Design and Construction of a Low-Cost Hemoglobin Electrophoresis (Genotype) Machine for the Diagnosis of Inherited Genotype Disorder

Oluwasegun Temitope Adefemi, Vincent Andrew Akpan, Jessica Temilola Ovabor, Olabisi Glory Adekoya, Buniechukwuenu Emmanuel Mbamalu

Abstract

Designing and constructing a low-cost haemoglobin electrophoresis (genotype) machine using cellulose acetate paper is an exciting project that underscores using locally sourced materials and resources to achieve its aims. The technique of haemoglobin electrophoresis is adequately harnessed to detect and evaluate various types of haemoglobin disorder, which include sickle anaemia and many other abnormal genotypes. The cost of acquisition and maintenance of the equipment is made available at a comparatively reduced rate by using materials and components that are remotely accessed to construct the haemoglobin electrophoresis machine, which is efficient and cost-effective. The design and construction team incorporated a power supply unit, buffer solution (tris), cellulose acetate paper, filter paper, electrodes (anode and cathode), switch, cable, capacitor, fuse, and other necessary components. The successfully constructed genotype machine was validated for effective and efficient diagnostic purposes for detecting different types of haemoglobin genotypes by testing it using several specimens of known haemoglobin genotypes of different types. The researchers compared the results from this testing process with those obtained from a standardised haemoglobin genotype machine. The test result obtained from this verification and validation process showcases that the low-cost haemoglobin electrophoresis ma-chine using cellulose acetate paper can effectively separate, distinguish and identify different haemoglobin variants such as HBAA, HBAS, HBAC, HBCC, HBSS, HBSC, HBF, HBAC, HBAF with few limitations. The drawbacks include the inability to distinguish other haemoglobin genotype variants that co-migrate with HBAA, HBAS, HBAC, HBCC, HBSS, HBSC, HBF, HBAC, and HBAF when exposed to the electric field in the electrophoretic chamber. Examples of such haemoglobin genotypes include HB E, HB D, HB O, and HB G, with many other haemoglobin variants having exact migration patterns on the cellulose acetate paper when subjected to the electric field. However, this limitation can be overlooked as these haemoglobin variants are not peculiar to people in this part of the world (Nigeria) where the project has been carried out. Hence, the design and construction of this haemoglobin electrophoresis machine will give people from low-resource areas easy access to carry out the test and enhance prompt detection and management of hemoglobinopathies.




Keywords


Hemoglobin; Electrophoresis; Cellulose Acetate Paper; Hemoglobin Electrophores; Hemoglobinopathy; Validation; Hemoglobin-electrophoresis; Genotype.

Full Text:

PDF


References


1. Arishi, W. A., Alhadrami, H. A., & Zourob, M. (2021). Techniques for the Detection of Sickle Cell Disease: A review. Micromachines, 12(5), 519. doi: 10.3390/mi12050519

2. World Health Organization. (2012). Recommendations for management of common childhood conditions. Retrieved from https://iris.who.int/bitstream/handle/10665/44774/9789241502825_eng.pdf?sequence

3. Harteveld, C. L., Achour, A., Arkesteijn, S. J. G., Ter Huurne, J., Verschuren, M., Bhagwandien‐Bisoen, S., Schaap, R., Vijfhuizen, L., Idrissi, H. E., & Koopmann, T. T. (2022). The hemoglobinopathies, molecular disease mechanisms and diagnostics. International Journal of Laboratory Hematology, 44(S1), 28–36. doi: 10.1111/ijlh.13885

4. Tahura, S. (2017). Thalassemia and other Hemoglobinopathies in Bangladeshi Children. Imperial Journal of Interdisciplinary Research (IJIR), 3(4)

5. Kato, G. J., Piel, F. B., Reid, C. D., Gaston, M. H., Ohene-Frempong, K., Krishnamurti, L., Smith, W. R., Panepinto, J. A., Weatherall, D. J., Costa, F. F., & Vichinsky, E. P. (2018). Sickle cell disease. Nature Reviews Disease Primers, 4(1). doi: 10.1038/nrdp.2018.10

6. Sridhar, А. & Idowu, M. (2021). Hemoglobin Electrophoresis in Sickle Cell Disease: A Primer for the Clinician. Retrieved from https://www.hematology.org/education/trainees/fellows/hematopoiesis/2021/hemoglobin-electrophoresis-in-sickle-cell-disease

7. Khosa, S. M., Usman, M., Moinuddin, M., Mehmood, H. O., & Qamar, K. (2015b). Comparative analysis of cellulose acetate haemoglobin electrophoresis and high-performance liquid chromatography for quantitative determination of haemoglobin A2. Blood Research, 50(1), 46. doi: 10.5045/br.2015.50.1.4

