Pre-analytical Processing Research Articles

A-252 Evaluation of the Use of One-tube Blood Collection and Automated Specimen Processing System for LIAISON® QuantiFERON®-TB Gold Plus Testing at a Large Regional Reference Laboratory

Abstract Background LIAISON® QuantiFERON®-TB Gold Plus (QFT-Plus) is an in vitro diagnostic assay for the indirect detection of infection caused by Mycobacterium tuberculosis complex (MTBC). QFT-Plus testing consists of four tubes (controls and TB antigens) and detects interferon-γ (IFN-γ) released by host cells when exposed to MTBC peptide antigens using chemiluminescence immunoassay (CIA) technology. Patient’s blood can be collected directly into four QFT-Plus tubes or into a single lithium heparin tube. If the one-tube collection process is utilized, blood must be aliquoted into each of the four QFT-Plus tubes prior to testing. In this study, we evaluated the workflow and feasibility of lithium-heparin blood collection and pre-analytical processing using the Copan Universe system for QFT-Plus testing on the LIAISON XL analyzer. Methods Whole blood collected into a 6 mL lithium heparin tube was mixed using a tube rotator prior to processing on the Copan UniVerse, which aliquots 0.9 mL blood into each of four QFT-Plus tubes and label the tubes with patient information. The lithium heparin tube is mixed again immediately prior to aliquoting into each QFT-Plus tube. Different methods of mixing (vortexing vs pipetting) were assessed for efficiency and performance. Walkaway feasibility was also assessed by testing aliquoting efficiency when loading 6, 9, 12 and 18 blood samples at a time. Visual inspection of the QFT-Plus tubes to check accuracy of labels was performed prior to incubation and validation of readability of the barcode labels through the analyzer was performed. Results Of 44 blood samples that were pipet mixed, 26 (59.1%), 15 (34.1%) and 3 (6.8%) showed no, slight and moderate hemolysis, respectively. None were grossly hemolyzed. Vortex mixing showed similar results with 61.3% (19/31) and 38.7% (12/31) showed no or slight hemolysis. However, bubble formation, which may interfere with pipetting by the UniVerse, was observed in several tubes when the samples were vortex mixed. The UniVerse appropriately aliquoted 0.9 mL blood and labeled 280 tubes. Only 1 (0.4%) tube had <0.9 mL volume detected. It takes about 20 minutes to aliquot 12 patient samples into four QFT-Plus tubes. Walkaway was possible when loading 18 patients’ samples at a time without negative downstream impact on pipetting or appropriate mixing of samples for QFT-Plus testing. All QFT-Plus tubes were labeled accurately with barcodes and accession numbers which were all read and processed through the analyzer. Conclusions Implementation of lithium-heparin blood collection for centralized QFT-Plus testing was feasible when used in conjunction with the Copan UniVerse, which allowed for high throughput specimen processing and mitigated the risk of sample mislabeling.

A-182 Applications of artificial intelligence optical character recognition in laboratory clerical functions offsetting staffing shortages and error reduction

Abstract Background The clinical laboratory profession has been dealing with a decade long, and now critical retention and staffing problem. The average national turnover rate for laboratory technicians is 17%, and the highest turnover rate exhibited for phlebotomists is 36%. While turnover rates have an impact on a laboratory, or hospitals budget, more importantly they have a substantial impact on quality and error rates, particularly in the pre-analytical stage of testing; with studies showing up to 65.09% of errors impacting the pre-analytical phase. Methods In an effort to remedy this high error rate, a meta-analysis was performed on paper order entry and clerical pre-analytical error, in parallel with performance and accuracy data of advanced optical character recognition driven by artificial intelligence and machine learning (AI OCR). Results AI OCR studies demonstrated an untrained accuracy percentage, to include handwriting of 91.08%, and when trained utilizing both relational and hierarchal logic data bases accuracy increased to 98.4%. The rate at which AI OCR processed and classified medical information was 3-4 pages per second. It is estimated that 99.9%-100% accuracy can be achieved with additional form field training and mapping. The level of accuracy and efficiency for laboratory paper order-entry utilizing AI OCR can also be increased with the addition of natural language processing and data recognition algorithms when combined with logic databases containing information such as medical record numbers, ICD-10 codes, and laboratory test compendiums. Conclusions The performance of this technology demonstrates a strong application to reduce the burden of clerical paper order entry, while addressing the 30.8% dissatisfaction rate regarding manual entry processes among lab professionals surveyed. Moreover, AI OCR clerical data classification, extraction and entry processes augmented with human verification steps examined, demonstrated overall accuracy of 99.99%. The application of AI OCR in laboratory pre-analytical processes has the potential to improve test order accuracy and efficiency, ultimately impacting the quality of patient care and operational costs for healthcare facilities. When examined through the lens of staff retention, studies show that 80% of laboratory employees feel that a workplace with the necessary tools and technology positively impacts their job satisfaction. While organizations investing in technologies that drive both job efficiency and satisfaction see a 125% return on investment through productivity. Demonstrating that the application of sophisticated technologies that both increase productivity and decrease error rates have a substantial impact on job satisfaction and ultimately retention. Although laboratory clerical data and order entry processes were examined for this meta-analysis, the aggregate data compiled showed that the application of AI OCR in clerical processes could also have a substantial impact on the 25.9% annual turnover rate for hospitals as a whole in the United States.

