DOI: https://doi.org/10.53350/pjmhs221651225 ORIGINAL ARTICLE P J M H S Vol. 16, No. 05, May 2022 1225 Evaluation of Srivastava index to distinguishing Beta-Thalassemia Trait from Iron Deficiency HASSNAIN SHAHID 1 , MARYAM SALEEM 2 , NAUMAN NASEER 3 , SAMINA TABUSSAM 4 , ATIF AZIZ 5 , SAEED ULLAH 6 1 Resident Paeds Pak Emirates Military Hospital (PEMH) Rawalpindi 2 House officer Pak Emirates Military Hospital (PEMH) Rawalpindi 3 Classified Child Specialist Fellow neonatologist CMH Rawalpindi 4 Assistant Professor Pediatrics CMH Institute of Medical Sciences Bahawalpur 5 CMH Peshawar 6 Resident paeds Pak Emirates Military Hospital (PEMH) Rawalpindi Corresponding author: Hassnain Shahid, Email: husnain_1115@hotmail.com, Phone: +923325568986 ABSTRACT Objective: To assist in the differential diagnosis of beta thalassemia trait and iron deficiency anaemia, many alternative red blood cell index-based formulae were examined. Methods: The Rawalpindi PEMH performed this study from June 2021 to March 2022. For individuals with beta- thalassemia trait and iron deficient anaemia, age and gender were not considered. More than five millilitres of blood were drawn from each patient in order to determine the haemoglobin content, the number of red blood cells, how they were distributed, and the average cell volume. Five alternative formulas may be used to differentiate these two circumstances. Shine and Lal index, Mentzer index, Srivastava index and Green & King index are among the most well- known ones. Youden's index was included in the calculation of sensitivity, specificity, and positive and negative predictive values (PPVs). Results: The iron deficiency anaemia rate was 70%, and the beta thalassemia phenotype was 30% among the 1500 participants. This indicator has a sensitivity of 100% and specificity of 93.3% when it comes to discriminating between beta- thalassemia trait and iron deficiency anaemia. Conclusion: The red cell distribution width index may be used to differentiate between beta thalassemia trait and iron deficiency anaemia. If a haemoglobin electrophoresis is not available, a diagnosis of beta thalassemia may still be made. Keywords: Srivastava index, beta-thalassemia, Anemia, Cell redness, Hb electrophoresis, INTRODUCTION It is the most prevalent cause of hypochromic and microcytic anaemia, beta-thalassemia trait (BTT). 1 . In a WHO study, more than half of all instances of anaemia are caused by iron deficiency (IDA). Anemia is a major cause of death and disease in developing and poor countries. Untreated, it may cause developmental delays in children, a decline in IQ, an increase in mortality, and even cause death in certain cases 2 . Pregnant women who have several pregnancies and poor health are more likely to suffer from iron deficiency anaemia (IDA) 3 . Haemoglobin production of the normal globin peptide chains is hindered in thalassemic haemoglobinopathy, a hereditary hemoglobinopathy, because of this impairment (Hb). It's the most important differential while evaluating a child with anemia 4 . The form of thalassemia observed varies by race, however it is a common genetic condition with population -dependent frequency and severity. One in five people on Earth has Hb production faults, and seven percent are Hb disease carriers 5 . 80 percent of the infants born each year with severe haemoglobinopathies are from poor or impoverished nations (annually 0.3-0.5 million) 6 . The thalassemia gene is present in a small percentage of the populations of the Eastern Mediterranean, Southeast Asia, and Sub-Saharan Africa 7 . 8 People from thalassemia-endemic countries have migrated to North America and Europe, where the prevalence of thalassemia is now much higher than previously anticipated 8 . The Middle East is responsible for 10% of all Thalassemia cases in the globe, while Southeast Asia and the Mediterranean account for 9% and 8%, respectively 9 . As the most common genetic disease in Pakistan, around 115,000 new instances of homozygote thalassemia are diagnosed each year. Pakistan; around 5% of Pakistanis are heterozygous for beta thalassemia 10 . Analyzing iron and ferritin using electrophoresis as well as haemoglobin alpha 2 (HbA2). For the BTT diagnosis, it is possible to assess HbA2 levels using cellulose acetate, gel electrophoresis, and high-performance liquid chromatography 11,12 . BTT and IDA may be distinguished using a variety of electronic haematological cell counter red cell indices. Indi ces are used to differentiate between BTT and IDA in order to save both time and money. It may be difficult to distinguish between BTT and IDA without using red cell indices 13,14 . The MCV, MCH, MCHC, and RDW indices and factors, as well as the red cell distribution width, may be used to discriminate between BTT and IDA (RDW). Finding the most BTT patients while simultaneously removing those with IDA as a factor to consider when determining the optimal discrimination index (high specificity). In this study, five indices-based formulae were tested for their sensitivity and specificity 15,16 . MATERIAL AND METHODS Pakistan's emirate military hospital in Rawalpindi (PEMH) performed this study between June 2021 and March 2022. Prior to blood samples being obtained, patients and their parents were given the option to provide their informed consent. The study's subject group included patients with microcytic hypochromic anaemia who were sent to the PEMH pathology lab for Hb electrophoresis. The research included both male and female patients between the ages of 1 and 12. Patients who had had blood transfusions within the previous four weeks or who were in a life-threatening condition were ruled out of the trial. Blood samples were collected through aseptic EDTA and plain gel tubes are used for biochemical and haematological analysis after venipuncture. Blood iron levels were measured in gel tubes after 5 minutes of spinning at 2800 RPM with Merck kits on Rayto (RT- 9200) chemical analyzers. Analyses of blood parameters such as haemoglobin (Hb), RBC count, and RBC indices such as MCV and MCHC were carried out using an automated haematology analyzer from Tokyo (Nihon Kohden). An alkaline technique is required to estimate HbA2 levels using BTT. It was discovered through the use of Cellulose Acetate Electrophoresis (Wealtech, United States). SPSS 22 was used for the statistical analysis. It was determined that several BTT indices had a high screening efficiency using the ROC curve. Several previously suggested methods were used to estimate the findings' sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) (NPV). 22,23 RESULTS We generated and tallied the five discrimination indices that were utilized in the assessment (Table-1). 1050 of the 1500 blood