Integrity of serum samples is changed by modified centrifugation conditions

2019 ◽  
Vol 57 (12) ◽  
pp. 1882-1887
Author(s):  
Marijana Miler ◽  
Nora Nikolac Gabaj ◽  
Jelena Culej ◽  
Adriana Unic ◽  
Alen Vrtaric ◽  
...  
Keyword(s):  
Turnaround Time ◽  
Serum Samples ◽  
P Values ◽  
H Index ◽  
Sample Quality

Abstract Background Serum samples should be centrifuged for at least 10 min at 1300–2500 × g. Changed centrifugation conditions could compromise sample quality. The objective of this study was to compare the serum quality and turnaround time (TAT) using different centrifugation conditions. Methods The study was done in four different periods (A, B, C and D) at different conditions: for 10, 5 and 7 (A, B and C, respectively) at 2876 × g, and 7 (D) min at 4141 × g. Sample quality was assessed as the proportion of samples with: (a) aspiration errors, (b) H index >0.5 g/L and (c) suppressed reports of potassium (K) due to hemolysis. TAT was calculated for emergency samples. The proportions of samples (a), (b) and (c) were compared according to period A. Results The number of aspiration errors was significantly higher in samples centrifuged at 2876 × g for 5 min (p = 0.021) and remained unchanged when centrifuged for 7 min (p = 0.066 and 0.177, for periods C and D, respectively). In periods B, C and D, the proportion of samples with hemolysis was higher than that in period A (p-values 0.039, 0.009 and 0.042, respectively). TAT differed between all periods (p < 0.001), with the lowest TAT observed for B and D. The lowest number of samples exceeding 60-min TAT was observed in period D (p = 0.011). Conclusions The integrity of serum samples is changed with different centrifugation conditions than those recommended. Our study showed that shorter centrifugation at higher force (7 min at 4141 × g) significantly decreases TAT, with unchanged proportion of samples with aspiration errors.

Automation and validation of a MALDI-TOF MS (Mass-Fix) replacement of immunofixation electrophoresis in the clinical lab

2021 ◽  
Vol 59 (1) ◽  
pp. 155-163
Author(s):  
Mindy Kohlhagen ◽  
Surendra Dasari ◽  
Maria Willrich ◽  
MeLea Hetrick ◽  
Brian Netzel ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Turnaround Time ◽  
Automated System ◽  
Serum Samples ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Automated Method ◽  
Positive Specimen ◽  
Specimen Identification ◽  
Tof Ms

AbstractObjectivesA matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) method (Mass-Fix) as a replacement for gel-based immunofixation (IFE) has been recently described. To utilize Mass-Fix clinically, a validated automated method was required. Our aim was to automate the pre-analytical processing, improve positive specimen identification and ergonomics, reduce paper data storage and increase resource utilization without increasing turnaround time.MethodsSerum samples were batched and loaded onto a liquid handler along with reagents and a barcoded sample plate. The pre-analytical steps included: (1) Plating immunopurification beads. (2) Adding 10 μl of serum. (3) Bead washing. (4) Eluting the immunoglobulins (Igs), and reducing to separate the heavy and light Ig chains. The resulting plate was transferred to a second low-volume liquid handler for MALDI plate spotting. MALDI-TOF mass spectra were collected. Integrated in-house developed software was utilized for sample tracking, driving data acquisition, data analysis, history tracking, and result reporting. A total of 1,029 residual serum samples were run using the automated system and results were compared to prior electrophoretic results.ResultsThe automated Mass-Fix method was capable of meeting the validation requirements of concordance with IFE, limit of detection (LOD), sample stability and reproducibility with a low repeat rate. Automation and integrated software allowed a single user to process 320 samples in an 8 h shift. Software display facilitated identification of monoclonal proteins. Additionally, the process maintains positive specimen identification, reduces manual pipetting, allows for paper free tracking, and does not significantly impact turnaround time (TAT).ConclusionsMass-Fix is ready for implementation in a high-throughput clinical laboratory.

scholarly journals Validation of the Procalcitonin Assay on the Abbott Architect i1000

2019 ◽  
Vol 3 (6) ◽  
pp. 936-942 ◽  
Author(s):  
Jayson V Pagaduan ◽  
Estella Tam ◽  
Sridevi Devaraj
Keyword(s):  
Statistical Analysis ◽  
Method Validation ◽  
Turnaround Time ◽  
Good Precision ◽  
Serum Samples ◽  
Diagnostic Use ◽  
Clinical Laboratories ◽  
The Us ◽  
The Impact ◽  
Abbott Architect

