APPLICATION OF INFRARED SPECTROSCOPY FOR CLASSIFICATION OF THALASSEMIA AND HEMOGLOBIN E
Keywords:
Thalassemia, hemoglobin E, infrared spectroscopy, β-thalassemia, α-thalassemiaAbstract
Thalassemia and hemoglobinopathy are forms of hereditary anemia. They are caused by hemoglobin (Hb) disorders. There are many comprehensive and accurate diagnostic methods but they are complex, expensive, and labor-intensive and, in many countries, there are limited facilities to diagnose these diseases. Therefore, the application of the infrared (IR) spectroscopy technique for classification of thalassemias and hemoglobinopathy, especially hemoglobin E (HbE) which is common in Thailand, was studied because it is easy to use and has many advantages: it is reagent free, it uses a small amount of sample (2 μl), and it is simple as it does not require more training. In this study, a total of 124 samples was obtained; 24 samples were normal hemoglobin and there were 4 groups with 20 samples of β+-thalassemia, β0-thalassemia, HbE, α+-thalassemia, and α0-thalassemia hemoglobin, respectively. The results of this study by IR spectroscopy (652 spectra) were analyzed between the normal hemoglobin and the hemoglobin of β+-thalassemia, β0-thalassemia, HbE, α+-thalassemia, and α0-thalassemia using principal component analysis and unsupervised hierarchical cluster analysis of the Unscrambler software. The results found that this technique could be used to classify between the normal hemoglobin and abnormal hemoglobin of all the thalassemias. Among these thalassemic groups, α0-thalassemia was the most differentiated from the normal group, followed by HbE, α+-thalassemia, β+-thalassemia, and β0-thalassemia, respectively.
References
Bain, B.J. (2011). Haemoglobinopathy diagnosis: Algorithms, lessons and pitfalls. Blood Rev., 25(5):205-213.
Bain, B.J., Wild, B.J., Stephens, A.D., and Phelan, L.A. (2010). Globin genes and haemoglobin. In: Variant Haemoglobins: A Guide to Identification. Bain, B.J., Wild, B.J., Stephens, A.D., and Phelan, L.A. (eds). Wiley, NY, USA, p. 1-8.
Bank, A. (1978). The thalassemia syndromes. Blood, 51(3):369-384.
Barrett, A.N., Saminathan, R., and Choolani, M. (2017). Thalassaemia screening and confirmation of carriers in parents. Best Pract. Res. Cl. Ob., 39:27-40.
Barth, A. (2007). Infrared spectroscopy of proteins. Biochim. Biophys. Acta, 1767(9):1073-1101.
Clarke, G.M. and Higgins, T.N. (2000). Laboratory investigation of hemoglobino-pathies and thalassemias: Review and update. Clin. Chem., 46(8):1284-1290.
National Institute of Health of Thailand, (2010). Hemoglobin typing and quantitation. Guideline of laboratory for diagnosis thalassemia and hemoglobinopathy (in Thai). 3rd ed. Nonthaburi: Knockoutblow Press. 27p.
Fucharoen, G., Sanchaisuriya, K., Sae-ung, N., Dangwibul, S., and Fucharoen, S. (2004). A simplified screening strategy for thalassaemia and haemoglobin E in rural communities in south-east Asia. B. World Health Organ., 82:364-372.
Fucharoen, S. and Winichagoon, P. (2010). Prevention and control of thalassemia in Asia. Asian Biomed., 1(1):1-6.
Hartwell, S.K., Srisawang, B., Kongtawelert, P., Christian, G.D., and Grudpan, K. (2005). Review on screening and analysis techniques for hemoglobin variants and thalassemia. Talanta, 65(5):1149-1161.
Huisman, T.H.J. (1992). The β-and δ-thalassemia repository. Hemoglobin, 17(5):479-499.
Liu, K.Z., Tsang, K.S., Li, C.K., Shaw, R.A., and Mantsch, H.H. (2003). Infrared spectroscopic identification of β-thalassemia. Clin. Chem., 49(7):1125-1132.
Ministry of Public Health, Bureau of Policy and Strategy. (2007). Number of in-patients by 75 cause groups according from health service units, whole kingdom: 2003-2012 (in Thai). Available from: http://service. nso.go.th/nso/web/statseries/statseries09.html. Accessed date: June 20, 2014.
Modell, B. and Darlison, M. (2008). Global epidemiology of haemoglobin disorders and derived service indicators. B. World Health Organ., 86:480-487.
Nienhuis, A.W. (1986). Hemoglobin: Molecular, genetic and clinical aspects. Cell, 48(5):731.
Rahmelow, K., Hübner, W., and Ackermann, T. (1998). Infrared absorbances of protein side chains. Anal. Biochem., 257(1):1-11.
Ratanasiri, T. (2007). Prevention and control of thalassemia in obstetric practice. Srinagarind Medical J., 22(4):471- 476. (in Thai)
Rauf, S.E., Shamshad, G.U., Mushtaq, F., Khan, S.A., and Ali, N. (2017). Diagnosing Beta Thalassemia trait in a developing country. Acta Haematologica Polonica, 48(1):18-22.
Scott, A.W., Lutty, G.A., and Goldberg, M.F. (2013). Hemoglobinopathies. In: Retina. Ryan, S.J., Schachat, A.P., Wilkinson, C.P., Hinton, D.R., Sadda, S.R., and Wiedemann, P., (eds). W.B. Saunders & Co., London, UK, p. 1071-1082.
Thumanu, K., Sangrajrang, S., Khuhaprema, T., Kalalak, A., Tanthanuch, W., Pongpiachan, S., and Heraud, P. (2014). Diagnosis of liver cancer from blood sera using FTIR microspectroscopy: A preliminary study. J. Biophotonics, 7(3-4):222-231.
Wan, J.H., Tian, P.L., Yin, H., Han, Y., Wei, X.C., and Pan, T. (2013). A preliminary evaluation of attenuated total reflection Fourier transform infrared spectroscopy for the hematological analysis of thalassemias. Clin. Biochem., 46(1-2):128-132.
Weed, R.I., Reed, C.F., and Berg, G. (1963). Is hemoglobin an essential structural component of human erythrocyte membranes? J. Clin. Invest., 42(1):581-588.
Wibulpolprasert, S. (2011). Public health statistics of number and rate of in-patients (NHSO and CS) according to 298 causes of diseases by all diagnosis, sex and region per 100,000 population. Bureau of Policy and Strategy, Ministry of Public Health, Nonthaburi, Thailand.
Wild, B. and Bain, B.J. (2006). Investigation of abnormal haemoglobins and thalassaemia. In: Dacie and Lewis Practical Haematology. Lewis, S.M., Bain, B.J., Bates, I., and Dacie, J.V., (eds). Churchill Livingstone, Edinburgh, UK, p. 271-310.
