Bloodstream infections (BSIs) carry high morbidity and mortality, and delayed targeted therapy worsens survival. Conventional antimicrobial susceptibility testing (AST) from positive blood cultures typically requires 24–48 h (and >72 h for some resistant organisms), delaying appropriate treatment and leaving 14–78% of empirical regimens unnecessary or ineffective. Direct rapid antimicrobial susceptibility testing (RAST) addresses this bottleneck by delivering AST in 4–16 h with categorical agreement (CA) ≥90% for major pathogens, using volatile organic compound (VOC) detection, modified disk diffusion, microfluidics, and molecular approaches. RAST generally makes susceptibility results available approximately 8–44 h earlier, with savings exceeding 56 h when conventional testing requires more than 72 h. This review compares the principles, turnaround times (TAT), and performance of leading RAST platforms. It further identifies persistent bottlenecks restricting cross-platform comparability and clinical uptake, most notably the absence of standardized protocols, the inability of molecular assays to determine minimum inhibitory concentrations (MICs), and unresolved pre-analytical confounders. Although RAST reliably shortens time-to-result and supports antimicrobial stewardship, its routine integration demands platform-specific validation, transparent regulatory and cost-effectiveness assessment.
Background The growing demand for point-of-care testing (POCT) has increased interest in at-home blood testing. However, practical methods for obtaining serum from small-volume blood samples remain limited. This study investigated the effects of dilution buffers and additives on RBC aggregation and clot-formation behavior in diluted blood to develop a novel serum extraction method. Methods Residual blood samples obtained from outpatients were used to evaluate RBC aggregation and clot-formation behavior with various dilution buffers and additives, including polyethylene glycol (PEG). Based on effective combinations, a filtration-based method for serum extraction from small-volume blood samples was developed and evaluated. Results Dilution with saline or phosphate-buffered saline resulted in large clots and numerous floating red blood cells (RBCs). In contrast, the high-molecular-weight PEG reduced clot size and markedly decreased the number of floating RBCs. Hypertonic dilution buffers also reduced floating RBCs without disrupting clot formation. By combining PEG with hypertonic buffers, serum was successfully extracted by manual filtration without hemolysis. The extracted serum enabled C-reactive protein measurement that strongly correlated with measurements obtained from conventional serum samples. Conclusions A simple and rapid method for hemolysis-free serum extraction from small-volume blood samples was developed using PEG and hypertonic buffers. This approach may facilitate the development of POCT applications requiring reliable serum collection from limited blood volumes.
This article reports a clinical case of falsely and significantly elevated high-sensitivity cardiac troponin I (hs-cTnI) levels caused by “Macro-Troponin I". A 54-year-old male patient was admitted with a diagnosis of acute non-ST-segment elevation myocardial infarction (NSTEMI). While his hs-cTnI levels rose sharply during the acute phase and subsequently persisted at an extremely high plateau level (>27404 ng/L) without decline for several days, his myoglobin (MYO) and creatine kinase-MB (CK-MB) levels followed the expected pattern of initial elevation followed by resolution. This discrepancy raised suspicion regarding the validity of the hs-cTnI results. Subsequent systematic laboratory verification, including multi-platform assay comparison revealing a vast discrepancy between hs-cTnI and high-sensitivity cardiac troponin T (hs-cTnT) results, non-linear recovery upon serial dilution, and a marked decrease in hs-cTnI to normal levels after polyethylene glycol (PEG) precipitation, confirmed interference from a precipitable macromolecular complex. Furthermore, the interference could not be eliminated by various common heterophilic antibody blocking agents.Based on laboratory results, we confirmed the presence of a macromolecular interference in the patient's plasma, which was highly likely to be macro cardiac troponin I due to its ultra-long half-life.Regarding the origin of macro cardiac troponin I, current research indicates that anti-troponin antibodies exist, which rapidly form immune complexes upon exposure to troponin antigens, thereby interfering with experimental data.Through this case, we demonstrated the presence of macro-complexes interference. The patient ultimately received correct diagnosis and management due to timely recognition of this interference.