A-060 Assessing Glucose Stability: A Comparison of Citrate/NaF, NaF, and Gel Separator Vacuum Tubes in Routine Medical Laboratory Practice

Abstract Background In cases where a glycolysis inhibitor is not utilized, blood cells in vitro can consume glucose. Although current guidelines suggest using citrate-buffered inhibitors or ice-water slurry to mitigate this, practical and financial constraints often hinder adherence to these recommendations. Our objective was to evaluate the difference in glucose concentration in our routine sample management by employing Citrate buffered/NaF, NaF, and Gel Barrier tubes. Methods In this study, we utilized Greiner FCmix citrate-buffered NaF and EDTA tubes (3 mL), Becton Dickinson NaF and EDTA tubes (4 mL), and Becton Dickinson SSTII Gel Barrier tubes (5 mL). Following venipuncture, the tubes were transferred to the laboratory and centrifuged within 30 minutes. Glucose levels of 46 patients' samples were measured using a Cobas 6000 system (c501) employing the Glucose Hexokinase method, which exhibits an intermediate coefficient of variation (CV) of approximately 1.5% and no significant bias, as confirmed by monitoring with Biorad EQAS. Results The mean plasma glucose difference between Citrate-buffered tubes and NaF tube samples was 2.8 mg/dL with a 95% confidence interval (CI) ranging from 2.2 to 3.4 mg/dL (Mean: 0.16 mmol/L; CI: 0.12 - 0.19 mmol/L). The difference in paired samples of NaF plasma and Gel Barrier tubes serum glucose was 3.2 mg/dL with a CI of 2.5 - 3.9 mg/dL (Mean: 0.18 mmol/L; CI: 0.14 - 0.22 mmol/L). Lastly, the mean difference in Citrate-buffered plasma and serum Gel barrier tubes for 46 paired samples was 6.0 mg/dL with a CI of 5.2 - 6.7 mg/dL (Mean: 0.33 mmol/L; CI: 0.29 - 0.37 mmol/L). Conclusions Following guidelines, Citrate-buffered/NaF tubes demonstrate efficacy as glucose stabilizers. When compared to NaF tubes alone within a 30-minute timeframe, the mean difference between the two types of tubes was 2.8 mg/dL. Although NaF tubes are no longer recommended for glucose testing in plasma, they still outperform gel barrier serum tubes by 3.2 mg/dL. Furthermore, compared to citrate/NaF Plasma, serum glucose levels were on average 6.0 mg/dL lower. The preanalytical sample stage poses a significant obstacle to its reliable use for diagnostic or monitoring purposes. Laboratories should consider transitioning to Citrate/NaF tubes, or employ mathematical corrections if utilizing gel separator tubes for glucose testing in their established preanalytical process.

MIBLood-EV: An Online Reporting Tool to Facilitate the Standardized Reporting of Preanalytical Variables and Quality Control of Plasma and Serum to Enhance Rigor and Reproducibility in Liquid Biopsy Research.

Pre-analytical variability significantly impacts the reproducibility of liquid biopsy research, which is critical for precision medicine and biomedical research. This report highlights the challenges and variability in the pre-analytical processes of liquid biopsies, especially regarding extracellular vesicles (EVs), which are crucial for diagnostics in oncology. The MIBlood-EV initiative aims to standardize the reporting of pre-analytical variables and the quality control of plasma and serum samples to enhance reproducibility in EV research. By providing a comprehensive and flexible reporting framework, MIBlood-EV seeks to improve the reliability of EV studies and facilitate the development of evidence-based protocols, ultimately advancing the field of liquid biopsy research.

