagnostics, 126 (73%) chest X-rays, and 23 (13%) CT-scans. In 153/156 (98%) patients at least one abnormal result was found in laboratory diagnostics. In 82/153 (50%) patients this did not result in any additional diagnostics or (pharmacological) intervention. Abnormal test results were mentioned as one of the deciding arguments for operative delay (> 24h) for 10/84 (12%) patients and as a factor in the decision between nonoperative and operative treatment in 7/172 (4%) patients. A large number and variety of diagnostics were performed in this patient population. Abnormal test results in laboratory diagnostics were found for almost all patients and, in majority, appear to have no direct clinical consequences. To prevent unnecessary diagnostics, prospective research is required to evaluate the clinical consequences and added value of the separate elements of preoperative diagnostic testing and geriatric assessment in frail hip fracture patients.

Identification of closely related species in Aspergillus through Analysis of Whole-Genome.

The challenge of discriminating closely related species persists, notably within clinical diagnostic laboratories for invasive aspergillosis (IA)-related species and food contamination microorganisms with toxin-producing potential. We employed Analysis of the whole-GEnome (AGE) to address the challenges of closely related species within the genus Aspergillus and developed a rapid detection method. First, reliable whole genome data for 77 Aspergillus species were downloaded from the database, and through bioinformatic analysis, specific targets for each species were identified. Subsequently, sequencing was employed to validate these specific targets. Additionally, we developed an on-site detection method targeting a specific target using a genome editing system. Our results indicate that AGE has successfully achieved reliable identification of all IA-related species (Aspergillus fumigatus, Aspergillus niger, Aspergillus nidulans, Aspergillus flavus, and Aspergillus terreus) and three well-known species (A. flavus, Aspergillus parasiticus, and Aspergillus oryzae) within the Aspergillus section. Flavi and AGE have provided species-level-specific targets for 77 species within the genus Aspergillus. Based on these reference targets, the sequencing results targeting specific targets substantiate the efficacy of distinguishing the focal species from its closely related species. Notably, the amalgamation of room-temperature amplification and genome editing techniques demonstrates the capacity for rapid and accurate identification of genomic DNA samples at a concentration as low as 0.1 ng/μl within a concise 30-min timeframe. Importantly, this methodology circumvents the reliance on large specialized instrumentation by presenting a singular tube operational modality and allowing for visualized result assessment. These advancements aptly meet the exigencies of on-site detection requirements for the specified species, facilitating prompt diagnosis and food quality monitoring. Moreover, as an identification method based on species-specific genomic sequences, AGE shows promising potential as an effective tool for epidemiological research and species classification.

Detection rates of molecular alterations in vascular malformations in a clinical diagnostic laboratory

Detection rates of molecular alterations in vascular malformations in a clinical diagnostic laboratory

FDA's proposed rule for the regulation of laboratory-developed tests.

In October 2023, the Food and Drug Administration (FDA) released a proposed rule that ends enforcement discretion for laboratory-developed tests (LDTs). The FDA's proposal outlines a five-stage implementation to begin regulating LDTs as they do for commercial in vitro diagnostics (IVDs), including modified FDA-approved/cleared tests. We outline here concerns from the clinical and public health microbiology laboratory perspective. It is our opinion that LDTs performed by individual Clinical Laboratory Improvement Amendments-certified diagnostic laboratories should not be regulated in the same way as commercial IVDs. This rule, if finalized, will negatively impact the diagnostic services currently offered by clinical and public health laboratories and, therefore, patients and the providers who care for them. Ending enforcement discretion will likely stifle diagnostic innovation and decrease access to diagnostic testing and health equity. Furthermore, the lack of infrastructure, including personnel and funding, at the FDA and diagnostic laboratories to support the required submissions for review is an obstacle. Like the FDA, diagnostic laboratories prioritize patient safety, accurate clinical diagnostics, and health equity. Since the scope of the LDT landscape is currently unknown, we are supportive of a registration process, along with non-burdensome adverse event reporting, to first understand the scope of clinical use of LDTs and any associated safety concerns. Any regulatory rule should be based on data that have been gathered systematically, not anecdotes or case reports. A rule must also balance the potential negative impact to patient care with realistic safety risks for infectious disease diagnostics.