HbA1c and blood glucose (BS) are routinely used in diabetic care, but they do not provide direct information about ongoing cellular pathogenic processes. Previous studies in diabetic animal models identified a CD106+TNFα+proinsulin+ subset within short-term hematopoietic stem/progenitor cell populations, termed diabetes stem cells, that was implicated in diabetic pathology. In this study, we examined peripheral blood from patients with type 2 diabetes mellitus (T2DM) (n = 12) and healthy volunteers (n = 10). The proportion of CD106+TNFα+proinsulin+CD34+ cells was significantly higher in patients with T2DM than in healthy controls and showed a significant positive correlation with HbA1c levels. These findings suggest that this aberrant peripheral blood CD34+ subset may represent a candidate cellular biomarker associated with T2DM and chronic glycemic status. Further studies in larger cohorts are required to evaluate its clinical utility.
Background:Thyroid-stimulating hormone (TSH) is the primary biomarker for diagnosing and monitoring thyroid disorders. Although serum remains the reference matrix, microsampling approaches, including dried blood spots (DBS) and volumetric devices (CPT B50), may improve accessibility and compliance. This study evaluated the analytical and clinical performance of TSH measurement from DBS and CPT B50 compared with serum. Methods:Paired serum, DBS, and CPT B50 samples were collected from 246 adults undergoing thyroid evaluation. DBS and CPT B50 samples were eluted and analyzed using a chemiluminescence immunoassay (RUO, LIAISON® XL, DiaSorin). Analytical performance, recovery, repeatability, stability, and method agreement were assessed. Microsample results were converted to serum-equivalent concentrations using inverse Passing-Bablok regression equations. Performance was evaluated using Passing-Bablok regression and Bland-Altman analysis with predefined acceptance criteria (bias ≤ ±15%, CV ≤ 15%, recovery 85%-115%), guided by CLSI EP15-A3 and EP17-A2 guidelines. Results:Repeatability CVs ranged from 3.7% to 10.4%, and recovery was 97%-110%. DBS and CPT B50 showed strong agreement with serum, with mean bias of -0.10 mIU/L for CPT B50 and 0.24 mIU/L for DBS. Samples remained stable for up to 8 weeks at +4 °C and -20 °C. Thyroid status classification accuracy was 96% for DBS and 98% for CPT B50. Conclusions:TSH measurement from DBS and CPT B50 showed high analytical and clinical concordance with serum. Volumetric sampling improved reproducibility and stability, supporting its use in remote thyroid testing and therapy monitoring.
A lactate concentration ≥4 mmol/L is associated with organ dysfunction and poor outcomes in sepsis, making accurate measurement and timely communication of critical results vital. Photometric lactate assays are common but prone to spectral interference from icterus, and objective assessment of haemolysis/icterus/lipaemia indices is essential to prevent release of compromised results. The authors report a patient with severely elevated lactate (10.2 mmol/L) in fluoride-oxalate plasma. The specimen was run on a colourimetric lactate oxidase method (Beckman Coulter AU5800 analyser at Cork University Hospital (CUH)). Since icteric indices were not routinely determined for fluoride-oxalate specimens, the sample was assessed by eye to be grossly icteric, and the initial decision was to withhold the lactate result per manufacturer guidelines. On review by the Consultant Clinical Biochemist, an automated icteric index was requested, which yielded a 1+ flag, corresponding with a total bilirubin in the range 42.7-85.5 μmol/L, and inconsistent with gross icterus. Review of the patient history indicated aplastic anaemia. Drug-related discolouration of the sample was suspected, and the haematology team confirmed eltrombopag therapy. The lactate result and subsequent reports were released with advisory comments, noting that the degree of any eltrombopag-related analytical interference was uncertain. This case demonstrates how a combination of drug-induced plasma discolouration, reliance on visual assessment of icterus, and inconsistent application of HIL indices across specimen types jeopardises the release of critical lactate results. Standardised automated index testing, clear protocols for interference evaluation, and timely multidisciplinary discussion of medication history are important safeguards in clinical laboratory practice.