Strategies for Error Reduction: Why More Stringent Premarket Evaluations Do Little to Prevent Laboratory Errors and Traffic Accidents

Laboratory testing is a complex process with a significant error rate. Studies of laboratory errors have found that the major causes are preanalytical factors, interferences, and process errors. Efforts by regulatory agencies to improve quality via more stringent premarket evaluations of laboratory tests therefore have poor prospects of reducing laboratory errors and improving test quality. Efforts toward increasing the regulation of laboratory tests are analogous to preventing traffic accidents by increasing the premarket evaluation of automobiles. This analogy illustrates how increased premarket evaluation has limited prospects for quality improvement and, in some cases, actually contributes to errors and lower quality. Tools that are used by laboratories to detect, prevent, and address analytical errors are discussed, and the increased implementation of such tools offers approaches that can be used to improve laboratory quality.

Correlation between serum advanced glycation end-products and vascular complications in patient with type 2 diabetes

Advanced glycation end-products (AGEs) formation increases with metabolic disorders, leading to higher serum AGE levels in patients with progressive vascular complications. Measuring AGE levels in biological samples requires multiple pre-analytical processing steps, rendering analysis of multiple samples challenging. This study evaluated the progression of diabetic complications by analyzing AGE levels using a pre-analytical processing strategy based on a fully automated solid phase-extraction system. Serum samples from patients with diabetes, with or without macrovascular complications (Mac or non-Mac) or microvascular complications (Mic or non-Mic), were processed with the established methods. Free and total AGE levels in sera were measured using liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). In patients with diabetes, both free and total AGE levels were elevated in those with complications compared to those without complications. In Mac and Mic groups, free and total AGE levels and z-scores (the sum of normalized AGE levels) also increased. AGE z-scores were markedly higher than those of single AGE levels in distinguishing each complication. Our study demonstrated that the free AGE z-score, measured using a new analytical method without hydrolysis, correlated with the presence of vascular complications and may serve as a marker of disease complications.

PrecivityAD2™ Blood Test: Analytical Validation of an LC-MS/MS Assay for Quantifying Plasma Phospho-tau217 and Non-Phospho-tau217 Peptide Concentrations That Are Used with Plasma Amyloid-β42/40 in a Multianalyte Assay with Algorithmic Analysis for Detecting Brain Amyloid Pathology.

Alzheimer's disease (AD) is a progressive irreversible neurodegenerative disorder that represents a major global public health concern. Traditionally, AD is diagnosed using cerebrospinal fluid biomarker analysis or brain imaging modalities. Recently, less burdensome, more widely available blood biomarker (BBM) assays for amyloid-beta (Aβ42/40) and phosphorylated-tau concentrations have been found to accurately identify the presence/absence of brain amyloid plaques and tau tangles and have helped to streamline AD diagnosis. However, few BBMs have been rigorously analytically validated. Herein, we report the analytical validation of a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) multiplex method for quantifying plasma phosphorylated-tau217 (p-tau217) and non-phosphorylated-tau217 (np-tau217) peptide concentrations. We combined the p-tau217/np-tau217 concentrations ratio (%p-tau217) and the previously validated LC-MS/MS multiplex assay for plasma Aβ42/40 into a new multianalyte assay with algorithmic analysis (MAAA; PrecivityAD2™ test) that identifies brain amyloid status based on brain amyloid positron emission tomography. We found (a) the %p-tau217 assay is precise, accurate, sensitive, and linear over a wide analytical measurement range, and free from carryover and interference; (b) the pre-analytical specimen collection, processing, storage, and shipping conditions that maintain plasma tau peptide stability; and (c) using the measured analytical imprecision for plasma Aβ42/40 and p-tau217/np-tau217 levels in a worst-case scenario model, the PrecivityAD2 test algorithm for amyloid pathology classification changed for only 3.5% of participants from brain amyloid positive to negative, or from negative to positive. The plasma sample preparation and LC-MS/MS methods underlying the PrecivityAD2 test are suitable for use in the clinical laboratory and valid for the test's intended purpose: to aid in the diagnostic evaluation of individuals aged 55 and older with signs or symptoms of mild cognitive impairment or dementia.

