HPLC--how necessary is it for haemoglobinopathy diagnosis in India?

ABSTRACT

Cation exchange high performance liquid chromatography (HPLC) is emerging as the method of choice for the initial screening of thalassemias and haemoglobinopathies and quantification of Haemoglobins (Hbs) like HbA, HbA2 and HbF. Since it is expensive, the present study was conducted to evaluate the need for HPLC in Indian laboratories and identify situations where it would be imperative. Eighty three patients suspected to have thalassemia and haemoglobinopathies were analysed. Both HPLC and alkaline gel electrophoresis detected 14 cases of HbE syndrome and 14 cases of HbS syndrome. However of the 14 cases diagnosed as HbD syndrome by alkaline electrophoresis, eight cases were diagnosed as Hb Q India, 1 case as HbD Iran and 5 cases of HbD Punjab on HPLC. Thirty-one cases were detected to have beta heterozygous thalassemia based on the high HbA2 levels (>3.9%) and eight cases were diagnosed as beta homozygous thalassemia by both HPLC and gel electrophoresis. One of them had an unknown Hb migrating in F-A region. Her mother also had same unknown Hb variant. In view of electrophoretic migration and retention time (RT) on HPLC, possibility of HbG-San Jose was considered. HPLC being an automated instrument is highly sensitive and specific, has high resolution and helps in quantification of various haemoglobins. However in a developing country like India where economical factors play a major role in planning for management of patients, the role of HPLC is limited.