Main Article Content
Objective: The study aimed to evaluate the accuracy of hemoglobin analysis by the Capillarys 2 Flex Piercing using capillary electrophoresis (CE) method compared with the Variant II using high pressure liquid chromatography (HPLC) for reference method.
Material and Method: A total of 344 routine EDTA blood samples were collected to diagnose thalassemia and hemoglobinopathies.
Results: Regarding normal Hb typing, carrier, thalassemia and hemoglobinopathies as the percentage of HbA2, Hb E and Hb F detected by Capillarys 2 Flex Piercing (CE) and the reference method (HPLC) did not significantly differ (p > 0.05), i.e., 337 samples agreed with the hemoglobin typing interpretation and 7 samples disagreed (6 of the Capillarys 2 Flex Piercing reported CSA2A, a suspected Hb CS trait). However, Variant II reported A2A and 1 of the Hb type = A2EA from Capillarys 2 Flex Piercing but Variant II reported A2A (A2 = 3.8%) indicating the β-thal trait. Using the coefficient of variation the within-run and between-run precision was less than 5%
Conclusion: The Capillarys 2 Flex Piercing is capable of identifying and quantifying hemoglobin species. The major advantage of the CE method is its ability to separate and quantitate HbA2, Hb E which are important parameter required to diagnose β-thal trait. Alternatively a blood transfusion could successfully detect unstable Hb as Hb CS in the heterozygous state. This instrument proved useful in providing relevant data to investigate thalassemia and hemoglobinopathies.
2. Clarke GM, Higgins TN. Laboratory investigation of haemoglobinopathies and thalassemia: review and update. Clin Chem.2000;46:1284-90.
3. Winichagoon P, Svasti S, Munkongdee T, Chaiya W, Boonmongkol P, Chantrakul N, et al. Rapid diagnosis of thalassemiaa and other hemoglobonopathies by capillary electrophoresis system. Transl Res. 2008;152:178-84.
4. Wangrungrot P, Changsri K. Deviation of abnormalhemoglobin diagnosis using hemoglobin typing technic. J Hematol Transfus Med. 2014;24:151-6.
5. Agouti I, Merono F, Bonello-Palot N, Badens C. Analytical evaluation of the Capillarys 2 Flex piercing for routine haemoglobinopathies diagnosis. Int J Lab Hematol. 2013;35:217-21.
6. Sangkitporn S, Sangkitporn SK, Tanjatham S, SuwannakanB, Rithapirom S, Yodtup C, et al. Multicenter validation of fully automated capillary electrophoresis method for diagnosis of thalassemias and haemoglobinopathies in Thailand. Southeast Asian J Trop Med Public Health. 2011;42:1224-32.
7. Borbely N, Phelan L, Szydlo R, Bain B. Capillary zone electrophoresis for haemoglobinopathy diagnosis. J Clin Pathol. 2013;66:29-39.
8. Altinier S, Varagnolo M, Zaninotto M, Plebani M. 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. Clin Chem Lab Med. 2012;12:1-7.
9. Higgins TN, Khajuria A, Mack M. Quantification of Hb A2 in patients with and without β- thalassemia and in the presence of Hb S, Hb C, Hb E, and Hb D Punjab hemoglobin variants. Am J Clin Pathol. 2009;131:357-62.
10. Greene DN, Pyle AL, Chang JS, Hoke C, Lorey T. Comparison of Sebia Capillarys Flex capillary electrophoresis with the BioRad Variant II high pressure liquid chromatography in the evaluation of hemoglobinopathies. Clin Chim Acta. 2012;16:1232-8.
11. Boonkant P, Sangkitporn S, Yodtup C, Masasso S, Sangkitporn S. Validation of automated hemoglobin analyzer: Capillarys 2 Flex Piercing for diagnosis of thalassemias and hemoglobinopathies. J Med Assoc Thai. 2013;41:4457-71.
12. Sysmex Corporation. Automated hematology analyzer XT-2000i/XT1800i instructions for use. Kobe: The Corporation; 2010.
13. Lab manual of thalassemia and hemoglobinopathies investigation. Revised edition. Clinical Research Center Department of Medical Sciences. Ministry of Public Health. 2015.
14. Westgard JO, Carey RN, Wold S. Criteria for judging precision and accuracy in methog development and evaluation. Clin Chem. 1974;20: 825-33.
15. Waneesorn J, Panyasai S,Kongthai K, SingboottraP, Pornprasert S. Comparison between capillary electrophoresis and high performance liquid chromatography for detection and quantification of Hb Constant Spring. Hemoglobin. 2011;35:338-45.
16. Pornprasert S, Panyasai S, Waneesorn J, Kongthai K, Singboottra P. Quantification of hemoglobin Constant Spring in heterozygote and homozygote by a capillary electrophoresis method. Int J Lab Hematol. 2012;34:143-7.