Engineering of a DNA/γPNA Hybrid Nanoreporter for ctDNA Mutation Detection via γPNA Urinalysis.

Detection of circulating tumor DNA (ctDNA) mutations, which are molecular biomarkers present in bodily fluids of cancer patients, can be applied for tumor diagnosis and prognosis monitoring. However, current profiling of ctDNA mutations relies primarily on polymerase chain reaction (PCR) and DNA sequencing and these techniques require preanalytical processing of blood samples, which are time-consuming, expensive, and tedious procedures that increase the risk of sample contamination. To overcome these limitations, here the engineering of a DNA/γPNA (gamma peptide nucleic acid)hybrid nanoreporter is disclosed for ctDNA biosensing via in situ profiling and recording of tumor-specific DNA mutations. The low tolerance of γPNA to single mismatch in base pairing with DNA allows highly selective recognition and recording of ctDNA mutations in peripheral blood. Owing to their remarkable biostability, the detached γPNA strands triggered by mutant ctDNA will be enriched in kidneys and cleared into urine for urinalysis. It is demonstrated that the nanoreporter has high specificity for ctDNA mutation in peripheral blood, and urinalysis of cleared γPNA can provide valuable information for tumor progression and prognosis evaluation. This work demonstrates the potential of the nanoreporter for urinary monitoring of tumor and patient prognosis through in situ biosensing of ctDNA mutations.

Advances in Blood Biomarkers for Alzheimer's Disease: Ultra-Sensitive Detection Technologies and Impact on Clinical Diagnosis.

Alzheimer's disease has escalated into a critical public health concern, marked by its neurodegenerative nature that progressively diminishes cognitive abilities. Recognized as a continuously advancing and presently incurable condition, AD underscores the necessity for early-stage diagnosis and interventions aimed at delaying the decline in mental function. Despite the proven efficacy of cerebrospinal fluid and positron emission tomography in diagnosing AD, their broader utility is constrained by significant costs and the invasive nature of these procedures. Consequently, the innovation of blood biomarkers such as Amyloid-beta, phosphorylated-tau, total-tau et al, distinguished by their high sensitivity, minimal invasiveness, accessibility, and cost-efficiency, emerges as a promising avenue for AD diagnosis. The advent of ultra-sensitive detection methodologies, including single-molecule enzyme-linked immunosorbent assay and immunoprecipitation-mass spectrometry, has revolutionized the detection of AD plasma biomarkers, supplanting previous low-sensitivity techniques. This rapid advancement in detection technology facilitates the more accurate quantification of pathological brain proteins and AD-associated biomarkers in the bloodstream. This manuscript meticulously reviews the landscape of current research on immunological markers for AD, anchored in the National Institute on Aging-Alzheimer's Association AT(N) research framework. It highlights a selection of forefront ultra-sensitive detection technologies now integral to assessing AD blood immunological markers. Additionally, this review examines the crucial pre-analytical processing steps for AD blood samples that significantly impact research outcomes and addresses the practical challenges faced during clinical testing. These discussions are crucial for enhancing our comprehension and refining the diagnostic precision of AD using blood-based biomarkers. The review aims to shed light on potential avenues for innovation and improvement in the techniques employed for detecting and investigating AD, thereby contributing to the broader field of neurodegenerative disease research.

Design of metabolomic studies: pre-analytical process

Metabolomics is the comprehensive analysis of small molecules involved in metabolic pathways that control biochemical processes and functions of cells in the body. Metabolomic studies consist of three major steps: pre-analytical, analytical and post-analytical. The review emphasizes the importance of the pre-analytical stage, which is the journey of a biosample from the patient to the biobank and then to the analytical laboratory. Based on the literature analysis, the review presents the factors that influence the quality of the sample and therefore the quality of the final research result: clinical information collection, biosample selection, biosample collection and processing, and subsequent storage. Proper design of metabolomic studies, quality control of samples from collection to analysis by physicochemical methods provides data that can improve the quality of disease diagnosis, provide a transition to personalized medicine.