Objective This study aimed to systematically evaluate the interference of 49 commonly used clinical drugs on 60 routine biochemical tests, providing a foundation for clinical laboratories to establish drug-interference alert systems and support more accurate result interpretation. Method We tested 49 drugs at three concentrations (baseline, therapeutic C1, supra-therapeutic C2) on 60 biochemical assays following CLSI EP07. For each drug-test combination, a single patient serum/plasma pool was split into three aliquots: baseline, C1-spiked, and C2-spiked. All samples were analyzed using a Mindray BS-2800M fully automated biochemical analyzer. Results at C1 and C2 were compared with baseline (zero concentration) to characterize drug-induced interference with each biochemical test. Results At concentration C1, 23 drugs exhibited no interference across all 60 biochemical tests. Among the 2940 drug-test combinations evaluated, C1 generated 27 cases (0.9%) of strong interference, mainly within liver function assays, and 40 cases (1.4%) of weak interference. The majority (97.7%) exhibited no detectable effect. At concentration C2, a total of 2820 drug-test combinations were included in the analysis. Strong interference was detected in 103 cases (3.7%), primarily affecting liver and lipid profiles, alongside 117 cases (4.1%) of weak interference. Nonetheless, 92.2% of combinations still showed no interference. Conclusion This study provides a comprehensive profile of drug-induced interference across 60 routinely performed biochemical tests. These findings could support the accurate interpretation and interference identification of laboratory results.
Objectives:To establish age-specific reference intervals for serum copper in healthy Vietnamese children and adolescents using a data-driven age-partitioning approach and to assess its implications for result interpretation. Methods:Apparently healthy participants from birth to <19 years were recruited. Serum copper was measured by graphite furnace atomic absorption spectrophotometry. Candidate age thresholds were explored using recursive partitioning analysis and subsequently rounded to clinically interpretable groups. Final age partitions were determined based on recursive partitioning results, between-group statistical comparisons, effect size, clinical interpretability, and sample size. Differences between adjacent age groups and between sexes were assessed using the Mann-Whitney U test, Cliff's delta, and the Harris-Boyd method. Outliers were identified using the Horn algorithm. Reference intervals were estimated by the nonparametric percentile method in accordance with CLSI EP28-A3c. Results:Of 1157 eligible participants, 36 outliers were excluded, leaving 1121 for analysis. Final age groups were 0-<6 days, 6 days to <12 months, 12 months to <7 years, and 7 to <19 years. Significant differences were observed between adjacent age groups, supporting age partitioning. Sex-related differences were small and did not support sex-specific partitions. The derived reference intervals were 1.9-15.4 μmol/L, 6.5-26.9 μmol/L, 9.3-35.8 μmol/L, and 7.9-22.6 μmol/L for the respective age groups. Conclusions:Serum copper concentrations vary substantially with age, particularly in early life, while sex differences are limited. Age-specific, but not sex-specific, reference intervals are recommended. These findings support clinically appropriate age partitioning to improve interpretation of pediatric copper results and reduce potential misclassification associated with overly broad age groups.