Detection of a novel gross deletion in the UNC13D gene ends the diagnostic odyssey for a family with familial hemophagocytic lymphohistiocytosis 3

BackgroundFamilial hemophagocytic lymphohistiocytosis (FHL) is an immunological disorder characterized by overactivation of macrophages and T lymphocytes. This autosomal recessive condition has been characterized into multiple types depending on the genetic etiology. FHL type 3 is associated with bi-allelic pathogenic variants in the UNC13D gene.Case presentationWe present a 12-year diagnostic odyssey for a family with FHL that signifies the advances of FHL genetic testing in a clinical genetic diagnostic laboratory setting. We describe the first case of a large UNC13D gross deletion in trans to a nonsense variant in a family with FHL3, which may have been mediated by Alu elements within introns 12 and 25 of the UNC13D gene.ConclusionsThis case highlights the importance of re-evaluating past genetic testing for a patient and family as test technology evolves in order to end a diagnostic odyssey.

Strengthening clinical bacteriology laboratory diagnostics to combat sepsis and antimicrobial resistance in Benin: a train-the-trainer approach.

Improved laboratory diagnostics is needed to support sepsis diagnosis and combat increasing antibiotic resistance in Benin. We trained clinical laboratory experts and technicians to improve their skills in accurate and up-to-date diagnostics. A Train-the-Trainer (TtT) approach was used to design the course that combines theoretical and practical laboratory skills, specifically addressing the knowledge gaps we had previously identified in our national survey. Pedagogical methods were student-centered, including peer learning, use of online materials, practical laboratory work and pre-and post-course tests. We first trained 10 trainers who in turn trained 40 laboratory technicians from across the country, from both public and private clinical and veterinary laboratories. The trainers also prepared standard operation procedures for blood culture and antibiotic susceptibility testing based on international standards. Three months after the training, follow-up visits were made to the laboratories where the implementation of the new skills was evaluated. The progress of the participants observed during the course and the implementation of the new skills afterwards proved the training to be effective. The professional networks created during the training, the empowerment that utilizes local knowledge resources, and the government support for our initiative can be expected to bring sustainability to the initiative and support the participation of Beninese laboratories in international surveillance programs in the future.

Trace elements and the inflammatory process development: predictive possibilities

Introduction. Late diagnosis of inflammatory pathologies leads to increased risks of chronicization of the process, generalization, development of complications, and reduced effectiveness of therapy. Routine methods of clinical laboratory diagnostics often have diagnostic value at the stage of already developed disease with pronounced clinical manifestations. Determination of blood serum trace elements may have prognostic value in the diagnosis of inflammatory diseases. The trace elements most extensively studied in the context of inflammation and immune defense are copper and zinc. The aim of the study. The aim of the study was to investigate the prognostic significance of determining copper and zinc concentrations in serum in the diagnosis of inflammation. Material and methods. Concentrations of CRP, ferritin, ceruloplasmin, leukocytes, neutrophils, fibrinogen, copper, and zinc were determined in 1153 examined people aged 18 to 86 years. Serum trace elements were determined by ICP-MS method, the rest of the parameters were measured by standard methods. ROC analysis was used to evaluate the prognostic significance of serum copper and zinc measurements. Positive predictive value and negative predictive value were also calculated for the laboratory tests. Results. It was shown that serum copper concentration in both men and women can be used as a predictor of abnormalities of CRP, ceruloplasmin, and fibrinogen. By abnormalities in serum copper concentration, it is possible to predict increased leukocytes in men and women; decreased leukocytes in men; and increased neutrophil levels in men and women. The prognostic significance of the laboratory test for serum zinc was found in relation to the detection of ferritin deficiency in women and ceruloplasmin deficiency in men and women. Conclusion. The results obtained in the study suggest the possible use of serum copper and zinc laboratory tests for prognostic purposes.