A novel molecular assay conducted on the BD Max system to facilitate reflex testing for vanA and vanB in clinical isolates of enterococci

Infections caused by vancomycin-resistant enterococci (VRE) are common. Real-time PCR assays targeting vanA and vanB facilitate screening of patients in healthcare settings to limit the risk of dissemination, especially amongst those at high-risk of infection or with limited treatment options. Such assays are commonly performed as reflex testing procedures where they augment phenotypic techniques and shorten turnaround time to benefit timely clinical management. 'Random access' and 'sample-to-result' real-time PCR platforms are suited for this application as they are of low complexity and less technically demanding. Modelled on these attributes, we configured a real-time PCR assay (VRE BD) for detection of vanA/B in clinical isolates of enterococci, adapted for the BD Max System (Becton Dickinson). We applied an unconventional approach by testing suspensions of microorganisms in water to circumvent the traditional pre-analytical genomic extraction process. Our objective of this study was to assess the performance of this assay for detection of VRE in cultures by validating against a traditional real-time PCR assay based on the LightCycler 2.0 platform (Roche, VRE RO). A high level of analytical sensitivity and specificity (≥99.0%) for both genes was obtained when testing suspensions derived from blood agar. Results for suspensions obtained from chromID VRE (Edwards Group) showed a similar level of performance for vanA detection (100%), but not for the vanB target (≥90.9%) where a lesser number of isolates were available for testing. However, our results for VRE detection in isolates from these media were repeatable and reproducible, and equated to positive and negative predictive values of ≥95.2% and ≥97.8%, respectively. Furthermore, the VRE BD assay was also able to accurately detect VRE in clinical and spiked BacT/ALERT (bioMérieux) blood cultures. Thus, the technical simplicity, short turnaround time and robustness of this high performing assay for VRE is suitable for reflex testing. In addition, the format developed for the BD Max platform has potential application for reflex testing other molecular targets of clinical importance.

Evaluation of Blood Transfusion Request form: The Experience in a Tertiary Health Facility in Jos, Nigeria

Background: There is a thin line between a safe blood transfusion and transfusion-related fatality hence the need to be diligent in every aspect of the blood transfusion process. Appropriate and complete documentation on a blood transfusion request form is one of the most important preanalytic activities serving as a communication tool between the clinician and the blood transfusion laboratory personnel. Aims: To evaluate compliance with appropriate and complete documentation of information on our blood transfusion request forms for a reliable preanalytic process towards an efficient blood transfusion service. Study Design: It is a retrospective study. Place and Duration of Study: Blood Bank of the Jos University Teaching Hospital between January to December 2023. Methodology: Six thousand, three hundred and sixty (6360) blood transfusion request forms from the Jos University Teaching Hospital Blood bank were evaluated for complete or incomplete documentation retrospectively and results were presented in frequencies and percentages. Results: There was 100% compliance in filling in the patients' surnames and other names as well as the laboratory number and blood groups of the patients while only 4779 (75.14%) filled in the patients' ages with 1416(22.26%) using the prefix of adult(ad) while 165(2.59%) fail to document the patients' age. There were 2829 (44.48%) males with 3522 (55.38%) females while no sex was indicated in 9 of the reviewed forms. Obstetrics history has the least cumulated documented response of 0.38% while a significant 1008 (15.85%) did not indicate either blood grouping or blood grouping with cross-match request. Conclusion: Appropriate and complete documentation of information on blood transfusion request forms is a problem among clinicians and will require continuous education on its importance, periodic auditing, provision of electronic data system and attitudinal change for a better blood transfusion compatibility service.

Hyaluronic Acid and Large Extracellular Vesicles (EVs) in Synovial Fluid and Plasma of Patients With End-Stage Arthritis: Positive Association of EVs to Joint Pain.

Hyaluronic acid (HA) in synovial fluid (SF) contributes to boundary lubrication with altered levels in osteoarthritis (OA) and rheumatoid arthritis (RA). SF extracellular vesicles (EVs) may participate in arthritis by affecting inflammation and cartilage degradation. It remains unknown whether HA and EVs display joint-specific alterations in arthritic SFs. We investigated the numbers and characteristics of HA-particles and large EVs in SF from knees and shoulders of 8 OA and 8 RA patients and 8 trauma controls, and in plasma from 10 healthy controls and 11 knee OA patients. The plasma and SF HA concentrations were determined with a sandwich-type enzyme-linked sorbent assay, and EVs and HA-particles were characterized from plasma and unprocessed and centrifuged SFs with confocal microscopy. The data were compared according to diagnosis, location, and preanalytical processing. The main findings were: (1) OA and RA SFs can be distinguished from trauma joints based on the distinctive profiles of HA-particles and large EVs, (2) there are differences in the SF HA and EV characteristics between shoulder and knee joints that could reflect their dissimilar mobility, weight-bearing, and shock absorption properties, (3) EV counts in SF and plasma can positively associate with pain parameters independent of age and body adiposity, and (4) low-speed centrifugation causes alterations in the features of HA-particles and EVs, complicating their examination in the original state. Arthritis and anatomical location can affect the characteristics of HA-particles and large EVs that may have potential as biomarkers and effectors in joint degradation and pain.

