LabMed

Dried blood spots (DBSs) are a practical tool for diagnosing infectious diseases, especially in remote or resource-limited settings. This study assessed the efficacy of DBS-based serological assays for syphilis screening. EDTA blood samples from 171 syphilis-seropositive and 122 seronegative individuals were used to prepare DBSs by spotting whole blood onto filter paper. After drying, 12 mm disks were punched, incubated overnight in buffered solution, and centrifuged. Syphilis serological screening was conducted using the Liaison^® Treponema Screen assay, Macro-Vue™ Reagin Plasma Rapid (RPR) card test, and Dual Path Platform (DPP) Syphilis Screen and Confirm test. The Liaison^® assay demonstrated 100% sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) with an optimized cut-off. The nontreponemal RPR test showed very low sensitivity (2.9%) on DBS but perfect specificity (100%). The DPP test for treponemal antibodies achieved high sensitivity (92.1%) and specificity (98.2%) with microreader adjustment. Visual reading of the DPP test had variable accuracy, with sensitivity reaching 100% but lower specificity (42.1%). Nontreponemal antibody detection by DPP showed moderate sensitivity and specificity. Although nontreponemal testing requires refinement, DBS testing combined with point-of-care tests like DPP holds promise for expanding syphilis screening accessibility and decentralization globally, particularly in resource-constrained environments. Full article

A 39-question survey targeting recent graduates was deployed by the American Society for Clinical Pathology (ASCP) to its membership nationwide by email. Participants were prompted to reflect on clinical educators from whom they had learned the most and least. This survey was open for approximately six weeks with 177 respondents. Participants included medical laboratory scientists (71.8%), medical laboratory technicians (21.8%), phlebotomists (4.5%), blood bank specialists (0.9%) and laboratory administration (0.9%). This paper focuses on three survey questions. The first question asked participants to reflect on clinical educators from whom they had learned the most and explain why. Themes included teaching ability (37.2%), engagement (25.6%), passion (18.6%) and knowledge (16.3%). The second question asked participants to reflect on educators they had learned the least from and explain why. Themes included teaching challenges (48.8%), disengagement (29.3%) and unprofessionalism (19.5%). The third question asked about barriers to clinical training. Main themes included staffing shortages (25.8%), COVID-19-related issues (12.9%) and work culture (12.9%). Little research has been published on the student perspective of clinical training in laboratory sciences. This research provides insight into what students consider helpful in their training and what hinders their learning. Full article

Timely reporting of microbiological results is critical for clinical decision-making, particularly in pediatric hospitals where delays can significantly impact outcomes. Despite advances in laboratory automation, workflow inefficiencies and resistance to change remain barriers to improvement in Latin America. This study aimed to evaluate the effect of implementing a Kaizen-based change management strategy on reducing turnaround time (TAT) in the microbiology laboratory of Hospital Roberto del Río, Santiago, Chile. We conducted a prospective, pre–post intervention study focusing on blood culture processing. The baseline period (July 2022) included 961 cultures processed with the BacT/ALERT^® 3D system. A Kaizen/LEAN intervention was designed, comprising workflow redesign, staff training, and installation of the BACT/ALERT^® Virtuo^® (bioMerieux, Marcy l’Etoile, France) continuous-loading blood culture system. The intervention engaged all technical and professional staff in a five-day Kaizen immersion, followed by eight months of monitoring. Outcomes were assessed by comparing TAT for positive blood cultures before and after implementation (June 2023, 496 samples). Statistical analysis was performed using the Mann–Whitney U test, with p < 0.05 considered significant. The intervention achieved a median reduction in TAT from 68.22 h (IQR 56.14–88.59) pre-intervention to 51.52 h (IQR 41.17–66.57) post-intervention, corresponding to a 24.48% improvement (p < 0.001), surpassing the 20% target. Time to preliminary Gram reporting also decreased, and workflow standardization enhanced staff productivity and culture validation frequency. Implementation of Kaizen principles in a pediatric microbiology laboratory significantly reduced blood culture TAT and improved workflow efficiency. Beyond technological upgrades, active staff engagement and structured change management were key to success. These findings support the applicability of Kaizen-based interventions to optimize laboratory performance in resource-constrained public healthcare systems. Full article