Objective To investigate the distribution characteristics of potential high-risk pathogens for early-onset neonatal infection in maternal vaginal secretions, and to perform a head-to-head comparative evaluation of detection performance for target pathogens between metagenomic next-generation sequencing (mNGS) and real-time quantitative polymerase chain reaction (qPCR), with conventional bacterial culture as the reference standard. Methods A total of 294 valid maternal vaginal secretion samples were prospectively collected and tested in parallel using qPCR, mNGS, and conventional bacterial culture. The Chi-square test was used to compare the differences in pathogen detection rates among the three methods. Receiver operating characteristic (ROC) curve was plotted to calculate the area under the curve (AUC) and 95% confidence interval (CI), to systematically evaluate the detection performance of the two methods for target pathogens. Results The spectrum of potential early-onset neonatal pathogens in maternal vaginal secretions, ranked by detection rate, was as follows: Staphylococcus aureus, Streptococcus agalactiae, Ureaplasma urealyticum, Listeria monocytogenes, and Campylobacter fetus. The detection rates of these target pathogens by qPCR, mNGS, and bacterial culture showed high consistency, with no statistically significant difference in detection rates among the three methods (all P > 0.05). ROC curve analysis showed that the AUC values of both qPCR and mNGS for the above major pathogens were all above 0.90, which were significantly different from the null hypothesis of AUC = 0.5 (all P < 0.05), indicating good detection performance; while there was no significant difference in AUC values between qPCR and mNGS (all P > 0.05). In addition, Listeria monocytogenes (3 cases) and Campylobacter fetus (1 case) were only detected by qPCR and mNGS, while not isolated by conventional culture. Conclusion This head-to-head comparative study confirms that both mNGS and targeted qPCR have high accuracy and consistency for detecting potential early-onset neonatal pathogens in maternal vaginal secretions. We propose a tiered antenatal screening strategy for maternal vaginal pathogenic colonization: qPCR is recommended as the first-line tool for routine antenatal screening due to its high cost-effectiveness and rapid turnaround time, while mNGS is reserved for high-risk pregnant women (e.g., preterm premature rupture of membranes, clinical chorioamnionitis), culture-negative suspected infection cases, or scenarios requiring comprehensive pathogen profiling, to take full advantage of its unbiased, broad-spectrum detection capability. This integrated screening strategy requires further prospective validation with paired neonatal clinical outcome data to confirm its value in the prevention and early intervention of early-onset neonatal infection.
Introduction Quality management systems are essential in clinical laboratories to ensure optimal operational output. However, report generation still frequently relies on manual processes which are time-consuming and prone to errors. Methods A rule-based artificial intelligence tool was internally developed to automate quality management report generation by directly extracting and processing electronic laboratory records from the health information system using pre-defined formulas and logic. Results Implementation of this tool in a Medical Genetics laboratory reduced report preparation time by 90% and eliminated discrepancies compared to manual reports, alleviating the need for extensive secondary reviews. Conclusion This AI-assisted approach streamlines quality management reporting, enhancing efficiency and data consistency. The successful development and implementation of those tools require continuous communication and validation between the different stakeholders for effective system refinement.
Background:Accurate BCR::ABL1 quantification is essential for monitoring chronic myeloid leukemia (CML), particularly at the major molecular response (MMR; ≤0.1% IS) and deep molecular response (DMR) levels: MR4 (≤0.01% IS) and MR4.5 (≤0.0032% IS). We evaluated the analytical performance of the 1copy BCR::ABL1 qPCR Kit (1drop, Republic of Korea), a one-step real-time quantitative PCR (qPCR) Kit and compared it with two established assays. Methods:Precision, linearity, and detection capability (LoB, LoD, LoQ] were assessed according to Clinical and Laboratory Standards Institute (CLSI) guidelines. A method comparison study was performed using 74 clinical samples tested concurrently with the ipsogen BCR::ABL1 Mbcr IS-MMR Kit (Qiagen, Hilden, Germany) and the QXDx BCR::ABL %IS droplet digital PCR (ddPCR) assay (Bio-Rad, Hercules, CA, USA). Concordance was further evaluated at clinically critical molecular response thresholds (>10%, >1%, and ≤0.1% international scale [IS]). Results:The coefficients of variation at 14.1% IS and 0.233% IS were 14.4% and 15.6%, respectively. The assay was linear across a 5-log range (0.02-85% IS; R2 = 0.9949). The manufacturer-claimed LoB, LoD, and LoQ were verified. Pearson correlation coefficients with the ipsogen and QXDx assays were 0.951 and 0.950, respectively. Concordance at the >10% IS, >1% IS, and ≤0.1% IS thresholds was 98.6%, 100%, and 95.9% (vs. ipsogen), and 100%, 100%, and 90.5% (vs. QXDx), respectively. Conclusions:The 1copy BCR::ABL1 qPCR assay demonstrated reliable analytical performance and high concordance with established quantitative assays at clinically relevant molecular response thresholds, supporting its utility for routine monitoring of CML.
