Document Type : Original Research


1 Dept. of Pathology, North Bengal Medical College, Siliguri, India

2 Dept. of Pathology, Medical College, Kolkata, India

3 Dept. of Community Medicine, West Bengal University of Health Sciences, Siliguri, India


Background and Objectives: Beta-thalassemia continues to be a cause of significant burden to the society particularly in the poorer developing countries. Although sophisticated methods of screening have become available, a hunt for a cheap, rapid, objective screening method still remains elusive. Thus, the objectives are to study the validity of Naked-Eye-Single-Tube-Osmotic-Fragility-Test (NESTROFT) in detection of beta-thalassemia carrier state, to assess the prevalence of beta-thalassemia trait among antenatal mothers in the region and also to find out the effect of concurrent iron deficiency on the hematological parameters in these cases.
Materials and Methods:A total of 500 antenatal mothers in a rural tertiary care hospital were selected for the study. Their blood samples were subjected to NESTROFT, complete hemogram, reticulocyte counts and hemoglobin variant studies by electrophoresis and by high pressure liquid chromatography (HPLC). Serum ferritin estimation was done in cases diagnosed as beta-thalassemia trait. The results were analyzed statistically.
Results: A prevalence of 3.4% of beta-thalassemia trait and 0.6% of E-beta-thalassemia were observed among the study population. NESTROFT showed an overall sensitivity and specificity of 95% and 95.8% respectively in detection of heterozygous and double heterozygous states of beta-thalassemia. The various RBC indices were significantly (P <0.05) lower in carriers with concurrent iron deficiency. A co-existent iron deficiency did not preclude a diagnosis of beta-thalassemia carrier state.
Conclusions: NESTROFT appears to a valid test in rural setting with financial constraints. The hematological parameters in iron deficient beta-thalassemic carriers significantly differed from their iron replete counterparts but did not cause problem in diagnoses.


  1. Sachdev R, Dam AR, Tyagi G. Detection of Hb variants and hemoglobinopathies in Indian population using HPLC: report of 2600 cases. Indian J Pathol Microbiol 2010 Jan;53(1):57-62.
  2. Galanello R, Eleftheriou A, Trager-Synodinos J, Old J, Petrou M, Angastiniotis M. Prevention of thalassaemias and other haemoglobin Disorders. Nicosia: Thalassaemia International Federation Publication; 2003.
  3. Choudhury V, Kotwal J, Saxena R. Thalassemia screening and control programme. Pediatrics Today 1998;(1):283-9.
  4. Verma IC, Saxena R, Thomas E, Jain PK. Regional distribution of beta-thalassemia mutations in India. Hum Genet 1997 Jul;100(1):109-13.
  5. Gajra B, Chakraborti S, Sengupta B. Prenatal Diagnosis of Thalassaemias. Int J Hum Genet 2002 May;2(3):173-8.
  6. Piplani S. Hemoglobin E disorders in the north east India. J Assoc Physicians India 2000 Nov;48(11):1082-4.
  7. Mamtani M, Jawahirani A, Das K, Rughwani V, Kulkarni H. Bias-corrected diagnostic performance of the naked-eye single-tube red-cell osmotic fragility test (NESTROFT): an effective screening tool for beta-thalassemia. Hematology 2006 Aug;11(4):277-86.
  8. Desaei M, Desaei P. Do mothers with hemoglobinopathies and anemia require iron supplementation in pregnancy? J Obs Gynaec India 1994 Jun;(44):177-80.
  9. Sinha M, Panigrahi I, Shukla J, Khanna A, Saxena R. Spectrum of anemia in pregnant Indian women and importance of antenatal screening. Indian J Pathol Microbiol 2006 Jul;49(3):373-5.
  10. Sur D, Mukhopadhyay SP. Prevalence of thalassaemia trait in the state of West Bengal. J Indian Med Assoc 2006 Jan;104(1):11-5.
  11. Thomas S, Srivastava A, Jeyaseelan L, Dennison D, Chandy M. NESTROFT as a screening test for the detection of thalassaemia & common haemoglobinopathies--an evaluation against a high performance liquid chromatographic method. Indian J Med Res 1996 Aug;104:194-7.:194-7.
  12. Maheshwari M, Arora S, Kabra M, Menon PS. Carrier screening and prenatal diagnosis of beta-thalassemia. Indian Pediatr 1999 Nov;36(11):1119-25.
  13. Suri V, Sidhu P. Evaluation of N.E.S.T.R.O.F.T. as a screening test in detection of â-thalassemia trait. Int J Hematology and Blood transfusion 2001 Jan;(19):6-7.
  14. Bobhate SK, Gaikwad ST, Bhaledrao T. NESTROFF as a screening test for detection of Beta-thalassemia trait. Indian J Pathol Microbiol 2002 Jul;45(3):265-7.
  15. Sirichotiyakul S, Tantipalakorn C, Sanguansermsri T, Wanapirak C, Tongsong T. Erythrocyte osmotic fragility test for screening of alpha-thalassemia-1 and beta-thalassemia trait in pregnancy. Int J Gynaecol Obstet 2004 Sep;86(3):347-50.
  16. Mamtani M, Das K, Jawahirani A, Rughwani V, Kulkarni H. Is NESTROFT sufficient for mass screening for beta-thalassaemia trait? J Med Screen 2007;14(4):169-73.
  17. Madan N, Sikka M, Sharma S, Rusia U. Haematological parameters and HbA2 levels in beta-thalassaemia trait with coincident iron deficiency. Indian J Pathol Microbiol 1998 Jul;41(3):309-13.