Acute respiratory infections (ARIs) continue to pose a major global health threat, particularly among vulnerable populations. These infections often present with similar clinical symptoms, complicating accurate diagnosis and facilitating unmonitored transmissions. Genomic surveillance has emerged as an invaluable tool for pathogen identification and monitoring of such infectious pathogens; however, its implementation is frequently limited by high costs. The widespread use of high-throughput sequencing during the COVID-19 pandemic has created an opportunity to repurpose existing genomic platforms for broader respiratory virus surveillance. In this study, we evaluated the feasibility of adapting the Illumina COVIDSeq assay—initially designed for SARS-CoV-2 whole-genome sequencing—for use with Influenza A/B, Respiratory Syncytial Virus (RSV), and Rhinovirus. Positive control samples were processed using two approaches for library preparation: four virus-specific multiple workflows and a combined rapid workflow. Both workflows incorporated pathogen-specific primers for amplification and followed the Illumina COVIDSeq protocol for library preparation and sequencing. Sequencing quality metrics were analysed, including Phred scores, read length distribution, and coverage depth. The study did not identify significant differences in genome coverage and genetic diversity metrics between workflows. Genome Detective consistently identified the correct species across both methods. The findings of this study demonstrate that the COVIDSeq assay can be effectively adapted for multi-pathogen genomic surveillance and that the combined rapid workflow can offer a cost- and labour-efficient alternative with minimal compromise to data quality. Full article

Atrial fibrillation (AF), the most prevalent cardiac arrhythmia, significantly elevates the risk of stroke and heart failure. The etiology of AF is complex and multifactorial, involving genetic predisposition, environmental risk factors, and their potential interactions. A previous genome-wide association study (GWAS) of AF in a Korean population has identified an association between the rs3737883 single-nucleotide polymorphism (SNP) in the PPFIA4 gene and an increased risk of AF. However, the association needs to be replicated in other populations. In this paper, we conducted a case–control association study including 724 AF cases and 1475 controls, and successfully validated the association between SNP rs3737883 with the risk of AF in a Chinese population (OR = 1.33 with an adjusted p was 2.83 × 10⁻¹¹). Given that the PPFIA4 variant has been reported to influence high-sensitivity cardiac troponin T (hs-cTnT) levels, we further investigated the relationship between rs3737883 and hs-cTnT in 48 AF patients. Notably, we observed that the risk allele was also associated with elevated hs-cTnT levels. Our findings provide further genetic substantiation for the association of rs3737883 with AF. These results suggest a potential association between the PPFIA4 gene variant, hs-cTnT levels, and AF risk, although further studies are needed to clarify the underlying mechanisms. Full article

Non-coding RNAs (ncRNAs) are critical regulators of gene expression, taking part in the modulation of multiple biological functions across a range of cell types. Initially dismissed as transcriptional noise, ncRNAs are now recognized for their significant roles in key cellular mechanisms, including differentiation, apoptosis, and proliferation, as well as their profound implications for the pathogenesis of numerous human diseases. Due to their remarkable stability, tissue-specific expression patterns, and abundance in body fluids, ncRNAs hold significant promise as non-invasive biomarkers for diagnosis, prognosis, and therapeutic monitoring. Furthermore, advances in RNA-targeted therapeutics have introduced novel strategies to modulate ncRNA activity, although challenges related to delivery efficiency, specificity, and clinical validation remain. This review comprehensively summarizes the classification, biogenesis, and molecular functions of ncRNAs, elucidates their involvement in health and disease, and evaluates their potential as clinical biomarkers and therapeutic targets. Additionally, it discusses the emerging technologies for RNA manipulation, including CRISPR-based RNA editing, that can advance ncRNA research and revolutionize ncRNA-based therapeutics. Full article