Clinical Diagnostic Immunology in A Clinical Microbiology Laboratory

This review unravels the novel approaches, innovations and applications of immunology in the diagnosis of infections in a clinical microbiology laboratory. Many assays or tests in immunology are used in the diagnosis of infections. The availability of monoclonal antibodies has brought rapid progress to this area and they are used , for example , to identify specific microorganisms in culture or in tissues and fluids from infected patients. Examples of common diagnostic applications of antibody- antigen reactions discussed here includes agglutination tests, complement tests, complement fixation tests, ELISA, cytotoxic tests, radioimmunoassay tests, haemagglutination inhibition tests, hypersensitivity tests, phagocytic cell activity test, migration inhibition test, lymphocyte transformation test , lymphocyte fixation test , skin tests, delayed hypersensitivity tests and many others. The development of monoclonal antibody technology and the subsequent availability of unlimited quantities of pure antibody has improved the selectivity of these assays. The development of simple and highly sensitive methods for detection of nucleic acids is being applied to diagnosis of infection using nucleic acid hybridization and this provides a valuable additional aid to immunoassays in the clinical microbiology diagnostic laboratory.

ChatGPT Performance in Diagnostic Clinical Microbiology Laboratory-Oriented Case Scenarios.

Artificial intelligence (AI)-based tools can reshape healthcare practice. This includes ChatGPT which is considered among the most popular AI-based conversational models. Nevertheless, the performance of different versions of ChatGPT needs further evaluation in different settings to assess its reliability and credibility in various healthcare-related tasks. Therefore, the current study aimed to assess the performance of the freely available ChatGPT-3.5 and the paid version ChatGPT-4 in 10 different diagnostic clinical microbiology case scenarios. The current study followed the METRICS (Model, Evaluation, Timing/Transparency, Range/Randomization, Individual factors, Count, Specificity of the prompts/language) checklist for standardization of the design and reporting of AI-based studies in healthcare. The models tested on December 3, 2023 included ChatGPT-3.5 and ChatGPT-4 and the evaluation of the ChatGPT-generated content was based on the CLEAR tool (Completeness, Lack of false information, Evidence support, Appropriateness, and Relevance) assessed on a 5-point Likert scale with a range of the CLEAR scores of 1-5. ChatGPT output was evaluated by two raters independently and the inter-rater agreement was based on the Cohen's κ statistic. Ten diagnostic clinical microbiology laboratory case scenarios were created in the English language by three microbiologists at diverse levels of expertise following an internal discussion of common cases observed in Jordan. The range of topics included bacteriology, mycology, parasitology, and virology cases. Specific prompts were tailored based on the CLEAR tool and a new session was selected following prompting each case scenario. The Cohen's κ values for the five CLEAR items were 0.351-0.737 for ChatGPT-3.5 and 0.294-0.701 for ChatGPT-4 indicating fair to good agreement and suitability for analysis. Based on the average CLEAR scores, ChatGPT-4 outperformed ChatGPT-3.5 (mean: 2.64±1.06 vs. 3.21±1.05,P=.012, t-test). The performance of each model varied based on the CLEAR items, with the lowest performance for the "Relevance" item (2.15±0.71 for ChatGPT-3.5 and 2.65±1.16 for ChatGPT-4). A statistically significant difference upon assessing the performance per each CLEAR item was only seen in ChatGPT-4 with the best performance in "Completeness", "Lack of false information", and "Evidence support" (P=0.043). The lowest level of performance for both models was observed with antimicrobial susceptibility testing (AST) queries while the highest level of performance was seen in bacterial and mycologic identification. Assessment of ChatGPT performance across different diagnostic clinical microbiology case scenarios showed that ChatGPT-4 outperformed ChatGPT-3.5. The performance of ChatGPT demonstrated noticeable variability depending on the specific topic evaluated. A primary shortcoming of both ChatGPT models was the tendency to generate irrelevant content lacking the needed focus. Although the overall ChatGPT performance in these diagnostic microbiology case scenarios might be described as "above average" at best, there remains a significant potential for improvement, considering the identified limitations and unsatisfactory results in a few cases.