Pre-analytical challenges from adsorptive losses associated with thiamine analysis

Thiamine (vitamin B1) is an essential vitamin serving in its diphosphate form as a cofactor for enzymes in the citric acid cycle and pentose-phosphate pathways. Its concentration reported in the pM and nM range in environmental and clinical analyses prompted our consideration of the components used in pre-analytical processing, including the selection of filters, filter apparatuses, and sample vials. The seemingly innocuous use of glass fiber filters, glass filter flasks, and glass vials, ubiquitous in laboratory analysis of clinical and environmental samples, led to marked thiamine losses. 19.3 nM thiamine was recovered from a 100 nM standard following storage in glass autosampler vials and only 1 nM of thiamine was obtained in the filtrate of a 100 nM thiamine stock passed through a borosilicate glass fiber filter. We further observed a significant shift towards phosphorylated derivatives of thiamine when an equimolar mixture of thiamine, thiamine monophosphate, and thiamine diphosphate was stored in glass (most notably non-silanized glass, where a reduction of 54% of the thiamine peak area was observed) versus polypropylene autosampler vials. The selective losses of thiamine could lead to errors in interpreting the distribution of phosphorylated species in samples. Further, some loss of phosphorylated thiamine derivatives selectively to amber glass vials was observed relative to other glass vials. Our results suggest the use of polymeric filters (including nylon and cellulose acetate) and storage container materials (including polycarbonate and polypropylene) for thiamine handling. Losses to cellulose nitrate and polyethersulfone filters were far less substantial than to glass fiber filters, but were still notable given the low concentrations expected in samples. Thiamine losses were negated when thiamine was stored diluted in trichloroacetic acid or as thiochrome formed in situ, both of which are common practices, but not ubiquitous, in thiamine sample preparation.

An end to end continuous quality improvement initiative to reduce errors in pre-analytical processes

An end to end continuous quality improvement initiative to reduce errors in pre-analytical processes

Control of non-conformities in the pre-analytical process in the clinical biology laboratory

Control of non-conformities in the pre-analytical process in the clinical biology laboratory

Assessment of Pre-analytical Errors and Fostering Strategies to Enhance Accurate Results and Efficient Turnaround Times in the Cytology Laboratory of a Tertiary Care Hospital.

Introduction Pre-analytical errors in cytology laboratories can significantly impact the accuracy of diagnostic results and turnaround times, ultimately affecting patient care. This article presents an evaluation of pre-analytical errors and proposes fostering strategies to enhance accuracy and efficiency in the cytology laboratory of a tertiary care hospital. The background discusses the importance of pre-analytical processes in ensuring reliable cytological diagnoses and the common errors encountered in specimen collection, handling, and transportation. Strategies for error reduction and improvement in turnaround times include staff education, standardization of procedures, utilization of appropriate collection and transport devices, implementation of quality control measures, and utilization of automation technologies. By addressing pre-analytical errors and implementing fostering strategies, cytology laboratories can optimize diagnostic accuracy, improve patient care outcomes, and enhance overall laboratory efficiency. Aims and objectives This study aims to assess the prevalence and nature of pre-analytical errors in the cytology laboratory of a tertiary care hospital to understand the extent of the issue, identify the specific factors contributing to pre-analytical errors like specimen collection, handling, and transportation processes, and pinpoint areas for improvement. It seeks to evaluate the impact of pre-analytical errors on the accuracy of cytological results and the efficiency of turnaround times, highlighting the consequences for patient care. Furthermore, the study aims to develop targeted strategies to minimize pre-analytical errors and enhance the accuracy of cytological results. Materials and methods This study was conducted at the Cytology Laboratory of our hospital from January 2023 to December 2023 after getting proper approval from the Institutional Review Board (IRB approval number 101/02/2024/PG/SRB/SMCH). It is a retrospective analytical study, and a total of 5412 samples from patients of the outpatient (OP) department, inpatient (IP) department, and community health outreach program facilities received in the cytology laboratory were analyzed during the study period. The inclusion criteria were the test samples sent specifically for cytological analysis. The samples sent for biochemical or microbiological examination were excluded. The frequency of sample distribution and rejected samples were calculated and the results were correlated. Results A total of 5,412 samples received in the cytology laboratory were analyzed during the study period. The majority of the samples were Papanicolaou smears (2,352, 43.5%), followed by fluid cytology (1,008, 18.6%) and ultrasound-guided fine-needle aspiration cytology (FNAC, 984, 18.2%). Of the total number of samples, 225 (4.16%) were repeated and 27 (0.5%) were rejected. Conclusions Pre-analytical, analytical, and post-analytical processes are the three key factors that determine the dependability and precision of cytological test results. Detecting critical alerts such as the positivity of malignancy underscores the paramount importance of result accuracy. Implementing good laboratory practices and conducting both external and internal audits can reduce the frequency of preventable errors in a cytology laboratory, thereby ensuring enhanced precision and expedited outputs.

