Evaluation of (HPLC) Patterns of Sickle Cell Anaemia Patients in Comparison with Apparently Healthy Individuals

Mohammad Ali Jeddoa, Zuhair

	Evaluation of (HPLC) Patterns of Sickle Cell Anaemia Patients in Comparison with Apparently Healthy Individuals
Kerbala Journal of Medicine
Article 9, Volume 4, Issue 9, 2011, Pages 980-987 PDF (0 K)
Author
Zuhair Mohammad Ali Jeddoa
Abstract
background: Sickle cell haemoglobin (HbS) results from an autosomal recessively inherited mutation in which the amino-acid glutamine is replaced by valine at position 6 in the beta globin chain of haemoglobin (Hb). Sickle cells have a reduced deformability and are easily destroyed, causing occlusion of the microcirculation and a chronic haemolytic anaemia with a median Hb concentration level of about 9 g/dl. Routine electrophoresis methods and High performance liquid chromatography (HPLC) were used to screen normal and variant Hb, and allowed the verification of the Hb observed with electrophoresis and precise quantification of their proportion. Objectives: 1- This study aimed to evaluate the chromatographic pattern of Hb types (HbA, HbF, HbA2 and HbS) of sickle cell anemia patients in comparison with the apparently healthy individuals. 2- To study the Hb chromatographic patterns according to the gender, age and blood groups. 3- To evaluate the efficiency of variant Hb testing system in detection of HbS type of sickle cell anaemia patients. Methods: A total of eighty four sickle cell anemia patients who were attending to the Ibn Al-Baladi pediatrics hospital (in Baghdad) and Kerbala teaching pediatrics hospital, the samples were (39) males and (45) females , from April of 2006 to February of 2007 . And thirty seven of case controls with matched age and sex were randomly selected from apparently healthy individuals. High performance liquid chromatography (HPLC) was adopted to determine the different types of Hb for patients and control groups using variant Hb testing system which depend upon the separation and quantification of Hb types by high performance liquid chromatography technique. Results: The study of Hb chromatographic patterns of samples revealed that there were no significant differences (p ≥ 0.05) between the values of HbA2 for the patients and control groups and there were significant differences (P≤ 0.05) for HbF and highly significant differences (P≤ 0.01) for HbA and HbS percentages of patients in comparison with the control group. The results of Hb chromatographic patterns of samples according to the gender revealed that there were no significant differences at (p≥ 0.05) between males and females within patient and control groups. The results of Hb chromatographic patterns of Hb types for patients and control groups according to the age groups revealed heterogeneous results with highest HbF, HbA, and HbA2 percentages of patients were (10.83±3.32, 57.6±7.33, and 4.22 ±1.88) respectively in age group less than five years old, while it was (54.33±8.9) for HbS type in age group (6-10) years old. As for control group, the highest HbF percentages was (8.2±4.09) in age group less than five years old, for HbA was (87.22±5.86) in age group (6-10) years old, for HbA2 was (3.6±0.23) in age group (11-15) years old, and for HbS was (0.3±0.05) in age group less than five years old. Finally, the results also showed that there were no significant differences at (P ≥ 0.05) for Hb chromatographic patterns of different Hb types percentages according to the blood groups of studied samples within group (patients or controls). Conclusion: 1- The study of Hb chromatographic patterns is useful for the diagnosis of sickle cell anaemia. 2- There are no significant effects of gender and blood groups on the chromatographic patterns of different Hb types of sickle cell anaemia patients in comparison to the apparently healthy individuals. 3- HPLC is an excellent, powerful diagnostic tool for the direct identification of HbS.
Keywords
Iraq; Sickle cell anemia; HbS; HPLC; Variant Hb testing system

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