–Thalassemia is most common inherited red cell disorder worldwide and single gene disorders in India . Effective population screening of β thalassemia trait can dramatically decrease the incidence of birth of a thalassemia major child. Through genetic counseling birth rate of β–thalassemia major can be reduced by 90%.Aim of the study was to determine the prevalence of –Thalassemia in patients with microcytic hypochromic anemia and to assess the suitability of using CBC, PBF, NESTROFT and confirmation by HPLC. A total of 666 cases from January 2016 to December 2018 of microcytic hypochromic anaemia submitted for HPLC in the Department of Pathology, Jawahar Lal Nehru Medical College, Ajmer. Out of 666 microcytic hypochromic cases, 12 cases were diagnosed as –thalassemia major ( BTM), 106 cases –thalasemia trait ( BTT) and rest were of non – thalassemia micocytic hypochromic(NBT) anemia.. Majority of –thalassemia (37.28% ) were seen in the paediatric age group of 0–15 years. BTM was more common in males than females. Majority of the cases were Hindus (66.94%) followed by Muslims(16.94%) and Sindhi population (14.40 %). Mean RBC count , Hb g/dl, and Hct were significantly higher and MCV , MCH and RDW were significantly lower in BTT . Mentzers index and Green and Kings index were highly reliable with high specificity and sensitivity along with NESTROFT(Naked Eye Single–Tube Red Cell Osmotic Fragility Test). A set of screening tests like NESTROFT, Mentzers index and Green and Kings index along with routine hemogram data (RBCcount ,Hb,MCV,MCH,RDW ) in microcytic hypochromic cases can effectively discriminate between BTT and Non BTT, and diagnosis of BTT can be reliably done by HPLC and HbA2 quantitation by elution with HbA2 >3.5%.