Original Article

Hematological indices and hemoglobin high-pressure liquid chromatography for diagnosis of hemoglobinopathies in a secondary care center in North-East India

Mahendra Narain Mishra1, Bianchi Sangma2, Divya Mary George2, Lydia John3, Koshy C. George2

1Department of Pathology, Baptist Christian Hospital, Tezpur, Ketkibari, Mission Charalie, Assam, India
2Department of Pediatrics, Baptist Christian Hospital, Tezpur, Ketkibari, Mission Charalie, Assam, India
3Department of Internal Medicine, Baptist Christian Hospital, Tezpur, Ketkibari, Mission Charalie, Assam, India


Background: Hemoglobinopathies occur globally and are often presented with anemia. This study was performed in a secondary care center in Sonitpur District, Assam, to study the usefulness of hematological indices and hemoglobin High-Pressure Liquid Chromatography (HPLC) for characterization of hemoglobinopathies and to quantify the prevalence and types of anemia in a resource- poor setting of North-East India.
Methods: Data of 9936 hemoglobin estimations and 708 peripheral blood smear examinations performed over a year were retrieved. Complete blood count for 170 patients was performed by XS 800i Five-Part Sysmex Cell Counter whereas hemoglobin (Hb) HPLC was outsourced. Serum iron estimation was done for 100 samples by dry chemistry and serum ferritin assay was tested by direct chemiluminescence for 13 patients. The number of hospital visits, hospitalization duration, blood transfusions, demographic profile, and unusual features in some patients was recorded.
Results: Anemia (Hb < 11.0 g/dl) was present in 79.6% of 9936 samples tested. Microcytic hypochromic anemia was present in 278/534 (52%) patients. The mean age of 170 subjects was 15.4 years (3 months ? 56 years) with a slight female preponderance 87/51.2%. Erythrocytosis was observed in 32/18.8% of samples, of which 18 were females. Microcytosis and low mean corpuscular hemoglobin (MCH) were observed in 143 (84.1%) and 153 (90%) samples, respectively. The number and percentage of conditions identified by HPLC are no abnormality in 53 (31.2%), hemoglobin E disease 31 (18.2%), hemoglobin E trait 25 (14.7%), ?-thalassemia minor 11 (6.5%), ?-thalassemia major 4 (2.4%), compound hemoglobinopathy 15 (8.8%), sickle cell trait 12 (7%) and sickle cell disease 8 (4.7%), and inconclusive in 11 (6.5 %) patients. Serum iron was low in 39 (34.5%), normal in 65 (57.5%), and high in 9 (8%) of the 113 subjects tested.
Conclusions: Prevalence of anemia and hemoglobin E abnormality was high with unexpected severe anemia in some heterozygotes for HbE, HbS, and ?- thalassemia. Hemoglobin HPLC was useful in arriving at a presumptive diagnosis and must be used as a frontline investigation even in resource-poor settings

Keywords: anemia; chromatography; hemoglobinopathies; India; iron