Serum Separator Tubes Research Articles

Differences Between Serum and Plasma: An Indirect Approach to Derive a Combined Reference Interval for Total Protein, Albumin, and Globulin

Abstract The objective of our study is to derive reference intervals (RI) for total protein (TP), albumin and globulin that are appropriate for both plasma and serum samples by applying statistical analyses to retrospective patient data. Both plasma and serum are acceptable matrices for these tests at our institution, with a single RI that applies to all specimens. The widespread shortage of lithium heparin (LH) tubes that occurred as a consequence of the pandemic prompted our lab to switch the primary tube type for comprehensive metabolic panels (CMPs) from LH to serum separator tubes (SST) in October 2021. The primary tube type reverted to LH in October 2023, and no adjustments were made to RIs at either time. To determine how well our RIs align with the accepted matrices, we used R Statistical Programing version 4.3.1 to apply an indirect, nonparametric statistical approach to data extracted from the laboratory information system (LIS) between July 2018 and December 2023. All orders for TP, albumin and globulin were queried to include the following components in the analysis: TP, albumin, globulin, serum TP, serum albumin and serum globulin. The records were divided into 3 epochs based on the primary tube type for CMP on the order date, with epochs 1 and 3 reflecting LH and epoch 2 reflecting SST. We attempted to identify a subset of serum and plasma samples in healthy individuals by applying exclusion criteria based on inpatient status, age, and draw station at which the specimen was collected. For each component, a resampling bootstrap method was used to calculate the RIs and corresponding 95% confidence intervals by epoch. Our results show that the RI of TP, albumin and globulin changed significantly between the epochs, reflecting the difference in primary matrix. The components serum TP, serum albumin, and serum globulin did not see a significant change in values over the epochs. A verification study looking at plasma and serum samples in 20 healthy individuals support these results. Based on these findings, our reference intervals were updated from 6.6-8.7 g/dL to 5.9-8.3 g/dL for TP, from 3.6-4.9 g/dL to 3.6-5.1 g/dL for albumin and from 2.3-3.5 g/dL to 1.5-3.9 g/dL for globulin. We show that there are statistically and potentially clinically significant differences in the reference intervals of TP, albumin and globulin in serum compared to plasma. This study demonstrates a big data analytics approach to derivation of RIs and emphasizes the importance of RI assessment and adjustment for patient population and acceptable specimen type.

A-070 Evaluation of Plain and Serum-Separator Blood Collection Tubes for the Measurement of Steroid Hormones by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Abstract Background Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) is the preferred method for quantifying steroids. Commonly measured steroids 11-deoxycortisol, androstenedione, testosterone, 17-hydroxyprogesterone and dehydroepiandrosterone (DHEA). Although serum separator blood collection tubes may not be recommended but their frequent use is attributed to the ease of serum separation. Moreover, the serum separator tube may be the only available option for analysis, without the need for additional sample collection. Under such conditions, from certain serum separator tubes, we observed occasional interference in LC-MS/MS steroid method. We pursued to evaluate the performance of blood collection tubes without and those with serum separators, for the quantification of specified steroids by LC-MS/MS. Methods Blood samples were collected from 10 volunteers using both plain and serum separator blood collection tubes procured from two major suppliers (Greiner and BD). The specified steroids were measured by LC-MS/MS method. The procedure involved addition of labeled internal standards to the samples and extraction of steroids in methyl tert-butyl ether. Subsequently, the extract dried under a stream of nitrogen, and the residue was reconstituted in the mobile phase. LC-MS/MS analysis involved multiple reaction monitoring (MRM) for the specified steroids and their internal standards. Results The figure shows a sample chromatogram from a subject with blood samples collected in a plain (A) and serum separator tubes from Greiner (B) and BD (C1 and C2, a zoomed of C1). In this sample, from the plain tube, the concentrations of 11-deoxycortisol, androstenedione, testosterone, 17-hydroxyprogesterone and DHEA were 27, 55, 690, 99 and 283 ng/mL respectively. No significant interference was observed in Greiner serum separator tube. Whereas BD serum separator tube exhibited several erroneous peaks making the quantitation of several steroids challenging or inaccurate. Conclusions In the described LC-MS/MS method, BD serum-separator tubes exhibited erroneous peaks. These peaks were not seen in Greiner serum-separator tubes.

