LabMed, Volume 2, Issue 1 (March 2025) – 4 articles

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