The evaluation of capillary electrophoresis (CE) performance and comparison with high performance liquid chromatography (HPLC) system for the diagnosis of thalassaemia and haemoglobinopathies

Lai Hung (Edith) Woo, Michael A. Cahill, Azizur Rahman, Chun Sing (Stephen) Chow

Research output: ThesisDoctoral Thesis

Abstract

Capillary electrophoresis (CE) can be used as an initial screening method for patients with haemoglobinopathies. The aim of this study is to compare two methodologies to identify haemoglobinopathies, i.e., high pressure liquid chromatography (HPLC) and CE, and to suggest an effective workflow for the screening of haemoglobinopathies in the laboratory. A total of 576 blood samples were analyzed in this study for comparison, of which 67 samples were from diabetic patients with abnormal HPLC chromatograms. Both HPLC and CE analysis were performed. The normal level of haemoglobin (Hb) A2 (HbA2) was measured in 215 blood samples by both CE procedure and HPLC with no statistical difference with p-value of 0.1488. The level of HbF was measured by CE method in 118 blood samples. Normal and elevated levels of HbF were in agreement with the comparative HPLC with p-value of 0.6311. About 91 different blood samples with haemoglobin variants, such as HbS, HbE, HbJ, HbD/G and HbQ-Thailand were analyzed with CE and HPLC. All of them were statistically significant in different detected levels of haemoglobin variants (p <0.05). All abnormal haemoglobin variants were detected and there were no cases of false positive observed by comparison with the HPLC system. The CE system proved to be suitable for the identification of major Hb variants as well as detection of abnormal haemoglobins, thereby providing useful information for presumptive diagnoses. CE also can be used to quantify HbH and Hb Bart’s in HbH disease, alpha thalassaemia in adult and newborn patients. Particularly, CE advantageously measures HbA2 in the presence of HbE, whereas HPLC was unable to differentiate beta- thalassaemia from benign conditions because HbE and HbA2 co-eluted with the same retention time by HPLC.
Original languageEnglish
QualificationMaster of Science
Awarding Institution
  • Charles Sturt University
Supervisors/Advisors
  • Cahill, Mike, Principal Supervisor
  • Rahman, Azizur, Co-Supervisor
  • Chow, Chun Sing (Stephen), Co-Supervisor, External person
Award date17 Dec 2014
Publication statusPublished - 2014

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