Quality assurance in a clinical laboratory is essential to ensure reliable, accurate and precise laboratory test results all the time. A hemostasis laboratory is an important part of a clinical laboratory setting in a hospital or a healthcare center, and clinical laboratory tests play a crucial role in diagnosis and management of conditions related to bleeding or clotting of diseased individuals. This review discusses all aspects of coagulation laboratory testing from pre-analytical, analytical, and post-analytical variables as part of daily quality assurance processes undertaken as well as the quality management process of assay validation and implementation in a laboratory prior to patient testing. The internal and external quality processes that drive a hemostasis laboratory will be discussed that shows a rigorous process in assurance of testing that is reliable and accurate every time, at all times. Full article

Cell population data (CPD) parameters generated by Sysmex XN-series analysers are promising biomarkers for a variety of disease states. Routine use of CPD parameters, however, will require extensive evaluation of potential pre-analytical variables that may affect reliability. At present, no information on the comparability of CPD parameters generated using different blood collection tubes is available. In this preliminary study, we evaluated the impact of four commonly used blood collection tubes—dipotassium (K₂) EDTA, tripotassium (K₃) EDTA, trisodium citrate, and lithium heparin—on the generation of CPD parameters in whole blood from a cohort of 10 healthy donors. We also evaluated the stability of the CPD parameters generated at 4 h post-collection. Statistically significant differences in the CPD were observed across all blood collection tubes: whole blood anticoagulated with K₃EDTA induced minimal biases and was comparable to whole blood anticoagulated with K₂EDTA at collection; however, whole blood anticoagulated with citrate and heparin were associated with more substantial and more widespread biases in several parameters with potential clinical relevance. Notably, the biases observed in whole blood anticoagulated with K₃EDTA increased in both number and magnitude at 4 h post-collection, whilst the CPD parameters generated with whole blood anticoagulated with K₂EDTA remained stable. Although further confirmatory investigations are required, these findings highlight the importance of anticoagulant selection, as well as the need for further pre-analytical research, to support the integration of CPD parameters generated by Sysmex XN-series analysers into routine diagnostic workflows. Full article

In recent years, antiviral therapy has proved crucial in the treatment of infectious diseases, particularly infections by highly variable viruses such as human immunodeficiency virus, hepatitis B, hepatitis C, SARS-CoV-2 or bacteria such as Mycobacterium tuberculosis. Under the effect of selection pressure, this variability induces mutations that lead to resistance to antiviral and antibacterial drugs, and thus to escape from treatment. The use of Advanced Biological Laboratories (ABL) assays technology combined with next-generation sequencing (NGS) and automatized software to detect majority and minority variants involved in treatment resistance has become a mainstay for establishing therapeutic strategies. The present study demonstrated high concordance between majority and minority subtypes and mutations identified in 15 samples across four NGS platforms: ISeq100 (Illumina (San Diego, CA, USA)), MiSeq (Illumina), DNBSEQ-G400 (MGI (Santa Clara, CA, USA)) and Mk1C MinION (Oxford Nanopore (Oxford Science Park, UK)). However, nanopore technology showed a higher number of minority mutations (<20%). The analysis also validated the pooling of microbiological samples as a method for detecting mutations and genotypes in viral and bacterial organisms, using the easy-to-use DeepChek^® bioinformatics software, compatible with all four sequencing platforms. This study underlines the constant evolution of microbiological diagnostic research and the need to adapt rapidly to improve patient care. Full article

