K2-EDTA And K3-EDTA Greiner Tubes For HbA 1c Measurement

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Article Navigation Article Navigation Journal Article K2-EDTA and K3-EDTA Greiner Tubes for HbA1c Measurement Alen Vrtaric, MSc, Alen Vrtaric, MSc University Department of Chemistry, Medical School University Hospital Sestre Milosrdnice, Zagreb, Croatia *To whom correspondence should be addressed.alenvrtaric@gmail.com Search for other works by this author on: Oxford Academic PubMed Google Scholar Petra Filipi, MSc, Petra Filipi, MSc University Department of Chemistry, Medical School University Hospital Sestre Milosrdnice, Zagreb, Croatia Search for other works by this author on: Oxford Academic PubMed Google Scholar Marina Hemar, MSc, Marina Hemar, MSc University Department of Chemistry, Medical School University Hospital Sestre Milosrdnice, Zagreb, Croatia Search for other works by this author on: Oxford Academic PubMed Google Scholar Nora Nikolac, PhD, Nora Nikolac, PhD University Department of Chemistry, Medical School University Hospital Sestre Milosrdnice, Zagreb, Croatia Search for other works by this author on: Oxford Academic PubMed Google Scholar Ana-Maria Simundic, PhD Ana-Maria Simundic, PhD University Department of Chemistry, Medical School University Hospital Sestre Milosrdnice, Zagreb, Croatia Search for other works by this author on: Oxford Academic PubMed Google Scholar Laboratory Medicine, Volume 47, Issue 1, February 2016, Pages 39–42, https://doi.org/10.1093/labmed/lmv016 Published: 04 January 2016 Navbar Search Filter Laboratory MedicineThis issueASCP Journals PathologyBooksJournalsOxford Academic Mobile Enter search term Search Close Navbar Search Filter Laboratory MedicineThis issueASCP Journals PathologyBooksJournalsOxford Academic Enter search term Search Advanced Search Search Menu

ABSTRACT

Objective: To determine whether K2-ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Greiner tubes could be used interchangeably for glycosylated hemoglobin, type A1C (HbA1c) measurement via the Abbott Laboratories ARCHITECT chemiluminescent microparticle HbA1c assay on the ARCHITECT i2000SR immunoanalyzer at our university hospital.

Materials: We drew blood from a total of 45 outpatients into plastic Greiner Vacuette tubes, some of which were lined with K2-EDTA and others with K3-EDTA anticoagulant. Data are presented as median and interquartile range values. We used the Wilcoxon test and Passing-Bablok regression for tube comparison.

Results: For K2-EDTA tubes median HbA1c concentration was 54 mmol/mol (41 to 71 mmol/mol) and for K3-EDTA tubes 56 mmol/mol (43 to 69 mmol/mol). There was no statistically significant difference between K2-EDTA and K3-EDTA (bias= -1.29 mmol/mol; P = 0.24). Passing-Bablok regression showed that there is no constant and proportional error: y = ‐0.23 (95% CI[-3.52 to 0.69]) + 1.00( 95% CI[0.98 to 1.06]) x.

Conclusion: In this study, we provide evidence for the lack of any clinically and statistically significant bias between K2-EDTA and K3-EDTA HbA1c measurements. Thus, Greiner tubes lined with K2-EDTA and those lined with K3-EDTA can safely be used interchangeably to measure HbA1c via the Abbott Laboratories ARCHITECT assay.

HbA1c, preanalytical phase, method comparison, test tube comparison, K2-EDTA, K3-EDTA

Measurement of HbA1c is the criterion standard for therapy monitoring in patients with diabetes and a diagnostic test for diabetes mellitus, provided that stringent quality assurance is in place and its accurate measurement can be ensured.1,2 Current recommendation for glycosylated hemoglobin, type A1C (HbA1c) measurement imprecision is less than 3.0%, although even more stringent criteria have been proposed by some authors,3,4 namely, an analytical goal for imprecision of less than 2.0% and maximal allowable bias of ± 2.8%. To ensure such strict criteria, a laboratory must minimize all potential sources of preanalytical and analytical errors.