Objective The high cost of imported platforms limits widespread lung cancer risk stratification in resource-limited settings. Based on CLSI guidelines, this study aimed to conduct a preliminary analytical verification of a novel automated chemiluminescence immunoassay system (Health) for five tumor markers (SCC, CEA, CYFRA21-1, Pro-GRP, and NSE), and to compare it with established reference systems. Methods Analytical precision and method comparison with Roche, Abbott, and Beckman platforms were verified referencing CLSI EP recommendations. A total of 180 patients with lung cancer and 77 healthy controls were enrolled. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic performance of individual tumor markers and their combined use. Results The evaluated system demonstrated good precision across all analytes, with within-run and total coefficients of variation below 6.8%. Deming regression demonstrated high quantitative (r > 0.98) and high qualitative agreement (Kappa > 0.93) with the reference platforms. The combined multi-marker panel partially compensated for the low sensitivity of individual markers (e.g., SCC, 4.0%), improving overall diagnostic performance to a moderate level (AUC = 0.731). Statistical comparison indicated that the combined AUC of the evaluated system was lower than that of the Roche platform (0.895, P < 0.001) but comparable to the Abbott system (0.782, P > 0.05), while maintaining relatively high specificity (>90%). Conclusion Following preliminary verification, the system showed acceptable analytical performance and satisfactory reference concordance. While offering a cost-effective adjunctive tool for preliminary risk assessment, its clinical use requires external validation.
Purpose: Owing to the absence of appropriate pediatric reference intervals (RIs), this study aimed to establish age-specific RIs for serum biochemical analytes in healthy Chinese children. Methods: A total of 2627 residual serum samples were collected from children aged 28 days to 17 years at Shenzhen Baoan Women's and Children's Hospital. Forty-one analytes, including enzymes, proteins, lipids, electrolytes, and others, were measured using the Mindray BS-2800M analyzer. Outlier removal and RIs establishment were performed according to Clinical and Laboratory Standards Institute (CLSI) EP28 guideline. Age-dependent trajectories were visualized, and RIs were compared with multicenter studies. Additionally, the pediatric patterns of estimated glomerular filtration rate (eGFR) were also evaluated by equations. Results: Twenty-one analytes showed distinct age-dependent dynamics, including liver enzymes, prealbumin, bilirubin, immunoglobulins, renal biomarkers, cardiac enzymes, ferritin, and antistreptolysin O. Meanwhile, electrolytes, proteins, glucose, and other anemia markers remained stable across ages. RIs for thirty-six analytes were largely consistent with comparative studies, although quantitative discrepancies were noted for triglyceride and anti-streptolysin O. In 806 children, eGFR increased rapidly during the first two years of life, with inter-formula variability. The European Kidney Function Consortium equation produced distribution pattern closest to measured ranges from the comparative study. Conclusions: This study established comprehensive pediatric biochemical RIs for forty-one serum analytes, with twenty-one exhibiting distinct age-dependent dynamics. Comparisons with landmark studies showed good agreement for thirty-six analytes. The eGFR equation selection requires caution, particularly in young children. These RIs provide reliable evidence to support cross-regional harmonization and help reduce inappropriate use of references in children.
Background:Clinical mass spectrometry is recognized for its high specificity and sensitivity in quantifying small-molecule biomarkers in serum. However, its broad adoption in clinical settings has been limited by challenges such as low automation and time-consuming workflows. Methods:This study aimed to develop a rapid, sensitive, and automated magnetic solid-phase extraction(MSPE) method using Hydrophilic-Lipophilic Balance (HLB) magnetic bead-based sample preparation coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous quantification of homocysteine(Hcy) and its related nine metabolites. The method was comprehensively validated for specificity, linearity, sensitivity, accuracy, precision, matrix effects, carry-over; and compared it with solid-phase extraction (SPE) methods. Results:Results showed excellent linearity (r2 > 0.995) for all nine biomarkers associated with the homocysteine metabolic cycle. Both the limits of detection and quantification met the clinical requirements. Recoveries at low, medium, and high spiking levels ranged from 85.46% to 114.48%. Intra-day precision (CV) was between 0.82% and 7.63%, and inter-day precision (CV) ranged from 1.62% to 11.43%. Matrix effects were acceptable, with internal standard-normalized matrix factors ranging from 0.83 to 1.19. Carry-over rates were between -7.43% and 5.21%. Method comparison with protein precipitation sample preparation showed correlation coefficients from 0.9462 to 0.9957, indicating no systematic bias. Conclusions:In conclusion, the developed semi-automatization method is rapid, highly sensitive, and reproducible, making it suitable for quantitative analysis of these nine homocysteine cycle-related biomarkers in clinical serum samples. It provides a reliable analytical tool for the diagnosis, treatment, and prevention of associated diseases.