Cardiac amyloidosis (CA) results from the deposition of either immunoglobulin light chain (AL) or transthyretin (ATTR) amyloid fibrils in the myocardium, causing restrictive cardiomyopathy and, if left untreated, can lead to early death. Advancements in non-invasive diagnostic modalities have led to an increased recognition of the disease. Monoclonal gammopathy plays a pivotal role in the diagnostic algorithm for CA, particularly in differentiating AL from ATTR. This review highlights the importance of monoclonal protein detection through serum protein electrophoresis, immunofixation electrophoresis, and serum free light chain assays as initial screening tools. However, these tests alone are insufficient for a definitive diagnosis due to the complexities associated with coexisting monoclonal gammopathies and the potential for false negative and positive results. Advanced imaging modalities, such as echocardiography, cardiac magnetic resonance, and nuclear scintigraphy, along with tissue biopsy, are crucial for confirming CA and accurately determining the CA subtype. Full article

Rapid, safe, and field-deployable molecular diagnostics are crucial for the effective management of infectious disease outbreaks, particularly those involving highly infectious pathogens, which can produce clinical symptoms similar to less infectious pathogens, thus raising potential biosafety concerns. In this study, we evaluated DNA/RNA Defend Pro (DRDP) buffer, a novel viral-inactivating transport medium designed to stabilize nucleic acids and allow direct PCR without nucleic acid extraction. To ensure critical qPCR parameters were not compromised by using DRDP, we conducted serial dilution tests using herpes simplex viruses 1 and 2 (HSV-1, HSV-2) and varicella-zoster virus (VZV), comparing DRDP to standard universal transport medium (UTM). Detection sensitivity, determined by cycle quantification (Cq) values, favored DRDP, as UTM samples required a 2–3-fold dilution to mitigate PCR inhibition. DRDP maintained reliable PCR compatibility at reaction volumes containing up to 25% buffer. At higher DRDP concentrations (30–35%), PCR inhibition occurred due to EDTA content but was fully reversible by adding supplemental magnesium. Furthermore, DRDP samples did not require an initial 95 °C thermal lysis step, thus simplifying the procedure without reducing PCR sensitivity or efficiency. Full article

Fluid-sensitive sequences on MRI [Magnetic Resonance Imaging] have widely been used to assess soft tissue oedema. Windowing techniques play a significant role in adjusting the contrast to highlight the pathology. The purpose of this study is to establish the impact of modified MRI window parameters, with a narrower window width than window level, in assessing soft tissue oedema in a plethora of musculoskeletal pathologies. Fifty randomly selected patients with a range of musculoskeletal pathologies resulting in soft tissue oedema on MRI were included in the study. Two separate images of each MRI study were taken on a PD fat suppressed sequence, one with default windowing range and another with window width lower than that of window level. Both images were reviewed by two radiologists and were assessed for diagnostic effectiveness in terms of image resolution and depiction of pathology. Assessment was semi-quantitatively compared and graded on the Likert scale, from 1 to 5, with 1 indicating poor quality and 5 indicating excellent quality. Friedman’s test was then conducted to compare the scores of both images. In most of the cases, the image with the modified window/level setting was significantly better in terms of depicting pathology and having better resolution, though some cases showed no clear preference. Friedman’s test showed that the score for images with modified window settings was significantly higher. Images with modified windowing in conjunction with standard imaging protocols help to assess soft tissue oedema. Full article

Direct oral anticoagulants (DOACs) have been licensed worldwide for several years for various indications. Each year, 10–15% of patients receiving oral anticoagulants will undergo an interventional procedure, and expert groups have issued several guidelines for perioperative management in such situations. According to the PAUSE study, the proposed randomized strategy of stopping DOACs without bridging therapy in patients with atrial fibrillation was associated with low rates of major bleeding and arterial thromboembolism so that its implementation is increasingly safe. The present study was carried out in order to investigate the efficacy and safety of the standardized perioperative DOAC management strategy by measuring the residual activity of oral anticoagulants when stopping them preoperatively in daily practice in a regional hospital. Thirty-two patients were included in the present study. They were patients who suffered from atrial fibrillation or deep vein thrombosis and were receiving an oral anticoagulant, rivaroxaban or apixaban at the indicated dose. These patients underwent an elective surgery or invasive procedure at the Karditsa General Hospital between May 2022 and April 2023. The results showed that in a percentage of >90% of the patients on the day of surgery they had a residual anti-Xa activity below 0.5 U/mL. This rate is considered high and confirms the safety and efficacy of the guideline-recommended protocol for perioperative discontinuation of DOACs. Full article