B-357 Siemens CA 72-4: an assay for measurement of Tumor Associated Glycoprotein (TAG) 72

Abstract Background The tumor associated glycoprotein (TAG) 72, also known as CA 72-4 is a mucin protein of high molecular weight. CA 72-4 is found on the surface of many cancer cells, including stomach (gastric), ovary, breast, colon and pancreatic cells. CA 72-4 is currently one of the primary markers for the diagnosis of gastric cancer and monitoring the course and efficacy of gastric cancer. In ovarian cancer, it has a higher sensitivity for mucin- type ovarian cancer, and its combination with CA125 for detection can significantly improve the clinical sensitivity. In colorectal cancer, CA 72-4 combined with CEA detection was also found to significantly improve the sensitivity of the initial diagnosis. An assay for CA 72-4 on Siemens Atellica IM (Siemens CA 72-4) is being developed and analytical performance is being presented. Methods The Siemens CA 72-4 assay (in development) is a one-step chemiluminescent microparticle immunoassay (CMIA) for the quantitative determination of CA 72-4 in human serum and plasma. The specimen is incubated with paramagnetic microparticles coated with the monoclonal antibody (mAb) B72.3 and acridinium-labeled mAb CC49 conjugate on the Siemens Atellica IM® System. After incubation and washing, Acid and Base reagents are added. The resulting relative light units (RLUs) are directly proportional to the amount of CA 72-4 in the sample allowing for the quantitative determination of CA 72-4 in the specimen. Results The following performance were determined using 3 unique lots of reagents and calibrators with the calibration range for the assay being 0.00 to 200.00 U/mL. The limit of blank (LoB), limit of detection (LoD), and limit of quantitation at 20% CV (LoQ) were determined and met the goal of ≤ 1.00 U/mL. Linearity according to CLSI EP06 Ed2 was demonstrated for a range of 1.00 through 200.00 U/mL. A Precision study of 2 controls and 5 panels spanning the range of the assay demonstrated a total %CV ≤ 6% for samples ≥ 5.00 U/mL and ≤ 0.25 SD for samples <5.00 U/mL. In the sample tube type study, a matrix comparison of 23 matched serum and plasma samples were evaluated. Passing-Bablock slopes of < 8% and r > 0.99 were observed when evaluating samples within the measurement range for plasma (LiHep, LiHep separator, K2EDTA and K3EDTA) tubes and serum separator tubes (SST) compared to the Red Top serum samples. Five endogenous substances were evaluated for interference in the CA 72-4 assay. The average percent difference between test and control samples for all endogenous interferents was < 10%. Percent difference in the presence of 28 potentially interfering drugs was ≤ 4.0%. Conclusions The Siemens CA 72-4 assay currently in development is a sensitive and precise assay for the quantitative determination of CA 72-4 in human serum and plasma.

B-356 Siemens CYFRA 21-1: an assay for measurement of cytokeratin fragment 21-1

Abstract Background Cytokeratins are structural proteins forming the subunits of epithelial intermediary filaments. The soluble fragment of cytokeratin 19, cytokeratin fragment 21-1 (CYFRA 21-1), can be found in the blood of lung cancer patients, primarily those with non-small cell lung cancer (NSCLC). CYFRA 21-1 is the preferred biomarker in the management of NSCLC, particularly squamous cell and large cell carcinoma subtypes. An assay for CYFRA 21-1 on Siemens ADVIA Centaur® XPT and Siemens Atellica® IM (Siemens CYFRA) is being developed and analytical performance is being presented. Methods The Siemens CYFRA 21-1 assay (in development) is a one-step chemiluminescent microparticle immunoassay (CMIA) for the quantitative determination of CYFRA 21-1 in human serum and plasma. The specimen is incubated with paramagnetic microparticles coated with the monoclonal antibody (mAb) BM19.21 and acridinium-labeled mAb KS19.1 conjugate on the Siemens ADVIA Centaur XPT System and Siemens Atellica IM System. After incubation and washing, Acid and Base reagents are added. The resulting relative light units (RLUs) are directly proportional to the amount of CYFRA 21-1 in the sample allowing for the quantitative determination of CYFRA 21-1 in the specimen. Results The following performance were determined using 3 unique lots of reagents and calibrators with the calibration range for the assay being 0.00 to 100.00 ng/mL. For the ADVIA Centaur XPT and Atellica IM systems were shown to be equivalent within 4%. The limit of blank (LoB), limit of detection (LoD), and limit of quantitation at 20% CV (LoQ) were determined and met the goal of ≤ 1.00 ng/mL. Linearity according to CLSI EP06 Ed2 was demonstrated for a range of 0.66 through 100.00 ng/mL. A Precision study of 2 controls and 5 panels spanning the range of the assay demonstrated a total %CV ≤ 8% at all levels. In the sample tube type study, a matrix comparison of 61 matched serum and plasma samples were evaluated. Passing-Bablock slopes of < 5% and r > 0.99 were observed when evaluating samples within the measurement range for plasma (LiHep, LiHep separator, K2EDTA and K3EDTA) tubes and serum separator tubes (SST) compared to the Red Top serum samples. Six endogenous substances were evaluated for interference in the CYFRA 21-1 assay. The average percent difference between test and control samples for all endogenous interferents was < 10%. Percent difference in the presence of 20 potentially interfering drugs was ≤ 4.0%. Conclusions The Siemens CYFRA 21-1 assay currently in development is a sensitive and precise assay for the quantitative determination of CYFRA 21-1 in human serum and plasma.