Background: Proper pre-analytical handling of blood samples is essential for reliable glucose measurement and for the accurate diagnosis and monitoring of diabetes mellitus. This study aimed to compare glucose concentrations in serum, lithium heparin (Li-Heparin), sodium fluoride citrate (NaF-Citrate), and sodium fluoride (NaF) plasma. Methods: Blood samples from 45 healthy volunteers were collected in Li-Heparin, NaF-Citrate, and serum tubes. Serum was stored at room temperature (RT) for up to 30 min, Li-Heparin whole blood in an ice-water slurry, and NaF-Citrate and Li-Heparin whole blood at RT for up to 10 min. Glucose was measured using the Cobas 8000 c701 analyzer. Additionally, internal routine laboratory data (n > 85,000) were analysed following the switch from NaF to NaF-Citrate tubes in 2015. Results: NaF-Citrate plasma showed the highest positive deviations, up to 3.11% compared with both reference conditions. Li-Heparin plasma at RT was 1.17 % higher than Li-Heparin plasma on ice. Serum glucose did not differ significantly from Li-Heparin plasma on ice. In routine data, the introduction of NaF-Citrate tubes resulted in an average increase of 10.5% in measured glucose concentrations in 2016. Conclusion: Glucose concentrations measured in NaF-Citrate plasma and Li-Heparin plasma (ice or RT) within 30 min after blood collection are not comparable. According to ADA and WHO diagnostic criteria, switching to NaF-Citrate tubes increased the proportion of patients with impaired fasting glucose by 16.9% and impaired glucose tolerance by 8.8%. NaF-Citrate tubes are recommended when immediate centrifugation and plasma separation cannot be ensured.
In clinical practice, assessing alcohol intake can be challenging because patients may under-report or misjudge their consumption, so biomarkers of chronic intake provide valuable additional information. Phosphatidylethanol (PEth) is a group of abnormal phospholipids that are formed in cell membranes only in the presence of ethanol (alcohol). We developed, analytically validated, and clinically implemented a rapid and robust LC-MS/MS-based method for quantifying PEth (16:0/18:1) in whole blood. The method demonstrates a limit of detection (LOD) of 0.01 μM and a limit of quantification (LOQ) of 0.03 μM, with confirmed linearity up to at least 0.8 μM. Validation using reference materials and samples from patients with liver disease and self-reported alcohol intake confirmed its accuracy. Clinical implementation over a two-month period, involving 20 analytical batches, demonstrated inter-assay precision below 15%. Our results support the clinical applicability of PEth use for accurate estimation of alcohol intake in humans.