This study included deidentified antibiotic susceptibility results from outpatient urinary Escherichia coli isolates from Washington state which were tested at a large clinical laboratory during 2013–2019. Isolates were categorized as representing the first, second, third, or fourth-or-greater occurrence of infection in data from individual patients. We used logistic regression with the outcome of resistance, adjusting for year of antimicrobial susceptibility test, patient sex, patient age, and facility type. In cases of subsequent infection, we found a significant risk of resistance to levofloxacin, ciprofloxacin, ceftriaxone, trimethoprim-sulfa, nitrofurantoin, ampicillin, gentamicin, and amoxicillin-clavulanate. Our findings suggest that Escherichia coli isolates from recurrent urinary tract infections have a higher rate of resistance to most tested antibiotics than isolates from the first urinary tract infection in a given year. However, susceptibility frequencies did not differ significantly between antibiograms constructed using only the first occurrence in a patient and those constructed using all subsequent occurrences. These findings suggest that the traditional approach of including only the first occurrence of urinary Escherichia coli in a patient may underestimate levels of antibiotic resistance in a community. Such underestimation could negatively affect empiric therapeutic choices, health outcomes, and treatment costs. Full article

Objective: This study employed advanced biostatistical methods to investigate Interleukin-6 (IL-6), Tumour Necrosis Factor Alpha (TNF-α), and Myeloperoxidase (MPO) levels in serum and plasma samples from patients with severe osteoarthritis (OA) compared to volunteers. The primary aim was to evaluate the diagnostic potential of these biomarkers and address statistical challenges, including non-normal data distribution and non-aged-matched groups. Design: Using Enzyme-Linked Immunosorbent Assays (ELISAs), IL-6, TNF-α, and MPO concentrations were analysed in 58 OA patients and 28 volunteers. Statistical analyses included Shapiro–Wilk tests to assess normality, a Mann–Whitney U (MWU) test to compare biomarker levels, and sensitivity analyses using Rank-based ANCOVA, and regression models were used to address non-normal data distributions and to validate the findings under adjustments for age and gender. Levene’s test was used to evaluate the homogeneity of variables. Results: Serum TNF-α and plasma MPO were significantly higher in OA patients than in volunteers (p < 0.05), while IL-6 levels were non-significant (p = 0.160). MWU tests confirmed significant differences for TNF-α (p = 0.045) and MPO (p = 0.0001). Sensitivity analysis using Rank-based ANCOVA and regression models confirmed the robustness of these biomarkers, with TNF-α (p = 0.037) and MPO (p = 0.0099) retaining statistical significance after adjusting for covariates. IL-6 remained non-significant across all analyses. Conclusions: TNF-α and MPO emerged as statistically robust biomarkers for severe OA, with the serum samples better reflecting inflammation than plasma. These findings underscore the importance of using advanced biostatistical methods such as Rank-based ANCOVA and regression to validate biomarkers, particularly in heterogenous datasets. Future research should incorporate larger, more diverse cohorts and detailed demographic profiling to explore the early diagnostic potential of these biomarkers and further understand OA progression. Full article