B-358 Siemens NSE: an assay for measurement of human Neuron-Specific Enolase

Abstract Background Neuron-specific enolase (NSE), a glycolytic enzyme (2-phospho-D-Glycerate hydrolase) can be found in a variety of non-neuroendocrine cells. Determination of NSE is used for monitoring response to therapy and detection of recurrent disease, such as SCLC and neuroendocrine tumors. An assay for NSE on Siemens ADVIA Centaur® XPT System and Siemens Atellica® IM system (Siemens NSE) is being developed and analytical performance is being presented. Methods The Siemens NSE assay (in development) is a one-step chemiluminescent microparticle immunoassay (CMIA) for the quantitative determination of NSE in human serum. The specimen is incubated with paramagnetic microparticles coated with the monoclonal antibody (mAb) NSE21, and acridinium-labeled mAb NSE17 conjugate on the Siemens ADVIA Centaur XPT system and Siemens Atellica IM system. After incubation and washing, Acid and Base reagents are added. The resulting relative light units (RLUs) are directly proportional to the amount of NSE in the sample allowing for the quantitative determination of NSE in serum. Results The following performance were determined using 3 unique lots of reagents and calibrators with the calibration range for the assay being 0.0 to 400.0 ng/mL. For the ADVIA Centaur XPT and Atellica IM systems were shown to be equivalent within 2%. The limit of blank (LoB), limit of detection (LoD), and limit of quantitation at 20% CV (LoQ) were determined and met the goal of ≤ 1.6 ng/mL. Linearity according to CLSI EP06 Ed2 was demonstrated for a range of 1.6 through 400.0 ng/mL. A Precision study of 2 controls and 6 panels spanning the range of the assay demonstrated a total %CV ≤ 7% at all levels. Comparison of 50 specimens collected in Red Top serum and Serum Separator Tubes (SST) had a Passing-Bablock slope of < 2% and r of > 0.99 when evaluating samples within the measurement range. Five endogenous substances were evaluated for interference in the Siemens NSE assay. The average percent difference between test and control samples for all endogenous interferents was < 10%. Percent difference in the presence of 38 potentially interfering drugs was ≤ 10.0%. The potential cross-reactant αα enolase was evaluated at concentration of 900 ng/mL and the percent cross-reactivity was ≤ 2%. Conclusions The Siemens NSE assay currently in development is a sensitive and precise assay for the quantitative determination of NSE in human serum.

HEMATOLOGY AND BIOCHEMICAL REFERENCE INTERVALS FOR FREE-RANGING PRONGHORN (ANTILOCAPRA AMERICANA) IN WEST TEXAS.

Pronghorn (Antilocapra americana) are considered a keystone species of North American grasslands and an important economic source for many landowners in Texas. Pronghorn restoration projects routinely capture and translocate individuals from surplus populations to restoration areas. The objective of this study was to generate normal hematological and biochemical reference intervals (RI) for free-ranging pronghorn populations in Texas as a health monitoring tool for pronghorn restoration efforts. Blood samples were collected by jugular venipuncture and divided among an EDTA tube, serum separator tube, and a single blood smear on site. Complete blood counts and biochemical profiles were completed at the Texas Veterinary Medical Diagnostic Laboratory. In total, 417 individuals (41 males, 376 females) were included in the analysis. RI were determined by robust methods (R Studio) and mixed models' analysis of variance (SPSS 28) to examine differences in blood parameters due to fever, sex, age (adult versus yearling [<1 yr of age]), cell abnormalities, and pathogen exposure reported by the testing laboratory. Sex, age, and pathogen exposure affected mean blood values, but did not warrant development of separate RI by class. Bluetongue virus was identified in 46.8% (195/417) of pronghorns and epizootic hemorrhagic disease in 89.4% (194/217) of pronghorns; 84.8% (184/217) of the pronghorns tested positive for both diseases. This information provides baseline hematology and biochemical parameters to assess the health of free-ranging pronghorn and guide wildlife managers in decision-making for future translocations and restoration objectives.