Evaluation of cryoprotein investigation using a digital external quality assurance scheme.

Background: Robust preanalytical and analytical processes are critical for the detection of cryoproteins. There is significant variation in practice in the detection, analysis and reporting. Results: A survey in 2018 of 137 laboratories participating in the UK National External Quality Assessment Service (UK NEQAS) (6) quality control program showed significant variation in the laboratory processes which highlighted the need for standardisation of the detection, analysis and reporting of cryoglobulins.Conclusion: The first available EQA scheme aiming to harmonise practice for cryoprotein testing has been developed by UK NEQAS and laboratories should participate in an appropriate EQA scheme to fulfil requirements for ISO accreditation.

Pre-analytical stability of the CEA, CYFRA 21.1, NSE, CA125 and HE4 tumor markers.

For lung cancer, circulating tumor markers (TM) are available to guide clinical treatment decisions. To ensure adequate accuracy, pre-analytical instabilities need to be known and addressed in the pre-analytical laboratory protocols. This study investigates the pre-analytical stability of CA125, CEA, CYFRA 21.1, HE4 and NSE for the following pre-analytical variables and procedures; i) whole blood stability, ii) serum freeze-thaw cycles, iii) electric vibration mixing and iv) serum storage at different temperatures. Left-over patient samples were used and for every investigated variable six patient samples were used and analysed in duplicate. Acceptance criteria were based on analytical performance specifications based on biological variation and significant differences with baseline. Whole blood was stable for at least 6 hours for all TM except for NSE. Two freeze-thaw cycles were acceptable for all TM except CYFRA 21.1. Electric vibration mixing was allowed for all TM except for CYFRA 21.1. Serum stability at 4°C was 7 days for CEA, CA125, CYFRA 21.1 and HE4 and 4 hours for NSE. Critical pre-analytical processing step conditions were identified that, if not taken into account, will result in reporting of erroneous TM results.

Faster clinical decisions in B-cell acute lymphoblastic leukaemia: A single flow cytometric 12-colour tube improves diagnosis and minimal residual disease follow-up.

Assessing minimal residual disease (MRD) in B-cell precursor acute lymphoblastic leukaemia (BCP-ALL) is essential for adjusting therapeutic strategies and predicting relapse. Quantitative polymerase chain reaction (qPCR) is the gold standard for MRD. Alternatively, flow cytometry is a quicker and cost-effective method that typically uses leukaemia-associated immunophenotype (LAIP) or different-from-normal (DFN) approaches for MRD assessment. This study describes an optimized 12-colour flow cytometry antibody panel designed for BCP-ALL diagnosis and MRD monitoring in a single tube. This method robustly differentiated hematogones and BCP-ALL cells using two specific markers: CD43 and CD81. These and other markers (e.g. CD73, CD66c and CD49f) enhanced the specificity of BCP-ALL cell detection. This innovative approach, based on a dual DFN/LAIP strategy with a principal component analysis method, can be used for all patients and enables MRD analysis even in the absence of a diagnostic sample. The robustness of our method for MRD monitoring was confirmed by the strong correlation (r = 0.87) with the qPCR results. Moreover, it simplifies and accelerates the preanalytical process through the use of a stain/lysis/wash method within a single tube (<2 h). Our flow cytometry-based methodology improves the BCP-ALL diagnosis efficiency and MRD management, offering a complementary method with considerable benefits for clinical laboratories.