LabMed

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

Recent initiatives have discouraged the treatment of asymptomatic bacteriuria in specific patient populations due to its lack of clinical benefit, no improvement in morbidity or mortality, and its contribution to antibiotic overuse. This study aimed to evaluate whether an intervention at order entry, combined with DCLS laboratory consultation for urine cultures and urinalyses, could reduce unnecessary lab tests and inappropriate antibiotic use, thereby improving patient outcomes. Our research design was a quasi-experimental study with a retrospective and prospective chart review on non-pregnant adult patients 18 years of age and older from July 2021 to September 2022. Data collected for both reviews included patient demographics, provider demographics, patient signs and symptoms, laboratory test results, test order type, test order utilization and antibiotic prescriptions. Our study included 6372 patients, with 3408 in the retrospective review and 2964 in the prospective review. Before the intervention, 60% (n = 2053) of test orders were inappropriate, which decreased to 20% (n = 591) post-intervention. In asymptomatic patients, reflexed urine cultures decreased from 51% to 13% post-intervention. Lastly, in asymptomatic patients, antibiotic therapy at discharge dropped from 54% to 25% after the intervention. Post-intervention ordering practices improved, decreasing the number of inappropriate orders across all patient and provider types. Overall, this initiative showed a significant reduction in the treatment of asymptomatic bacteriuria, which has been linked to the overuse of antibiotic therapy. Full article

Acute promyelocytic leukemia (APML) is one of the most curable leukemia subtypes, where the majority of patients achieve complete remission and also deep molecular remission after therapy, characterized by a PCR-undetectable state. Similarly, chronic myelogenous leukemia (CML) is a leukemia where, thanks to effective targeted treatment with tyrosine kinase inhibitors (TKIs), deep remission detectable only by PCR has become part of the routine management of these patients. Here, we describe a patient who was PCR-negative after induction and consolidation with arsenic trioxide (ATO) and all-trans retinoic acid (ATRA) and stayed PCR-undetectable for 13 months post-consolidation, later experiencing molecular relapse following mild SARS-CoV-2 infection. The patient was able to reestablish molecular remission again without anti-leukemic therapy several weeks later. She remained PCR-negative for the next 42 months. Viral infection-triggered immunosuppression, as in our case, offers a possible explanation for the temporary loss of molecular remission seen in leukemia patients monitored by PCR. Our first case illustrates this period of convalescence from viral infection, which was maybe accompanied by loss of molecular response. Viral infections and temporary immunosuppression may be a culprit in cases where molecular responses are lost temporarily. This loss of the PCR-undetectable state may have implications for other cancer patients where PCR monitoring is used. Thus, our observation may have broader implications for other patients, especially those with CML. We further enforce these findings by describing a second patient with CML who experienced temporary molecular relapse in the setting of post-viral syndrome. Full article

Sepsis is a life-threatening condition, and clinicians should diagnose it as soon as possible to enable rapid intervention. The study aims to validate the AIA-360 Presepsin (PSEP) test for its use in determination in the diagnosis of septic patients after admission to emergency departments (ED). A total of 97 blood samples were collected from patients at the ED and from blood donors of Tor Vergata Hospital. Here, 15 samples were obtained from patients with a confirmed diagnosis of sepsis, and 44 samples with non-septic inflammatory condition. A control group of 38 samples from healthy subjects was also included. The non-septic inflammatory condition group and the confirmed sepsis group had a median of 874.40 pg/mL and 1467.10 pg/mL, respectively, while the control group showed a PSEP median value of 473.90 pg/mL, thus showing a significant statistical difference among all groups. The ROC curves highlighted a good sensitivity (93.33%) and specificity (76.19%) for PSEP values, suggesting the best cut-off point of 890 pg/mL. (p-value < 0.001; Mann–Whitney test). The PSEP test can improve and speed up the diagnosis of sepsis after admission to the ED with respect to other biomarkers, mainly due to its early kinetics. Full article

