Analytical performance of an automated volumetric flow cytometer for quantitation of T, B and natural killer lymphocytes
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Simon Degandt
Abstract
Background:
Quantitation of lymphocyte subsets (B cells, T cells, CD4 and CD8 T cells and NK cells) classically relies on quantitation of lymphocytes and immunophenotyping by flow cytometry. AQUIOS CL (Beckman Coulter) is a fully automated system that performs an onboard volumetric cell count, automatically processes the sample (staining, lysing and fixation) and analyzes the results. We compared AQUIOS CL to a dual-platform analysis and evaluated analytical performance.
Methods:
We evaluated precision, sample stability, inter-sample carryover, linearity and interpanel consistency. AQUIOS CL was compared to a dual-platform method (Sysmex XE-5000 and BD FACSCanto-II). A total of 113 patient samples were included: 45 from posttransplant patients, 44 from children and 24 random routine samples. The degree of automation was scored through the need of manual revisions triggered by AQUIOS CL run notifications and run flags.
Results:
Intrarun and interrun variability was <9.1% with dedicated control material and <32.1% with patient samples. Relative values of lymphocyte subsets could be determined up to 48 h after venipuncture when the sample was kept at room temperature. There was no carryover and good linearity. Interpanel consistency was 3.3% for relative values and 9.4% for absolute values. Method comparison showed good analytical correlation between AQUIOS CL and a dual-platform method. Thirty-five percent of the samples triggered a run notification. In 74% of these samples, the results could be accepted without intervention, so in 26% of all samples, an unnecessary notification was generated.
Conclusions:
AQUIOS CL allows for reliable fully automated immunophenotyping of lymphocyte subset quantitation. Gating algorithms could be further improved.
Acknowledgments
This study was supported by Beckman Coulter through donations of laboratory equipment and supplies. This private company had no role in study design and the collection of data. They did help in interpreting gating results with run notifications. Beckman Coulter had no role in the preparation of the manuscript or the decision to submit it for publication. We thank Professor David Barnett and Liam Whitby (UK National External Quality Assessment Scheme [UK NEQAS], Sheffield) for providing information and data regarding the UK NEQAS immune monitoring scheme.
Author contributions: BP, NB, XB: study design. SD: drafted the manuscript. BP, SJ: patient recruitment. SD, BP, SJ: data acquisition. SD, BP: statistical analysis. NB,XB checked and improved content and English grammar and style of the manuscript. NB, XB commented on and edited the manuscript, which was read and approved by all authors. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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Supplementary Material:
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2017-0638).
©2018 Walter de Gruyter GmbH, Berlin/Boston
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Artikel in diesem Heft
- Frontmatter
- Editorials
- Procalcitonin for diagnosing and monitoring bacterial infections: for or against?
- Is it time to abandon the Nobel Prize?
- In Memoriam
- Norbert Tietz, 13th November 1926–23rd May 2018
- Reviews
- Procalcitonin guidance in patients with lower respiratory tract infections: a systematic review and meta-analysis
- Telomere biology and age-related diseases
- Opinion Paper
- Procalcitonin-guided antibiotic therapy: an expert consensus
- Genetics and Molecular Diagnostics
- mRNA expression profile in peripheral blood mononuclear cells based on ADRB1 Ser49Gly and Arg389Gly polymorphisms in essential hypertension — a case-control pilot investigation in South Indian population
- General Clinical Chemistry and Laboratory Medicine
- External quality assessment schemes for glucose measurements in Germany: factors for successful participation, analytical performance and medical impact
- Long-term stability of glucose: glycolysis inhibitor vs. gel barrier tubes
- Observational studies on macroprolactin in a routine clinical laboratory
- Anti-streptavidin IgG antibody interference in anti-cyclic citrullinated peptide (CCP) IgG antibody assays is a rare but important cause of false-positive anti-CCP results
- Point-of-care creatinine testing for kidney function measurement prior to contrast-enhanced diagnostic imaging: evaluation of the performance of three systems for clinical utility
- Hematology and Coagulation
- Analytical performance of an automated volumetric flow cytometer for quantitation of T, B and natural killer lymphocytes
- Lupus anticoagulant testing using two parallel methods detects additional cases and predicts persistent positivity
- Reference Values and Biological Variations
- Within-subject biological variation of activated partial thromboplastin time, prothrombin time, fibrinogen, factor VIII and von Willebrand factor in pregnant women
- Within-subject and between-subject biological variation estimates of 21 hematological parameters in 30 healthy subjects
- Immunoglobulin G (IgG) and IgG subclass reference intervals in children, using Optilite® reagents
- Cancer Diagnostics
- Serum carbohydrate sulfotransferase 7 in lung cancer and non-malignant pulmonary inflammations
- Cardiovascular Diseases
- Clinical performance of a new point-of-care cardiac troponin I test
- Diabetes
- Testing for HbA1c, in addition to the oral glucose tolerance test, in screening for abnormal glucose regulation helps to reveal patients with early β-cell function impairment
- Effects of common hemoglobin variants on HbA1c measurements in China: results for α- and β-globin variants measured by six methods
- Infectious Diseases
- Serum procalcitonin concentration within 2 days postoperatively accurately predicts outcome after liver resection
- Predictive value of serum gelsolin and Gc globulin in sepsis – a pilot study
- Cerebrospinal fluid free light chains as diagnostic biomarker in neuroborreliosis
- Letter to the Editor
- Glucose and total protein: unacceptable interference on Jaffe creatinine assays in patients
- Interference of glucose and total protein on Jaffe-based creatinine methods: mind the covolume
- Interference of glucose and total protein on Jaffe based creatinine methods: mind the covolume – reply
- Measuring procalcitonin to overcome heterophilic-antibody-induced spurious hypercalcitoninemia
- Paraprotein interference in the diagnosis of hepatitis C infection
- Timeo apis mellifera and dona ferens: bee sting-induced Kounis syndrome
- Bacterial contamination of reusable venipuncture tourniquets in tertiary-care hospital
- Detection of EDTA-induced pseudo-leukopenia using automated hematology analyzer with VCS technology
- Early adjustment of antimicrobial therapy after PCR/electrospray ionization mass spectrometry-based pathogen detection in critically ill patients with suspected sepsis
- Thromboelastometry reveals similar hemostatic properties of purified fibrinogen and a mixture of purified cryoprecipitate protein components
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