Background Point-of-care testing (POCT) can accelerate clinical decision-making, but its analytical performance must be verified against central laboratory platforms before implementation. This study compared an immunofluorescence-based POCT system (STANDARD™ F Analyzer, SD Biosensor) with central laboratory analyzers used in the Clinical Laboratory of IRCCS MultiMedica (Siemens Atellica IM and CS-5100) for cardiac Troponin I (cTnI), creatine kinase-MB (CK-MB), N-terminal pro-brain natriuretic peptide (NT-proBNP), D-dimer and high-sensitivity C-Reactive Protein (hs-CRP). Methods Serum and citrate samples were tested in parallel on both systems. Analytical precision was assessed using a CLSI EP15-A3 5×3 design. Method comparison included Passing–Bablok regression, Bland–Altman analysis and categorical agreement assessed by Cohen’s κ at clinically relevant decision thresholds. Results A total of 217 samples were analyzed. The POCT system met predefined precision acceptance criteria for most biomarkers. Significant inter-method differences were particularly evident for CK-MB and D-dimer, whereas NT-proBNP showed smaller analytical discrepancies. hs-CRP showed acceptable categorical agreement despite limited precision performance at one control level. For cTnI, quantitative comparison at very low concentrations was limited by the higher limit of quantification of the POCT assay. Overall, categorical agreement at clinical decision thresholds ranged from moderate to high across biomarkers. Conclusion The STANDARD™ F Analyzer showed variable analytical performance, with limited analytical comparability to reference methods, particularly for hs-CRP and at low cTnI concentrations. Categorical agreement was moderate overall, but clinically relevant misclassifications were observed. Therefore, the POCT platform may serve as a complementary tool in settings without immediate access to a core laboratory, provided that its analytical limitations are recognized and results are interpreted using assay-specific thresholds.
Detecting donor DNA in blood samples after multiple allogeneic transfusions remains a persistent concern in forensic human identification, given the limited and variable data. Although some studies have reported no secondary DNA profiles, transfusion-associated microchimerism (TA-MC) has been observed in deceased individuals who received perimortem blood transfusions.In this case report, we present autosomal Short Tandem Repeat (aSTR) findings for a deceased male who received four units of plasma-reduced RBCs within 24 hours and 25 units over a period of eight months before his death. Given this extensive blood transfusion history, there was concern that donor DNA might persist and interfere with obtaining a reliable aSTR profile for paternity testing. To address this, we analyzed buccal swabs, skin, and muscle tissue, in addition to blood samples. No secondary aSTR profile was detected in the blood, and all four sample types produced concordant DNA profiles.In this instance, results suggest that any donor DNA present was below the detection threshold for STR analysis using capillary electrophoresis (CE) but may be detectable with more sensitive genotyping assays such as next generation sequencing (NGS) or digital PCR. Detection is more likely after a recent massive pRBC transfusion, the use of non-leukoreduced blood, or transfusion in leukopenic recipients. Despite the complications associated with massive allogeneic blood transfusions, aSTR typing remains a robust and reliable method for forensic human identification, particularly when suitable sample types are selected and rigorous procedures are followed in cases involving transfused individuals.
Objectives:Pre-analytical conditions are critical to ensure the reliability of laboratory results, as emphasized by ISO 15189 standards. Drone transport has emerged as a promising alternative to conventional logistics, but its impact on sample integrity remains insufficiently characterized. This pilot study aimed to assess the pre-analytical stability of blood samples transported by drone versus ground transport. Methods:In this prospective study, 30 healthy volunteers were included. Six blood tubes per participant were collected simultaneously and assigned to ground or drone transport (20 min). A panel of 23 biochemical, hematological, and hemostatic parameters was analyzed. Agreement between transport modalities was assessed using paired comparisons, coefficients of variation, intraclass correlation coefficients (ICC), Pearson correlation, and Bland-Altman analysis. Results:No clinically meaningful differences were observed between transport modalities. Mean values and variability were comparable across parameters. Most analytes showed excellent agreement, with ICC and Pearson correlation coefficients >0.90. Although ALT and LDH showed statistically significant differences (p = 0.039), these were small and clinically negligible. Bland-Altman analysis confirmed minimal bias for ALT (-0.63 U/L), whereas LDH exhibited wider limits of agreement, suggesting increased sensitivity to transport-related factors (-9.7 U/L). No hemolysis, temperature deviation, or safety incidents were observed. Conclusions:Drone transport ensures robust pre-analytical stability of blood samples across a wide range of laboratory parameters. However, analyte-specific variability, particularly for LDH, highlights the need for targeted validation. These findings support the integration of drone-based logistics into laboratory workflows, while emphasizing the importance of analyte-dependent evaluation.