Anticoagulants can complicate the interpretation of routine and specialised coagulation assays. Several methodologies have been developed to minimise or eliminate anticoagulant-associated interferences; however, no ‘universal methodology’ that encompasses different anticoagulant classes is currently available. Ciraparantag is a promising reversal agent that can bind both direct oral anticoagulants (DOACs) and heparin-like anticoagulants. As such, we aimed to investigate whether ciraparantag could be employed as a ‘universal’ anticoagulant chelator in vitro. Human plasma was spiked with ascending concentrations of ciraparantag, with or without DOACs or heparin, and assayed for routine coagulation parameters. Ciraparantag had minimal effects on coagulation testing when added to human plasma at concentrations similar to pharmacokinetic maxima; however, ciraparantag did not remove DOAC- or heparin-associated activities in vitro, which was likely due to the preferential chelation of anionic substances in the coagulation reagents. In contrast, DOAC-Stop™, a commercial activated charcoal-based adsorbent, efficiently removed both DOAC- and ciraparantag-associated interferences. In conclusion, although ciraparantag is not effective as a ‘universal’ anticoagulant chelator in vitro, we report that activated charcoal-based adsorbents may be clinically useful in situations where laboratory investigations are complicated by the presence of DOACs and/or ciraparantag. Full article

In this study, we present a novel face mask engineered for the collection of exhaled breath condensate (EBC) and its application and performance in a clinical study of COVID-19 infection status assessment versus the gold standard polymerase chain reaction (PCR) nasopharyngeal swab testing. EBC was collected within a clinical trial of COVID-19-infected and non-infected patients and analyzed by reverse transcription quantitative (RT-q) PCR, with the results being compared with nasopharyngeal sampling of the same patient. The cycle threshold (Ct) values of the nasopharyngeal samples were generally lower than those of EBC, with viral loads in EBC ranging from 1.2 × 10⁴ to 5 × 10⁸ viral particles mL⁻¹ with 5 min of breathing. From the 60 clinical patients’ samples collected, 30 showed a confirmed SARS-CoV-2 infection. Of these 30 individuals, 22 (73%) had Ct values < 40 (representing the threshold for SARS-CoV-2 infectivity) using both amplification of ORF1a/b and the E-gene. The 30 EBC samples from non-infected participants were all identified as negative, indicating a 100% specificity. These first results encourage the use of the face mask as a noninvasive sampling method for patients with lung-related diseases, especially with a view to equipping the face mask with miniaturized sensing devices, representing a true point-of-care solution in the future. Full article

Quantifying M-proteins is an important part of diagnosing and monitoring patients with monoclonal gammopathies. Historically, laboratories use one of two methods to accomplish this. The splice method utilizes a perpendicular drop on each side of the M-protein on the electrophoretogram. In contrast, the skim method applies a tangent skimming line connecting the points above the polyclonal background. In this study, we compared the bias between these two methods across two different instruments (Helena SPIFE 3000 and Sebia Capillarys 3) in 118 patients. First, we compared the splice technique on both instruments and observed a significant average bias of 58.3% (slope = 1.437, y-intercept = 0.76, and r = 0.9682). We next compared the splice technique on the SPIFE 3000 to the skim technique on the Capillarys 3 and observed an average bias of only −2.10% (slope = 1.363, y-intercept = −1.98, and r = 0.9716), although there was significant scatter along the line of best fit. Lastly, we compared splice vs. skim on the Capillarys 3 and observed an average bias of −38.2% (slope = 0.947, y-intercept = −2.65, and r = 0.9686). Based on these results, care should be taken when switching instruments or integration techniques to ensure consistent monitoring of patients. Full article

The assessment and modelling of DNA double-strand break damage and repair is widely investigated throughout the literature. This optimisation study investigated the requirement of cell proliferation prior to treatment with chemotherapeutic agents to damage DNA and the optimal window of analysis for DNA double-strand break repair measurements with γ-H2AX. Peripheral blood mononuclear cells were collected from healthy volunteers and incubated with phytohaemagglutinin at final concentrations of 0, 0.25, 0.5, 1, 2.5, 5 and 10 µg/mL for 0, 24, 48, 72 and 168 h at 37 °C, 5% CO₂, and proliferation was measured via spectrometry (MTS assay). This study, detailed in this methodology paper, found that peripheral blood mononuclear cells must be proliferated prior to the chemical induction of DNA double-strand breaks. The window for assessment of early DNA double-strand break repair was determined to be one hour after removal of the DNA damaging agent. Full article