Background: In vitro blood circulation loop systems are utilized to assess the hemocompatibility and performance of medical devices that come into contact with blood, in accordance with the international standards ASTM F1830 and ASTM F1841. However, a method for evaluating the specific type of anticoagulant and the blood characteristics of each animal species is necessary to ensure consistent and reliable results. Methods: Blood was collected from healthy rabbits, pigs, rhesus monkeys, and cynomolgus monkeys to evaluate whole blood preserved in anticoagulants (ACD-A, CPDA-1, and heparin). For each sample, red blood cells were monitored over time, and their morphological characteristics were documented. Results: The morphological grade of erythrocytes gradually decreased over time. Significant differences were observed based on the type of anticoagulant used in the experiment, and variations were noted among different animal species. Conclusions: The hemocompatibility of in vitro blood circulation loop systems may vary depending on the animal species. Observing erythrocyte morphology can serve as a control measure to ensure reproducible results. Full article

Identification of mycobacterial infections for both Mycobacterium tuberculosis and non-tuberculosis mycobacteria is important for effective patient care. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is a promising tool that is used in many clinical laboratories for the identification of bacteria and yeast. This study evaluates the impact of growth media on the performance of the MALDI Biotyper^® MBT smart MS for mycobacteria identification. Increased rates of identification, particularly in non-rapid growers and pigment producers, and higher confidence scores were generated in mycobacteria isolated from solid agar, rather than liquid broth. Testing each isolate in triplicate can increase yield of detection. Using the Bruker MBT Mycobacteria Kit to process our samples for testing on the Bruker MALDI Biotyper^® instrument generated precise and accurate mycobacteria identification. These findings emphasize the importance of optimizing mycobacterial specimen processing workflows to include appropriate culture media, which can enhance mycobacterial identification and improve diagnostic accuracy. Full article

The objective of this study was to assess the performance characteristics of the new automated haematology analyser from Sysmex Corporation, the Sysmex XR-Series, compare its performance to the Sysmex XN-Series through method comparison, and compare our results to previously published literature. Analytical performance of the new Sysmex XR-20 consisting of precision, bias, and total error, a method comparison with the Sysmex XN-2000, and the flagging performance evaluation were conducted on a Sysmex XR-20 analyser in the AZ Sint-Lucas Hospital (Bruges, Belgium) several months before its launch in Europe. We conclude that the Sysmex XR-Series is an excellent successor to the Sysmex XN-Series for routine haematology analysis. Analytical performance and flagging efficiency are comparable to the Sysmex XN-analyser. Full article

Cell-free DNA (cfDNA) analysis is a pivotal tool in non-invasive diagnostics, including cancer monitoring and prenatal testing. However, the preanalytical phase, particularly the choice of anticoagulant, significantly impacts cfDNA integrity and yield. This study aims to compare cfDNA yield, stability, and DNase activity in plasma-citrate and plasma-heparin, using plasma-EDTA and serum as established controls, to explore more deeply the impact of blood DNAse activity on cfDNA in these specimens. Blood samples from 15 healthy volunteers were collected in four types of tubes (citrate, heparin, EDTA, and serum). cfDNA was extracted and quantified using qPCR, and endogenous DNase activity was assessed through hydrolysis probe assays. Samples were incubated at 37 °C for 24 h to evaluate cfDNA degradation rates. Heparin-plasma exhibited the highest DNase activity, with baseline cfDNA levels intermediate—higher than EDTA but lower than serum—leading to substantial cfDNA degradation (85.3%). Combined with its known PCR inhibition, this renders heparin-plasma unsuitable for cfDNA analysis. Citrate-plasma, with baseline cfDNA levels similar to EDTA, showed partial DNase inhibition, resulting in intermediate cfDNA degradation (13.3%), a limitation that diminishes its viability compared to EDTA-plasma. Serum, with the highest baseline cfDNA levels, exhibited high DNase activity and significant cfDNA degradation (55.6%), making it unsuitable for cfDNA preservation. EDTA-plasma demonstrated complete DNase inhibition and minimal cfDNA degradation (8%), confirming it as the most suitable specimen for cfDNA analysis. These findings emphasize the importance of anticoagulant selection, highlighting critical limitations of heparin-plasma and citrate-plasma while reinforcing EDTA-plasma as the gold standard for preserving cfDNA integrity in diagnostic applications. Full article