Blood tubes and blood tube components, such as anticoagulants, gels, and clot activators, have long been recognized as a significant source of variability, and laboratories are expected to provide data to prove that the tubes in use do not contribute to analytical interference.5,6 HbA1c measurement is usually performed in ethylenediaminetetraacetic acid (EDTA)–anticoagulated whole blood. EDTA comes in blood tubes as dipotassium, tripotassium, and sodium EDTA salt. It has been recognized7,8 that the choice of the type of EDTA salt may affect the accuracy of cell counting and sizing. Hence, the International Council for Standardization in Hematology (ICSH) recommends K2-EDTA as the anticoagulant of choice for hematology testing. Moreover, due to its ability to preserve cell morphology and integrity, EDTA has also been recommended as the anticoagulant of choice for other assays, such as cytokine, hormone and molecular biology testing, as well as for some clinical chemistry analytes.

In our laboratory, we use the ARCHITECT HbA1c Immunoassay (Abbott Laboratories). K2-EDTA, fluoride oxalate and sodium fluoride/potassium EDTA, as substances within whole-blood collection tubes, are declared by the manufacturer to be preferred anticoagulants for the HbA1c ARCHITECT Immunoassay. In our laboratory, we predominantly use K3-EDTA Greiner Vacuette EDTA tubes (Greiner Bio-One GmbH) coated with spray-dried K3-EDTA for other assays. However, K3-EDTA tubes were not declared by the manufacturer to be an acceptable choice of anticoagulant for the HbA1c immunoassay. Therefore, our aim was to assess whether K3-EDTA Greiner tubes introduce any significant bias in HbA1c measurement by the ARCHITECT HbA1c Immunoassay, compared with K2-EDTA tubes (the anticoagulant recommended by the manufacturer).

Materials and Methods

Subjects

Specimen collection was performed in the outpatient unit of the Clinical Institute of Chemistry, University Department of Chemistry at the University Hospital Center Sestre Milosrdnice, Zagreb, Croatia, between December 2014 and January 2015. Blood was drawn from outpatients (n = 45), who had been fasting for at least 8 hours, between 7:30 AM and 9:00 AM. For each patient, 2 plastic Vacuette tubes of 3 mL each were obtained: 1 lined with K2-EDTA and the other with K3-EDTA. This study was approved by the Hospital Ethics committee and was performed in accordance with the ethical principles of the Declaration of Helsinki. Laboratory staff asked all patients before blood was drown, only patients who gave oral consent participated in this study.

After specimen retrieval, we immediately mixed the tube contents 8 to 10 times, transported the tubes to the biochemistry laboratory, and remixed the tube contents another 8 to 10 times, according to manufacturer recommendation, immediately before analysis. As long as 1 hour passed from specimen retrieval to analysis. We measured HbA1c concentrations on the ARCHITECT i2000SR immunochemistry analyzer (Abbott Laboratories) using chemiluminescent microparticle immunoassay (CMIA) technology (HbA1c, ref. 4P72). Analytical imprecision was verified using Clinical and Laboratory Standards Institute (CLSI) EP5-A2 protocol.11 Because we aimed to detect possible differences between 2 specimens in cases in which measurements were performed consecutively, we have presented our data on repeatability: the coefficient of variation (CV) was 1.18% at 42 mmol/mol and 1.84% at 88 mmol/mol for HbA1c Lyphochek Diabetes controls (BioRad Laboratories, Inc.) Levels 1 and 2, respectively. The HbA1c results are reported in mmol/mol.

Statistical Analysis

We tested normality using the Kolmogorov-Smirnov test. Because distribution was not normal, we present our data as medians and interquartile ranges. Differences between K2-EDTA and K3-EDTA measurements were tested via the nonparametric Wilcoxon test for paired observations. Passing-Bablok regression analysis and Bland-Altman plotting were performed to assess the comparability of results obtained by 2 different additive types. We have selected ±2.8% as the maximal allowable bias between K2-EDTA and K3-EDTA tubes. P < 0.05 was considered statistically significant. MedCalc statistical software, version 12.7.2.0 (MedCalc Software, Ostend, Belgium) was used to analyze data.

Results

The HbA1c measurements in our study cover the entire measuring interval. The lowest measured concentration of HbA1c in our study was 21.3 mmol/mol, and the highest concentration was 117.5 mmol/mol. The observed mean difference between K2-EDTA and K3-EDTA was neither clinically nor statistically significant (P = 0.24). Passing-Bablok regression showed there is no constant and proportional bias between HbA1c measured in K2-EDTA and K3-EDTA tubes. Proportional bias between HbA1c measured in K2-EDTA and K3-EDTA tubes and the line of identity is superimposed on the regression line (Figure 1). Bland-Altman plotting also confirmed that the differences between 2 specimens were not statistically significant (Figure 2). Passing-Bablok regression analysis of the comparability of HbA1c concentrations measured in K2‐ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Vacuette Greiner (Greiner Bio-One GmbH) tubes. Solid line represents regression line; dashed lines represent confidence interval (CI) for the regression line; dotted line is the line of identity (x = y). y = ‐0.23 (95% CI[-3.52 - 0.69])+1.00 (95% CI[0.98 - 1.06]) x.Figure 1

