Experience with NESTROFT for screening for thalassemia trait/ minor: evaluation against CBC and HPLC in a high prevalence region in Saurashtra, Gujarat, India
Keywords:Beta thalassemia trait, Carrier screening, Complete blood count, High performance liquid chromatography, Naked eye single tube red cell osmotic fragility test
Background: Hemoglobinopathies pose a significant health burden in India. Prevention programmes can significantly reduce this burden. Although sophisticated methods of screening for β thalassemia trait are available, a cheap and simple method is beneficial for population screening. Although the Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT) has been evaluated in many studies, sample sizes were small in some and many earlier studies have not done complete blood count (CBC) and High-Performance Liquid Chromatography (HPLC) in all the cases. We evaluate the suitability of NESTROFT for detection of β-thalassemia trait in a high prevalence region in Saurashtra, Gujarat.
Methods: Here, 1000 unrelated individuals were studied. NESTROFT, CBC and estimation of HbA2 and HbF or other hemoglobin variants were done by HPLC.
Results: Prevalence of β thalassemia trait was 7.8% in this population. NESTROFT showed an overall sensitivity and specificity of 94.87 and 85.38 respectively for the detection of β thalassaemia trait. Using red cell indices (MCH <27 pg and MCV <80 fl), One β thalassemia trait with normal indices would have been missed. Among twelve individuals with other hemoglobinopathies (HbS, HbD, HbE, δβ thalassemia trait or HPFH), seven had a positive NESTROFT while three had normal MCV & MCH values.
Conclusions: NESTROFT is a cost-effective sensitive test which does not require any equipment and can be done in remote areas. It remains a useful first line screening test when large populations have to be screened.
Williams TN, Weatherall DJ. World distribution, population genetics, and health burden of the hemoglobinopathies. Cold Spring Harbor Perspec Med. 2012 Sep 1;2(9):a011692.
Madan N, Sharma S, Sood SK, Colah R, Bhatia HM. Frequency of β-thalassemia trait and other hemoglobinopathies in northern and western India. Ind J Human Gene. 2010 Jan;16(1):16.
Colah RB, Gorakshakar A. Control of thalassemia in India. Thalass Rep. 2014 Sep 29.
Colah R, Gorakshakar A, Phanasgaonkar S, D’Souza E, Nadkarni A, Surve R, et al. Epidemiology of β‐thalassaemia in Western India: mapping the frequencies and mutations in sub‐regions of Maharashtra and Gujarat. Bri J Haematol. 2010 Jun;149(5):739-47.
Patel Ashwin P, Naik Madhuben R, Shah Nilam M, Sharma Narmadeshwar P, Parmar Prakash H. Prevalence of common hemoglobinopathies in Gujarat: an analysis of a large population screening program. Natl J Commu Med. 2012;3:112-7.
Kattamis C, Efremov G, Pootrakul S. Effectiveness of one tube osmotic fragility screening in detecting beta-thalassaemia trait. J Med Gene. 1981 Aug 1;18(4):266-70.
Gorakshaker AC, Colah R, Nadkarni A, Desai S. Evaluation of the single tube osmotic fragility test in detection of ß-thalassaemia trait. Natl Med J Ind. 1990;3:171-3.
Bhukhanvala D, Seliya V, Shah A, Gupte S. Study of parents of β-thalassemia major children to determine cutoff values of hematological parameters for diagnosis of β-thalassemia trait and assessment of anemia in them. Ind J Med Sci. 2013 May 1;67.
Angastiniotis M, Modell B, Englezos P, Boulyjenkov V. Prevention and control of haemoglobinopathies. Bull World Health Organ. 1995;73(3):375.
Thomas S, Srivastava A, Jeyaseelan L, Dennison D, Chandy M. NESTROFT as a screening test for the detection of thalassaemia and common haemoglobinopathies--an evaluation against a high performance liquid chromatographic method. Ind J Med Res. 1996 Aug;104:194-7.
Gomber S, Madan N. Validity of nestroft in screening and diagnosis of β-thalassemia trait. J Trop Pediatr. 1997 Dec 1;43(6):363-6.
Bobhate SK, Gaikwad ST, Bhaledrao T. NESTROFF as a screening test for detection of Beta-thalassemia trait. Ind J Pathol Microbiol. 2002 Jul;45(3):265-7.
Chow J, Phelan L, Bain BJ. Evaluation of single‐tube osmotic fragility as a screening test for thalassemia. Am J Hematol. 2005 Jul;79(3):198-201.
Singh SP, Gupta SC. Effectiveness of red cell osmotic fragility test with varying degrees of saline concentration in detecting beta-thalassaemia trait. Singapore Med J. 2008 Oct 1;49(10):823.
Sumera A, Ahmed S, Ali SA, Khanani R. Evaluation of NESTROFT as a marker of differentiation between β-thalassemia trait and iron deficiency anemia. International Journal of Collaborative Res Inter Med Pub Health. 2012;4(8): 1560-66.
Detection by NESTROFT: An answer in rural scenario? Iranian J of Pathol. 2012;7:19-26.
Mehta BC, Gandhi S, Mehta JB, Kamath P. Naked eye single tube red cell osmotic fragility test for beta-thalassaemia: population survey. Ind J Haematol. 1988;6:187-90.
Mendiratta SL, Bajaj S, Popli S, Singh S. Screening of women in the antenatal period for thalassemia carrier status: comparison of NESTROFT, red cell indices, and HPLC analysis. J Fetal Med. 2015 Mar 1;2(1):21-5.
Ismail M, Patel NG. Effectiveness of naked eye single tube osmotic fragility test for screening of β-thalassemia trait from north Maharashtra region, India. Inter J Commu Med Pub Health. 2016;3:1255-60.
Sharma G, Sharma D, Gulati RK. Use of NESTROFT as a screening test for the carriers of thalassemia major. InterJ Pediatr Res. 2016;3:502-6.