Assessing the stability of uncentrifuged serum and plasma analytes at various post-collection intervals

Abstract Objectives Our study aimed to assess the stability of 26 biochemistry analytes in serum or plasma samples separated from blood samples centrifuged at different time intervals after collection, simulating sample transport via despatch delivery systems. Methods Blood from forty-one volunteers were collected using five serum separator tubes (SST) and five fluoride oxalate tubes (FOT) for each volunteer following written informed consent. Each of the five tubes in both groups of SST and FOT was centrifuged at one of the time intervals: 0.5 h, 4 h, 8 h, 12 and 24 h after collection. These samples were left standing prior to centrifugation at room temperature. We calculated the percentage difference for each analyte between the 0.5 h and other time intervals to assess analyte stability. The percentage difference was compared to the desirable specification for bias and reference change value (RCV). Results Mean concentration of serum potassium showed a significant increase in the percentage RCV after 8 h, while CKMB showed an increase after 12 h of delayed centrifugation compared to the baseline (0.5 h). There were no significant percentage RCV for the other analytes at all timelines. Conclusions Serum potassium and CKMB were stable up to 8 and 12 h of delayed centrifugation respectively whilst all other analytes appear stable up to 24 h, suggesting that sample transport delay of up to 8 h, with the condition that room temperature is maintained, may not have a significant impact on accuracy of the biochemistry/immunochemistry test results.

Evaluation of Direct Antimicrobial Susceptibility Testing of Gram-Negative Bacilli and Staphylococcus aureus from Positive Pediatric Blood Culture Bottles Using BD Phoenix M50.

Bloodstream infections (BSIs) are life-threatening infections for which a timely initiation of appropriate antimicrobial therapy is critical. Antibiotic susceptibility testing (AST) directly performed on positive blood culture broths can help initiate targeted antibiotic therapy sooner than the standard AST performed on colonies isolated on solid media after overnight incubation. Faster antimicrobial susceptibility testing (AST) results can improve clinical outcomes, and reduce broad-spectrum antimicrobial consumption and healthcare-associated costs in sepsis. In this study, we evaluated the accuracy of a direct AST inoculation method on the BD Phoenix M50 system using serum separator tubes to harvest bacteria from positive pediatric blood culture bottles. Direct AST was performed on 132 monomicrobial pediatric blood culture bottles that were positive for Enterobacterales (65; 49.2%), Staphylococcus aureus (46; 34.8%), and non-fermenting Gram-negative bacilli (21; 16%). Overall, the categorical and essential agreements between the direct method and standard method were 99.6% and 99.8%, respectively. Very major, major, and minor error rates were 0.1%, 0.09%, and 0.20% respectively. Direct AST performed on pediatric blood culture bottles using BD Phoenix M50 can quickly provide accurate susceptibility information to guide antimicrobial therapy in patients with BSI.

The Relationship Between Mortality and Leuko-Glycemic Index in Coronary Care Unit Patients (MORCOR-TURK LGI)

Introduction&amp;Objective: Identifying high-risk patients with a poor prognosis in coronary care unit (CCU) patients can assist physicians in providing optimal care and implementing preventive strategies. Leuko-glycaemic index (LGI), synthesized by multiplying the blood glucose level by the leukocyte count, has gained popularity in risk stratification of myocardial infarction patients. In this context, this study was carried out to investigate the relationship between LGI assessed at admission and in-hospital mortality in CCU patients. Methods: This is a multi-center, cross-sectional and observational study. (MORCOR-TURK LGI: Mortality Predictors in Coronary Care in Turkey, ClinicalTrials.gov number NCT05296694). The population of this study consisted of 2917 consecutive patients admitted to the CCU. Blood samples were collected into serum separator tubes in the immediate admission to the CCU. LGI was calculated by multiplying both values and dividing them by a thousand. LGI units were expressed in mg/dl. mm³. The sample was divided into two groups based on the LGI cut-off value of 1.23. Logistic regression analysis was used to find the significant predictors of mortality. Receiver operating characteristics (ROC) curve was to find out the cut-off value of LGI. A p value less than 0.05 was considered to be statistically significant in all analyses. Results: Univariable logistic regression analysis revealed that age, heart failure (HF), LGI, coronary artery disease, hypertension, diabetes mellitus and atrial fibrillation are clinically and statistically significant predictors. Further analysis of these variables using the multivariable logistic regression analysis indicated that age (Odds Ratio [OR]: 1.040, 95% confidence interval [CI]: 1.017-1.063; p=0.001), HF (OR: 2.426, 95% CI: 1.419-4.149; p:0.001) and LGI (OR: 1.349, 95% CI: 1.176-1.549; p3.72 predicted in-CCU mortality with 95.56% sensitivity and 49.19% specificity ([AUC]: 0.659 [95% CI: 0.641–0.676, p

Comparison of the Direct Identification and Short-Term Incubation Methods for Positive Blood Cultures via MALDI-TOF Mass Spectrometry.