Abstract Background Procalcitonin (PCT) is an emerging biomarker for detecting sepsis. Recently, the US Food and Drug Administration cleared the expanded use of this biomarker for guiding clinicians regarding antibiotic treatment. To our knowledge, there are no published method validations for the Abbott Architect PCT assay. This article will discuss the process of method validation of the B·R·A·H·M·S PCT assay on the Abbott Architect platform. Methods We studied the precision, accuracy, and linearity of the Architect method following the guidance of the Clinical and Laboratory Standards Institute EP5-A2 document. Furthermore, we also tested the impact of major sources of interference from hemolysate, lipoproteins, and bilirubin. To validate the Architect method, we compared patients' serum PCT measurements with our previously established Mini VIDAS (bioMerieux) PCT assay. Results Statistical analysis showed that the 2 assays have good correlation (r &gt; 0.99), slope of 1.023, and intercept of −0.760. The calculated bias is −7.435%. The Architect method showed good precision with %CV &lt; 3.5% for both interassay and intraassay compared with %CV &lt; 6.5% for Mini VIDAS, which was previously determined at our institution. No bias &gt;10% was observed with the Architect method when pooled serum samples were spiked with interferants. The turnaround time for both platforms was the same (20 min); however, in contrast with Mini VIDAS, the Architect system has automated pipetting of samples and can perform multiple assays simultaneously. Conclusion These results showed that the Architect B·R·A·H·M·S PCT assay has analytical characteristics conducive for diagnostic use in clinical laboratories. Our method validation report will be beneficial for other institutions to adapt this assay on existing Abbott Architect i1000 immunoassay analyzers.

scholarly journals Point-of-care high-sensitivity assay on PATHFAST as the backup in the emergency room

2021 ◽  
Vol 5 ◽  
pp. 239920262110550
Author(s):  
Joško Osredkar ◽  
Katja Krivic ◽  
Teja Fabjan ◽  
Kristina Kumer ◽  
Jure Tršan ◽  
...  
Keyword(s):  
Blood Serum ◽  
Troponin I ◽  
Point Of Care ◽  
High Sensitivity ◽  
Turnaround Time ◽  
Serum Samples ◽  
Linear Relationships ◽  
Accuracy And Precision ◽  
Laboratory Results ◽  
Key Indicators

Aim: Although the levels of cardiac troponin I (cTnI) have proved to be a useful diagnostic biomarker of acute myocardial infarction, there are a wide variety of point-of-care (POC) analysers, which provide measurements of cTnI. The aim of this study was to compare the results obtained by the ADVIA Centaur ultra-assay cTnI assay (us-cTnI), ADVIA Centaur high-sensitive cTnI assay (hs-cTnI) and a POC high-sensitivity assay using PATHFAST. We also aimed to explore total turnaround time (TAT) for laboratory results using the POC PATHFAST analyser. Methods: Samples from 161 patients were taken. Of these samples, 129 were tested with all three assays (us-cTnI, hs-cTnI and PATHFAST), and 32 samples were tested on PATHFAST for the comparison of whole blood, serum and plasma. Results: Comparison of the POC testing methods in this study demonstrated that there are strong linear relationships between all three cTnI assays (us-cTnI, hs-cTnI and POC on PATHFAST). Furthermore, we also show there are strong linear relationships between the two high-sensitive cTnI assays (hs-cTnI and PATHFAST) for blood serum samples, as determined by Passing–Bablok regression analyses. In our comparison of our new data with our older study, the TAT went down. Conclusion: The timeliness of laboratory results is, in addition to accuracy and precision, one of the key indicators of laboratory performance, and at the same time has a significant impact on the course of the patient’s condition. It is therefore important that the laboratory strives to meet the expectations of clinicians regarding the time from the order to the result of the analysis.