Passing-Bablok regression analysis of the comparability of HbA1c concentrations measured in K2‐ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Vacuette Greiner (Greiner Bio-One GmbH) tubes. Solid line represents regression line; dashed lines represent confidence interval (CI) for the regression line; dotted line is the line of identity (x = y). y = ‐0.23 (95% CI[-3.52 - 0.69])+1.00 (95% CI[0.98 - 1.06]) x.

Open in new tabDownload slideBland-Altman plot of the comparability of HbA1c concentrations measured in K2-ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Vacuette Greiner tubes (Greiner Bio-One GmbH). The outer dotted lines represent limits of agreement (mean difference ±1.96 SD). Because confidence interval limits were at the opposite side of the zero (dotted lines), this difference is not statistically significant. The lines consisting of alternative dots and dashes represent where the bias value equals 0.Figure 2

Bland-Altman plot of the comparability of HbA1c concentrations measured in K2-ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Vacuette Greiner tubes (Greiner Bio-One GmbH). The outer dotted lines represent limits of agreement (mean difference ±1.96 SD). Because confidence interval limits were at the opposite side of the zero (dotted lines), this difference is not statistically significant. The lines consisting of alternative dots and dashes represent where the bias value equals 0.

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Discussion

Our results show that tubes coated with spray-dried K2-EDTA and K3-EDTA can be used interchangeably for HbA1c measurement, given the absence of statistical and clinical difference between K2 and K3 salts. To our knowledge, the comparability of K2-EDTA and K3-EDTA for HbA1c measurement has not yet been studied. Mailankot et al12 have studied the comparability of various anticoagulants (EDTA, sodium citrate, heparin, and fluoride) for HbA1c measurement. However, their study was performed with a sample of limited size (4 normal healthy men and 2 patients with diabetes). Moreover, the authors did not specify the type of potassium salt (K2-EDTA or K3-EDTA). Recently, Chakraborty and colleagues have published a study of the comparability of potassium EDTA and sodium fluoride/sodium EDTA Vacutainer tubes (Becton Dickinson and Company), and citrate-buffered fluoride sodium/EDTA tubes (Terumo Venosafe) for high-performance liquid chromatography (HPLC) measurement of HbA1c.13 Their results show that differences between tube types are lower than the acceptable clinically significant differences and, as such, are not likely to have any clinical implications. Unfortunately, the authors of this study have also failed to report the type of potassium salt used in their study. It is therefore not possible to compare their results with ours.

A potential limitation to our study might be that we did not confirm the presence of other hemoglobin variants that might influence the result of HbA1c measurement using the ARCHITECT CMIA method.14 Based on data from the manufacturer, interferences from hemoglobin HbC and HbS are less than 2.0% and 2.1%, respectively. The manufacturer lists possible interference from HbF; however, specimens where we expected elevated concentrations of HbF were excluded. Hemoglobins D and E are listed in the data from the manufacturer as presenting significant interference (bias values 28.8% and 21.1%, respectively).15 However, our study was designed to measure the difference in HbA1c concentration between 2 specimens from the same patient, divided into 2 different tubes. If any of these variants were present among our specimens, we believe that both tubes would be equally affected.

The use of K2-EDTA and K3-EDTA tubes in the same laboratory is not uncommon, especially nowadays, given laboratory consolidation and merger of laboratories from geographically wide areas. To minimize the risk of introducing bias due to interchangeable use of K2-EDTA and K3-EDTA tubes from different collection facilities, the central laboratory must ensure that potassium EDTA salts ensure comparable results. Our study provides evidence for the lack of any clinically and statistically significant bias between K2-EDTA and K3-EDTA HbA1c measurements. We hope that our results may serve other laboratories as they consider the optimal choice of tube and additive for HbA1c measurements, in cutting material costs and improving laboratory efficiency and patient safety.

We thank Greiner Bio-One GmbH, Kremsmünster, Austria, and Abbott Laboratories, Abbott Park, IL, for supporting this study by providing the EDTA tubes and HbA1c reagent free of charge.