Timely pathogen identification in bloodstream infections is crucial for patient care. A comparison is made between positive blood culture (BC) pellets from serum separator tubes using a direct identification (DI) method and colonies on agar plates from a short-term incubation (STI) method with a matrix-assisted laser desorption/ionization Biotyper for the evaluation of 354 monomicrobial BCs. Both the DI and STI methods exhibited similar identification rates for different types of bacteria, except for Gram-positive and anaerobic bacteria. The DI method's results aligned closely with the STI method's results for Enterobacterales, glucose-non-fermenting Gram-negative bacilli (GNB), and carbapenem-resistant Enterobacterales. The DI method exhibited high concordance with the conventional method for GNB identification, achieving 88.2 and 87.5% accuracy at the genus and species levels, respectively. Compared with the STI method, the DI method showed a less successful performance for Gram-positive bacterial identification (50.5 vs. 71.3%; p < 0.01). The DI method was useful for anaerobic bacterial identification of slow-growing microorganisms without any need for colony growth, unlike in the STI method (46.7 vs. 13.3%; p = 0.04). However, both methods could not identify yeast in positive BCs. Overall, the DI method provided reliable results for GNB identification, offering many advantages over the STI method by significantly reducing the turnaround time and enabling quicker pathogen identification in positive BCs.

Sample suitability and stability in different blood collection tubes for methanol, ethanol, isopropanol, acetone and glycols analysis

AimTo systematically examine the sample suitability and stability for volatile alcohols (methanol, ethanol and isopropanol with its metabolite acetone) and glycols (ethylene/propylene glycols, EG/PG) in different collection tubes.MethodTwo pools of...

Self-Sampling by Adolescents at Home: Assessment of the Feasibility to Successfully Collect Blood Microsamples by Inexperienced Individuals.

Blood microsampling has increasingly attracted interest in the past decades as a more patient-centric sampling approach, offering the possibility to collect a minimal volume of blood following a finger or arm prick at home. In addition to conventional dried blood spots (DBS), many different devices allowing self-sampling of blood have become available. Obviously, the success of home-sampling can only be assured when (inexperienced) users collect samples of good quality. Therefore, the feasibility of six different microsampling devices to collect capillary blood by inexperienced adolescents at home was evaluated. Participants (n = 95) were randomly assigned to collect blood (dried or liquid) at different time points using four of six different self-sampling devices (i.e., DBS, Mitra volumetric absorptive microsampling (VAMS), Capitainer B, Tasso M20, Minicollect tube and Tasso+ serum separator tube (SST)). The quality of the samples was visually inspected and analytically determined. Moreover, the participants' satisfaction was assessed via questionnaires. Although a majority succeeded based on the visual inspection, the success rate differed largely between the different devices. In general, the lowest success rate was obtained for the Minicollect tubes, although there is an opportunity and need for improvement for the other self-sampling devices as well. Hence, this also emphasizes the importance to assess the quality of samples collected by the target population prior to study initiation. In addition, visual classification by a trained individual was confirmed based on assessment of the analytical variability between replicates. Finally, self-sampling at home was overall (very) positively received by the participants.

The Impact of Reporting the Same-Day Identification and Antibiotic Susceptibility Test Results on the Treatment of Bloodstream Infections.

The rise of antibiotic-resistant organisms necessitates the implementation of rapid identification (ID) and antibiotic susceptibility testing (AST) methods for patient management. We aimed to analyze how rapid ID and AST reporting influenced clinicians' treatment decisions. Bacteria were identified directly from positive blood cultures (BC) using serum separator tubes and MALDI-TOF MS. EUCAST rapid antibiotic susceptibility testing (RAST) method was performed for AST. The impact of rapid ID and AST reports on clinician treatment decisions was evaluated through clinical documentation. The appropriateness of antimicrobial therapy and interventions was assessed according to institutional antimicrobial prescribing guidelines, AST results, and clinical data. A total of 128 BC bottles from 86 patients underwent processing. The rapid ID method was successful in 105 (82.1%) bottles obtained from 76 patients. The rapid ID results were reviewed by the Infectious Diseases Team on the same day for 55 (72.4%) of the 76 patients. Following the evaluation, new treatments or interventions were recommended for 28 (36.8%) patients. RAST results were available for 24 patients. The susceptibility profile of seven patients was assessed by the Infectious Diseases Team on the same day. Antimicrobial treatment was escalated in four cases, and de-escalation was made in two based on RAST results. If all rapid results had been assessed, adjustments could have been made for eight (10.5%) and eleven (14.5%) more patients, according to ID and RAST results, respectively. Implementation of rapid ID and AST may contribute to patient management. Although rapid reporting was made, some results were not evaluated by the clinician on the same day, indicating that communication between the clinician and the laboratory needs to be strengthened.