Evaluation of two point of care technologies for measuring monoclonal antibody therapeutic-A concentrations in blood

Bioanalysis ◽  
2020 ◽  
Vol 12 (20) ◽  
pp. 1449-1458
Author(s):  
Saloumeh K Fischer ◽  
Kathi Williams ◽  
Ian Harmon ◽  
Bryan Bothwell ◽  
Hua Xu ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Real Time ◽  
Point Of Care ◽  
Turnaround Time ◽  
Monitoring Methods ◽  
Sample Collection ◽  
Time Data ◽  
Serum Samples ◽  
Quick Turnaround Time

Aim: Current blood monitoring methods require sample collection and testing at a central lab, which can take days. Point of care (POC) devices with quick turnaround time can provide an alternative with faster results, allowing for real-time data leading to better treatment decisions for patients. Results/Methodology: An assay to measure monoclonal antibody therapeutic-A was developed on two POC devices. Data generated using 75 serum samples (65 clinical & ten spiked samples) show correlative results to the data generated using Gyrolab technology. Conclusion: This case study uses a monoclonal antibody therapeutic-A concentration assay as an example to demonstrate the potential of POC technologies as a viable alternative to central lab testing with quick results allowing for real-time decision-making.

scholarly journals IL8 and IL16 levels indicate serum and plasma quality

2018 ◽  
Vol 56 (7) ◽  
pp. 1054-1062 ◽  
Author(s):  
Olga Kofanova ◽  
Estelle Henry ◽  
Rocio Aguilar Quesada ◽  
Alexandre Bulla ◽  
Hector Navarro Linares ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Room Temperature ◽  
Plasma Samples ◽  
Sample Processing ◽  
Serum Samples ◽  
Sample Quality ◽  
Edta Plasma ◽  
Citrate Plasma

Abstract Background: Longer pre-centrifugation times alter the quality of serum and plasma samples. Markers for such delays in sample processing and hence for the sample quality, have been identified. Methods: Twenty cytokines in serum, EDTA plasma and citrate plasma samples were screened for changes in concentration induced by extended blood pre-centrifugation delays at room temperature. The two cytokines that showed the largest changes were further validated for their “diagnostic performance” in identifying serum or plasma samples with extended pre-centrifugation times. Results: In this study, using R&D Systems ELISA kits, EDTA plasma samples and serum samples with a pre-centrifugation delay longer than 24 h had an IL16 concentration higher than 313 pg/mL, and an IL8 concentration higher than 125 pg/mL, respectively. EDTA plasma samples with a pre-centrifugation delay longer than 48 h had an IL16 concentration higher than 897 pg/mL, citrate plasma samples had an IL8 concentration higher than 21.5 pg/mL and serum samples had an IL8 concentration higher than 528 pg/mL. Conclusions: These robust and accurate tools, based on simple and commercially available ELISA assays can greatly facilitate qualification of serum and plasma legacy collections with undocumented pre-analytics.

A rapid, sensitive enzyme-linked immunoassay for human thyrotropin.

1985 ◽  
Vol 31 (7) ◽  
pp. 1131-1134 ◽  
Author(s):  
Y C Tseng ◽  
K D Burman ◽  
J R Baker ◽  
L Wartofsky
Keyword(s):  
Color Change ◽  
Clinical Laboratory ◽  
High Sensitivity ◽  
Normal Subjects ◽  
Turnaround Time ◽  
Cross Reactivity ◽  
Serum Samples ◽  
Assay Sensitivity ◽  
Nitrophenyl Phosphate ◽  
Set Up

Abstract In this enzyme-linked immunoassay for human thyrotropin (TSH) in unextracted serum we use 96-well immunoenzymometric assay plates, first coated with polyclonal antibody to TSH, then incubated with the serum samples and reacted with mouse monoclonal antibody to human TSH. After incubation with alkaline phosphatase-labeled antibody against mouse IgG, disodium p-nitrophenyl phosphate is added and the color change is measured spectrophotometrically. Assay sensitivity is 0.1 milli-int. unit/L. Cross reactivity with lutropin, follitropin, or choriogonadotropin was negligible. TSH concentrations ranged from 0.4 to 4.1 milli-int. units/L in 43 normal subjects (mean 2.0, SD 1.0), and were uniformly less than 0.3 milli-int. unit/L in 23 patients with hyperthyroidism. Features which make this assay advantageous to the clinical laboratory include ease of set-up, ability to assay many samples at a time, high sensitivity, rapid turnaround time (8 h), and absence of requirements for radioactive materials.

scholarly journals High-Quality, Cost-Effective Strategy for Detection of Autoantibodies to Extractable Nuclear Antigens

