Usefulness of High Performance Liquid Chromatography and Red Cell Indices as a Screening Tool for Diagnosing Haemoglobinopathies: A Retrospective Observational Study from North India

Introduction: The World Health Organization estimates that seven percent of the global population is a carrier for disorders of haemoglobin with Thalassemia and haemoglobinopathies being the commonest genetic disorders of haemoglobin. As an initial screening method, Red Blood Cell indices like total count of Red Blood Cell (RBC), Mean Cell Volume (MCV), Mean Cell Haemoglobin (MCH), Mean Corpuscular haemoglobin concentration (MCHC) and Red Cell Distribution Width (RDW) can be utilized. High Performance Liquid Chromatography (HPLC) has surfaced as a powerful, excellent screening tool for direct identification of different haemoglobin variants.

Aim: This study aims at using HPLC to identify different haemoglobinopathies and find out the distribution of different red blood cell (RBC) indices in different haemoglobinopathies to determine their significance as screening test.

Materials and Methods: A retrospective study was carried out on 73,333 cases from January 2017 to October 2022 (5 years). EDTA samples were run on automated haematology analyser for red blood cell indices before doing HPLC and then analysed on the Bio-Rad Variant II CE-HPLC system with use of the Variant II-Thalassemia short program (Bio-Rad Laboratories) for determination of different fractions of haemoglobin level.

Results: 6242 (8.52%) cases of abnormal haemoglobin variants were recorded. The most prevalent haemoglobin (Hb) fraction was Beta Thalassemia trait (5.59%) followed by HbD Punjab Heterozygous, HbE Heterozygous and Sickle cell heterozygous. RBC indices were statistically significant between subjects with normal haemoglobin study and those with haemoglobipathies (p<0.0001). RBC indices were also statistically significant between beta Thalassemia trait and subjects with normal haemoglobin study (p<0.0001). Mean value of RDW is markedly increased in both Thalassemia syndrome (36.06±6.79) and Hb-E Beta Thalassemia (32.49±6.57). Mean total RBC count was highest in Beta Thalassemia trait (5.07±0.90) and lowest in Beta Thalassemia syndrome (2.71±1.10).

Conclusion: RBC indices can be utilized to screen Thalassemia and other haemoglobinopathies and used as a supportive test to High Performance Liquid Chromatography (HPLC)