End-user verification results of two serum separator tubes for clinical chemistry analytes according to CLSI GP34-A and CLSI GP41-A6

Tube manufacturers use different composition of gels and blood clot activator formulations in serum tube production. Our aim was to investigate the within-tube (repeatability) and between-tube variation, concordance between comparison results of BD and VacuSEL tubes. Blood samples were collected from control subjects (n = 20) and patients (n = 30) in accordance with the CLSI GP41-A6 and CLSI GP34-A guidelines. Twenty-three clinical chemistry parameters were analysed via Roche Cobas C702 Chemistry Analyzer on T0 (0 hour) and T24 (24 hour). Mean differences % were compared with Wilcoxon matched pair test. Clinical significance was evaluated based on desirable bias according to total allowable error (TEa). VacuSEL tubes demonstrated acceptable performance for the results of 20 parameters with regards to desirable bias % limits. Lactate dehydrogenase (LD) [mean difference % (%95 confidence intervals (CI) values of BD and VacuSEL tubes at T0 [6.41% (4.80–8.01%)]; sodium (Na) and total protein (TP) at T24 [-0.27% (-0.46 to −0.07%) and −1.39% (-1.87 to −0.91), respectively] were over the desirable bias limits (LD: 4.3%, Na: 0.23% and TP: 1.36%, respectively) but not exceeding total biological variation CV % [Na: 0.5 (0.0–1.0) % and TP: 2.6 (2.3–2.7) %). %95 confidence intervals (CI) of T0 LD values overlap with within-subject biological variation % (CI) limits (LD: 5.2 (4.9–5.4) %). The differences between two tubes were not medically significant and necessarily conclusive. VacuSEL serum tubes presented comparable performance with BD serum tubes.

Emerging need of molecular profiling in hepatobiliary cancer

: Gallbladder cancer is a rare malignancy but represents almost 50% of all biliary tract cancer. Biliary cancers are highly fatal malignancies with a 5-year survival rate of approximately 20%. The prognosis of gallbladder cancer is poor due to the aggressive tumor biology, late presentation, complicated anatomic position, and advanced stage at diagnosis. Locally advanced and metastatic disease treatment is with palliative chemotherapy. Alarming sign of gallbladder cancer is overall decreased in incidence in older patients but increased in the younger population. So many mutations have been reported for the gall bladder cancer till date. : A prospective observational study was conducted over a period of 1 year at Asian Institute of Medical Sciences Faridabad which includes hepatobiliary carcinoma patients who are at stage III and stage IV of cancer. After getting the consent formalin fixed paraffin embedded biopsy samples, and 5 ml serum sample was collected in serum separator tube (SST). A whole genome sequencing was performed using Illumina HiSEQ, Illumina (NGS) technology, allows for high-throughput sequencing of DNA and RNA. Illumina's NGS is based on "sequencing by Synthesis" to detect the mutations.: Most common mutation found was in the P53 gene. TP53 (p.Arg175His), TP53 (p.Arg306Ter), TP53 (p.Cys238Tyr), TP53 (p.Leu43Ter), TP53 (p.Glu339Ter), TP53 (p.Pro190Leu). Mutations in the TP53 gene are a common feature of carcinoma of the gallbladder, and are associated with a more aggressive tumor phenotype, resistance to chemotherapy, and poorer overall survival.