References

1

World Health Organization. Use of Glycated Haemoglobin (HbA1c) in the Diagnosis of Diabetes Mellitus. WHO/NMH/CHP/CPM/11.1. Geneva, Switzerland: World Health Organization; 2011.

Google Scholar

Google Preview

OpenURL Placeholder Text

WorldCat

COPAC2

American Diabetes Association. http://care.diabetesjournals.org/content/38/Supplement_1. Accessed June 15, 2015. Diabetes care 2015;38(Suppl1.):S8-S16.

3

Braga F, Panteghini M. Standardization and analytical goals for glycated hemoglobin measurement. Clin Chem Lab Med. 2013;51:1719–1726.

Google Scholar

CrossrefSearch ADSPubMed

WorldCat

 4

Weycamp C. HbA1c: a review of analytical and clinical aspects. Ann Lab Med. 2013;33:393–400.

Google Scholar

CrossrefSearch ADSPubMed

WorldCat

 5

Bowen RAR, Remaley TA. Interferences from blood collection tube components on clinical chemistry assays. Biochem Med. 2014;24:31–44.

Google Scholar

CrossrefSearch ADS

WorldCat

 6

International Standards Organization (ISO). ISO 15189:2012(en). https://www.iso.org/obp/ui/#iso:std:iso:15189:ed-3:v2:en. Accessed July 20, 2015.

7

Clinical and Laboratory Standards Institute (CLSI). Tubes and Additives for Venous Blood Specimen Collection; Approved Standard—Sixth Edition. Document no. GP39A6E. Wayne, PA: Clinical Laboratory Standards Institute; 2010.

Google Scholar

Google Preview

OpenURL Placeholder Text

WorldCat

COPAC8

International Council for Standardization in Haematology. Recommendations of the International Council for Standardization in Haematology for ethylenediaminetetraacetic acid anticoagulation of blood for blood cell counting and sizing. Am J Clin Pathol. 1993;100:371-472.

CrossrefSearch ADSPubMed

WorldCat

 9

Banfi G, Salvagno GL, Lippi G. The role of ethylenediamine tetraacetic acid (EDTA) as in vitro anticoagulant for diagnostic purposes. Clin Chem Lab Med. 2007;45:565–576.

Google Scholar

CrossrefSearch ADSPubMed

WorldCat

 10

McKenzie SB, ed. Clinical Laboratory Hematology. 1st ed. New Jersey: Prentice Hall; 2006.

Google Scholar

Google Preview

OpenURL Placeholder Text

WorldCat

COPAC 11

CLSI. Evaluation of Precision Performance of Quantitative Measurement Methods; Approved GuidelineSecond Edition. CLSI EP5-A2 document. Wayne, PA: Clinicaland Laboratory Standards Institute; 2004.

OpenURL Placeholder Text

WorldCat

12

Mailankot M, Thushara T, Praveena P, Jacob J, Jobin Rajan B, Vasudevan DM Various Anticoagulants and Fluoride do not Affect HbA1C Level. Indian J Clin Biochem. 2012;27:209.

Google Scholar

CrossrefSearch ADSPubMed

WorldCat

 13

Chakraborty S, Ghosh SS, Das A, Sawant P, Kallner A. Can EDTA, EDTA-fluoride, and buffered citrate tubes be used for measurement of HbA1c on the Bio-Rad D10? Clin Chem Lab Med. 2015;53:5-8.

Google Scholar

CrossrefSearch ADSPubMed

WorldCat

 14

Little RR, Roberts WL. A review of variant hemoglobins interfering with hemoglobin A1c measurement. J Diabetes Sci Technol. 2009;3:446-51.

Google Scholar

CrossrefSearch ADSPubMed

WorldCat

 15

Architect System HbA1C Package insert instructions. REF 4P72, ABBL343/R05. Abbott Park, Il: Abbott Laboratories; 2013.

Abbreviations

    Abbreviations 
  • HbA1c

    glycosylated hemoglobin, type A1C

    •  
  • EDTA

    ethylenediaminetetraacetic acid

    •  
  • ICSH

    International Council for Standardization in Hematology

    •  
  • CLSI

    Clinical and Laboratory Standards Institute

    •  
  • CV

    coefficient of variation

    •  
  • Hb

    hemoglobin

    •  
  • CI

    confidence interval

© American Society for Clinical Pathology, 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com Topic:
  • anticoagulants
  • edetic acid
  • hemoglobin a, glycosylated
  • laboratory
  • synthetic cannabinoids
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