Aim: The aim of the study is to evaluate the clinical relevance of the immature reticulocyte fraction (IRF) and reticulocyte maturity indices in distinguishing between various causes of anaemia in patients. Methods: The present study is conducted in the Department of Pathology for a period of one year and 100 cases were included in the study. Results: The result showed mean retic count of 1.3 with SD of 0.007, and in IRF mean was 0.091 with SD of 0.120. Retic count was low in IDA compared to thalassemia patient (high- hemolytic), IRF values were low in IDA and in CKD patient compared to thalassemia group and regarding reticulocyte indices, statistically significant difference was observed for RET H, RET M and RET L between three groups. Conclusion: Our study showed that with the use of automated fluorescence analyser providing IRF and reticulocyte maturity indices (LFR, MFR and HFR) was quite very useful in the early detection and for the differential diagnosis of iron deficiency anemia, thalassemia and anemia due to chronic kidney disease.