Five days serum glucose stability at room-temperature in centrifuged fast-clotting serum tubes and the comparability with glucose in heparin-plasma and plasma containing citrate-stabilizer

Glucose measurement plays a central role in the diagnosis of gestational diabetes mellitus (GDM). Because of earlier reports of overestimation of glucose in the widely used tubes containing granulated glycolysis inhibitor, the study assessed the performance of fast-clotting serum tubes as an alternative sample for the measurement of glucose. Glucose concentration in fast-clotting serum was compared to lithium-heparin plasma placed in an ice-water slurry after sample collection and glucose stability at room-temperature was studied. Blood samples from 30 volunteers were drawn in four different types of tubes (serum separator tubes, fast-clotting serum tubes, lithium-heparin tubes and sodium fluoride, EDTA and a citrate buffer (NaF–EDTA–citrate) tubes, all from Greiner Bio-One). Lithium-heparin tubes were placed in an ice-water slurry until centrifugation in accordance with international recommendations and centrifuged within 10 min. After centrifugation, glucose was measured in all tubes (timepoint T 0) and after 24, 48, 72, 96 and 120 h of storage at 20–22 °C. NaF–EDTA–citrate plasma showed significant overestimation of glucose concentration by 4.7% compared to lithium-heparin plasma; fast-clotting serum showed glucose concentrations clinically equivalent to lithium-heparin plasma. In fast-clotting serum tubes, mean bias between glucose concentration after 24, 48, 72, 96 and 120 h and T 0 was less than 2.4%. All individual differences compared to T 0 were less than 6.5%. The results fulfill the acceptance criteria for sample stability based on biological variation. Fast-clotting serum tubes can be an alternative for the measurement of glucose in diagnosis and management of GDM and diabetes mellitus, especially when prolonged transportation is necessary.

Whole Blood Cardiac Troponin "Triaging" to Improve Early Detection of Myocardial Injury at a Pediatric Hospital.

The importance of offering on-site cardiac troponin (cTn) testing at pediatric hospitals may be underappreciated. We developed a rapid rule-in process for myocardial injury at a pediatric hospital experiencing delays in off-site high-sensitivity cardiac troponin T (hs-cTnT) testing. Collect-to-verify turnaround times (TATs) for off-site testing were reviewed. Pre-analytic changes to improve TATs were devised, implemented and evaluated, after which a new analyzer was selected and evaluated for on-site cTn testing. Performance of the new analyzer's assay was compared to the off-site hs-cTnT assay, and post go-live TATs for on-site testing were assessed. Median collect-to-verify TAT for short turnaround-time (STAT) priority off-site plasma hs-cTnT testing was 104 min, with 35% of orders having a TAT >120 min. Eliminating serum separator tubes and requiring a separate plasma separator tube did not significantly reduce TATs. A QuidelOrtho Triage® MeterPro whole blood cardiac troponin I (cTnI) assay was implemented to "triage" time-critical and STAT priority specimens collected for off-site hs-cTnT testing. Elevated cTnI (≥0.02 µg/L) had a sensitivity of 91% for clear elevations in hs-cTnT (≥53 ng/L) but a 0% sensitivity for modest elevations (5 to 13 ng/L, 14 to 52 ng/L). An interpretive comment was auto-appended to cTnI results indicating that clinicians should wait for the hs-cTnT result if cTnI was normal. Median collect-to-verify TAT for on-site cTnI testing was <50% the TAT for off-site hs-cTnT testing. On-site point-of-care whole blood cTn testing can rapidly confirm significant or late-presenting myocardial injury. Combined with simultaneous off-site high-sensitivity cardiac troponin (hs-cTn) testing, this workflow is a viable interim solution for pediatric hospitals without on-site hs-cTn testing.

P565 Capillary blood based self-collection device for infliximab therapeutic drug monitoring

