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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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IMPACT FACTOR 2017: 3.556

CiteScore 2017: 2.34

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1437-4331
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Volume 51, Issue 4

Issues

Identification and quantification of hemoglobins in whole blood: the analytical and organizational aspects of Capillarys 2 Flex Piercing compared with agarose electrophoresis and HPLC methods

Sara Altinier / Mariacristina Varagnolo / Martina Zaninotto / Mario Plebani
Published Online: 2012-10-12 | DOI: https://doi.org/10.1515/cclm-2012-0061

Abstract

Background: The present study was conducted to evaluate the analytical performance and the organizational aspects of Capillarys 2 Flex Piercing system (CFP) respect to agarose electrophoresis and HPLC methods in hemoglobinopathies screening.

Methods: The measurement of imprecision in HbA2 and HbF quantification was verified on HbA2 CFP control and on three samples; 74 whole blood samples were used to evaluate migration time imprecision of hemoglobin variants S, C and E (HbS, HbC, and HbE); to compare methods, 451 samples were tested on CFP and HPLC; reference values were verified as value distribution in 160 blood donors and at ROC curve analysis on 449 samples from routine analysis.

Results: Imprecision: the analytical CV%s ranged from 1.25 to 3.9 at HbA2 quantification, the CV% was 3.78 at HbF quantification; the running time imprecision for HbS and HbC and HbE ranged from 0.20 to 0.69%. Method comparison: at regression analysis findings were HbA2: CFP=1.21×HPLC–0.64, HbF: CFP=1.31×HPLC-0.75, HbS: CFP=1.10×HPLC-3.24. Reference values: the HbA2 95th percentile range was 2.5–2.8; HbF was undetectable in 154 out 160 samples tested; at ROC curve analysis the best combination of sensitivity and diagnostic efficiency was obtained using 2.2 and 3.0, as reference values, for HbA2 and 1.1 as the upper reference limit for HbF. Organizational aspects: with respect to the procedures currently implemented in our laboratory CFP requires 2 h less time and obviates the need for some manual steps.

Conclusions: The quantification, reproducibility and diagnostic efficiency provided by CFP in identification and quantification of hemoglobins appear accurate. In addition, the use of primary tubes allows improved safety, and the avoidance of some manual steps, that prolong working time and are a source of possible errors.

Keywords: capillary electrophoresis; hemoglobins; HPLC

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About the article

Corresponding author: Sara Altinier, Department of Laboratory Medicine, University-Hospital of Padova, Via Giustiniani 2 35128 Padova, Italy, Phone: +39 049 821 8708, Fax: +39 049 8218489


Received: 2012-01-31

Accepted: 2012-09-06

Published Online: 2012-10-12

Published in Print: 2013-04-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 51, Issue 4, Pages 791–797, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2012-0061.

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