2001 ◽  
Vol 8 (3) ◽  
pp. 471-474 ◽  
Author(s):  
Tri G. Phan ◽  
Watson W. S. Ng ◽  
Dana Bird ◽  
Kara Smithers ◽  
Vicky Wong ◽  
...  
Keyword(s):  
Cost Effective ◽  
Turnaround Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Quality Cost ◽  
Nuclear Antigens ◽  
Cost Effective Method ◽  
Meaningful Result ◽  
Clinical Diagnoses

ABSTRACT We evaluated methods for the detection of autoantibodies to extractable nuclear antigens (ENAs) to determine the strategy that yielded the most cost effective and clinically meaningful result. We prospectively compared counterimmunoelectrophoresis (CIEP) with and without serum prediffusion (SPD) and found that SPD significantly improved the quality of precipitation lines. This resulted in a decreased requirement for repeat testing and, consequently, was associated with a significant decrease in reagent costs and specimen turnaround time. We also retrospectively compared reactivity by CIEP, CIEP plus SPD, enzyme-linked immunosorbent assay (ELISA), and line immunoassay (LIA) of 52 serum samples that were previously determined to be positive for ENAs, and we correlated the results with clinical diagnoses. There was significant agreement among CIEP, CIEP plus SPD, ELISA, and LIA for the detection of anti-SS-A, anti-SS-B and anti-RNP. In general, CIEP, CIEP plus SPD, and LIA correlated better with the clinical diagnoses than ELISA, even though ELISA detected anti-ENAs more often than the other methods. CIEP plus SPD is therefore the most cost effective method for the identification of clinically meaningful ENAs. Based on our experience, we now screen for ENAs by CIEP, and positive samples are then typed by CIEP plus SPD. Samples that are difficult to interpret are then further assessed by an alternative method.

scholarly journals Pre-Analytical Sample Quality: Metabolite Ratios as an Intrinsic Marker for Prolonged Room Temperature Exposure of Serum Samples

PLoS ONE ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. e0121495 ◽  
Author(s):  
Gabriele Anton ◽  
Rory Wilson ◽  
Zhong-hao Yu ◽  
Cornelia Prehn ◽  
Sven Zukunft ◽  
...  
Keyword(s):  
Room Temperature ◽  
Serum Samples ◽  
Sample Quality ◽  
Intrinsic Marker ◽  
Temperature Exposure

scholarly journals Improved Sample Quality and Decreased Turnaround Time When Using Plasma Blood Collection Tubes with a Mechanical Separator in a Large University Hospital

2020 ◽  
Author(s):  
Pierre-Olivier Hétu ◽  
Sacha Hobeila ◽  
François Larivière ◽  
Marie-Claire Bélanger
Keyword(s):  
Clinical Chemistry ◽  
Turnaround Time ◽  
Error Rates ◽  
University Hospital ◽  
Blood Collection ◽  
Sample Quality ◽  
Paired Samples ◽  
Deming Regression ◽  
Before And After ◽  
The Impact

Abstract Background Serum is commonly used for clinical chemistry testing but many conditions can affect the clotting process, leading to poor sample quality and impaired workflow. With serum gel tubes, we found a high proportion of sample probe aspiration errors on our Beckman AU5800 analyzers. We decided to implement the BD Barricor™ plasma tubes, and we validated an off-specification centrifugation scheme and verified that results obtained for 65 chemistry and immunochemistry tests were comparable to those obtained in serum gel tubes. Finally, we evaluated the impact of this new tube on sample error rate and laboratory turnaround time. Methods To validate centrifugation settings, 50 paired samples were collected in Barricor tubes and centrifuged at 1912 × g for 10 min or 5 min (off-specification). To compare serum gel tubes with Barricor plasma tubes, 119 paired samples were collected from volunteers and results were analyzed using weighed Deming regression. Finally, the proportion of aspiration errors and laboratory TAT for potassium were measured before and after implementing Barricor tubes. Results Barricor tubes showed clinically acceptable equivalence to serum gel tubes for the studied analytes, and the off-specification centrifugation scheme did not affect the results. Implementing Barricor tubes improved the laboratory workflow by decreasing the aspiration error rates (2.01% to 0.77%, P &lt; 0.001) and lowering hemolysis (P &lt; 0.001). The laboratory TAT for potassium were also significantly lowered (P &lt; 0.001). Conclusion Use of Barricor tubes instead of serum gel tubes leads to better sample quality, shorter more reproducible laboratory TAT, and decreases costs associated with error management.

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