Abstract Background Patient friendly new devices may improve therapeutic drug monitoring (TDM) in inflammatory bowel disease (IBD). Our objective was to evaluate an infliximab (IFX) precision guided dosing tool from capillary blood collected using an upper-arm self-collection device amongst IBD patients and compare its performances to gold standard venous blood collected using venipuncture. Methods Patients under maintenance therapy with IFX were recruited from a single IBD center and enrolled in the BEST (Bayesian Estimates Sustaining Trough) study. BEST is a prospective real world evidence clinical utility study to establish the value of an IFX Bayesian forecasting tool in the optimal management of IBD. Blood specimens were collected prospectively, at midcycle or at trough. Capillary blood specimens were collected using an upper arm self-collection device (Tasso Inc., Seattle, WA) connected to a serum separator tube. Paired venous blood was collected using venipuncture. IFX concentrations and antibodies to IFX (ATI) status were measured using drug tolerant homogenous mobility shift assay, albumin levels were measured using nephelometry. IFX Clearance was calculated using nonlinear mixed effect model with Bayesian priors. Statistical analysis consisted of Deming regression. Results Overall, 27 patients (n=9 with UC, mean age=37 years, weight=79 Kg) were enrolled. Median IFX dosing was 10 mg/Kg, and median dosing frequency every 6 weeks. A total of n=58 serum pairs isolated from capillary and venous blood were tested. PK profiles measured from venous blood were optimal during maintenance with no immune response detected (ATI &amp;lt; 3.1 U/mL) and most patients had IFX concentration at trough above 5 µg/mL (98%, median 15.4 µg/mL IQR: 10.9-28.0 µg/mL) and low Clearance (&amp;lt; 0.294 L/day: 89%; median 0.200 L/day IQR: 0.170-0.248 L/day). IFX concentrations and Clearance levels were comparable between venous and capillary blood with high correlation (R2= 0.975; slope =0.928; and R2=0.978; slope =1.035, respectively) (figure). In capillary blood specimens collected at midcycle (median 14 days before the next infusion, n=12 pairs), median IFX concentration was 38.3 µg/mL (IQR: 31.2-64.7 µg/mL) and declined to 21.9 µg/mL (IQR: 14.1-44.5 µg/mL) at trough. There was a high correlation between forecasted and measured trough concentrations in capillary blood (R2=0.925; Deming’s slope=1.093). Similar results were observed with venous blood pairs (R2=0.983; Deming’s slope=1.024). Conclusion IFX therapeutic drug monitoring using patient self-collection of capillary blood performed as well as standard venous blood measurement. This collection method may facilitate the implementation of point of care TDM in clinical practice.

Comparative and stability study of glucose concentrations measured in both sodium fluoride and serum separator tubes

IntroductionSodium fluoride/potassium oxalate (NaF/KOx) tubes has been regarded as the gold-standard tubes for glucose analysis. Even though their ineffectiveness in immediately inhibiting glycolysis has been reported in several studies especially in the first 1–4h, they are still used in our clinical biochemistry laboratory for glucose measurement. However, in its absence, only serum separator tubes are employed for glucose measurement. We aim to determine whether serum separator tubes (SSTs) can replace NaF/KOx tubes for laboratory measurement of blood glucose and to assess the stability of glucose concentrations for 3 days period. Methods and findingsNaF/KOx tube type was the reference method while SSTs type was the candidate method for glucose measurement. A total of 50 paired samples collected separately in NaF/KOx tubes and SSTs from healthy adult participants in the Gambia Adults Reference Intervals Study (GARIS) project were used as the project sample size. Following blood collection and separation, the glucose concentration was measured within 2 h, and at 24h, 42h and 72h time-points. Our data analysis showed no significant difference in the mean glucose concentrations between the reference tube and candidate tube types (Mean difference = 0.06 mmol/L; P = 0.38) recorded in the different timepoints. Using growth trajectory and mixed effects model, the study data further showed no significant change in the glucose concentrations (p = 0.25) for three days period. ConclusionsThe study confirms that SSTs can produce similar glucose results when employed in the absence of NaF/KOx tubes. Besides, the glucose concentrations were stable in both tubes for three days when the samples were separated within 2 h and refrigerated in 2–8°C.

Comparison of Various Blood Vacuum Tubes Brand for Clinical Chemistry, Hormonal, Hematology, and Hemostasis Parameters

Link of Video Abstract: https://youtu.be/O7-fCHyvGo4 Background: Laboratory tests are integral to the medical decision-making process. Blood vacuum tubes need to be validated to increase the analytics quality. This study aimed to compare various vacuum tube brands that use serum clot activator and serum separator tube (SST) for clinical chemistry and hormonal parameters, K3EDTA anticoagulant for hematological parameters, and citrate for hemostasis parameters. Methods: The comparative study was carried out cross-sectional in the Department of Clinical Pathology, Dr. Cipto Mangunkusumo National Public Hospital, on June 20, 2020. There were 20 subjects for each type of tube. The control tube was BD Vacutainer®, and the test tubes were Samplix®, VACUETTE®, Vacusera®, and YCHD. Results: Underfilling was the most common problem in preanalytical observation. This problem for citrate tubes was most frequently found in Vacusera® tubes. The possible cause of underfilling is the expiration date, which was close to the date of the experiment. There was a significant difference in PT results (p &lt;0.0001) in the Vacusera® citrate tube. All tube brands had no significant differences in all clinical chemistry, hormonal, and hematological parameters (P&gt;0.05). After 24 hours at room temperature, the results of hematological parameters showed statistically significant differences in MCV and hematocrit parameters, while other parameters were still within the CV limit of the instrument. Conclusion: There were no significant differences in the results of clinical chemistry, hormonal, and hematological parameters for all tube brands. A hematology parameters test should be done immediately as the delay may affect the MCV and hematocrit parameters.