Perspectives In Medical Research (Dec 2017)
Evaluation of (HPLC) Patterns of Sickle Cell Anaemia patients in comparison with apparently healthy individuals
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: 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 and to study the Hb chromatographic patterns according to the gender, age and blood groups. Also to evaluate the efficiency of variant Hb testing system in detection of HbS type of sickle cell anaemia patients. Materials & Methods: A total of Thirty five sickle cell anemia patients who were attending to the Prathima Institute of Medical Sciences ,Teaching Hospital, Karimnagar Telangana state, the samples were (25) males and (10) females , from February of 2017 to April of 2017 . And Fifteen 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 these paration and quantification of Hb types by high performance liquid chromatography technique. Results: The study of Hb chromatographic patterns of samples revealed that there were highly significant differences (P= 0.001) for HbA2,HbA,HbFand 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 that the mean of the highest HbA and HbA2 percentages of patients were (8.13±7.09 and 2.82±1.32) respectively in age group (6-10) years ,HbF (17.32±6.63) in 11- 15yrs age group,while it was (74.55±2.35) for HbS type in >16yrs age group. As for control group, the highest HbF percentages was (0.78±0.18) in age group less than five years old, for HbA was (86.5±3.69) in age group (6-10) years old, for HbA2 was (2.13±0.32) in age group (11-15) years old, and for HbSwas(0.00) in all age groups. . 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: The study of Hb chromatographic patterns is useful for the diagnosis of sickle cell anaemia. 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. HPLC is an excellent, powerful diagnostic tool for the direct identification of HbS.