8. Keren, D. (2003). Protein electrophoresis in clinical diagnosis. In CRC Press eBooks. doi: 10.1201/b13302

9. Mosca, G. W. G. D. A., Branca, M. T., Carta, M., Genna, M. L., Giorda, C. B., Ghidelli, R., Ghislandi, G., Iafusco, D., Lapolla, A., Lombardi, V. B., Scher, C. a. L., Marra, M., Medea, G., Meschi, F., Pizzini, A., Rossi, F., Scalpone, R., Tofini, G., Trovati, M., & Zaninotto, M. (2010). Recommendations for the implementation of international standardisation of glycated haemoglobin in Italy. Clinical Chemistry and Laboratory Medicine (CCLM), 48(5), 623–626. doi: 10.1515/cclm.2010.1409

10. Schinzel, A. (2015). Genetics and Genomics in Medicine. European Journal of Human Genetics, 23(5), 719. doi: 10.1038/ejhg.2015.18

11. Nagel, R. L., Fabry, M. E., Pagnier, J., Zohoun, I., Wajcman, H., Baudin, V., & Labie, D. (1985). Hematologically and genetically distinct forms of sickle cell anaemia in Africa. New England Journal of Medicine, 312(14), 880–884. doi: 10.1056/nejm198504043121403

12. Association of Public Health Laboratories. (2015). Hemoglobinopathies: Current Practices For Screening, Confirmation And Follow-Up. Retrieved from https://www.cdc.gov/sickle-cell/media/pdfs/nbs_hemoglobinopathy-testing_122015.pdf

13. Bain, B. J. (2011). Haemoglobinopathy diagnosis: Algorithms, lessons and pitfalls. Blood Reviews, 25(5), 205–213. doi: 10.1016/j.blre.2011.04.001

14. Ghosh, K., Colah, R., Choudhry, V., Das, R., Manglani, M., Madan, N., Saxena, R., Jain, D., Marwaha, N., Mohanty, D., Ghosh, M., Ross, C., Verma, I., Agarwal, S., & Choudhary, R. (2014). Guidelines for screening, diagnosis and management of hemoglobinopathies. Indian Journal of Human Genetics, 20(2), 101. doi: 10.4103/0971-6866.142841

15. Pum, J. (2019). A practical guide to validation and verification of analytical methods in the clinical laboratory. Advances in Clinical Chemistry, 215–281. doi: 10.1016/bs.acc.2019.01.006

16. Breadmore, M. C., Grochocki, W., Kalsoom, U., Alves, M. N., Phung, S. C., Rokh, M. T., Cabot, J. M., Ghiasvand, A., Li, F., Shallan, A. I., Keyon, A. S. A., Alhusban, A. A., See, H. H., Wuethrich, A., Dawod, M., & Quirino, J. P. (2018). Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2016–2018). Electrophoresis, 40(1), 17–39. doi: 10.1002/elps.201800384

17. El-Beshlawy, A., & Youssry, I. (2009). Prevention of hemoglobinopathies in Egypt. Hemoglobin, 33(sup1), S14–S20. doi: 10.3109/03630260903346395

18. Kunz, J. B., & Kulozik, A. E. (2020). Gene therapy of the hemoglobinopathies. HemaSphere, 4(5). doi: 10.1097/hs9.0000000000000479

19. Kumar, R., & Derbigny, W. A. (2019). Cellulose acetate electrophoresis of haemoglobin. In B. Kurien, & R. Scofield, Electrophoretic separation of proteins: methods and protocols. New York: Humana Press.

20. Rentapalli, B. R., Gade, J., Sulemani, D., MD, Ganji, S. B., Thandu, P. K., Revupalli, S., Kavali, S., & Ramtenki, A. (2017). Agar gel alkaline electrophoresis as a cost-effective screening modality for abnormal haemoglobins. IOSR Journal of Dental and Medical Sciences, 16(04), 39–43. doi: 10.9790/0853-1604083943

21. Salih, K. M. A., Saad, A. A., Eldouch, W. M., Mohamed, H. K., Alfaifi, J., Andarawi, M. O., & Elshayeb, A. A. (2022). Evaluation of current clinical research spectrum of sickle cell anaemia to approach new innovative developmental techniques for enhancing Sudanese health systems. Research Square (Research Square). doi: 10.21203/rs.3.rs-2107299/v1

22. Khosa, S. M., Usman, M., Moinuddin, M., Mehmood, H. O., & Qamar, K. (2015). Comparative analysis of cellulose acetate haemoglobin electrophoresis and high-performance liquid chromatography for quantitative determination of haemoglobin A2. Blood Research, 50(1), 46. doi: 10.5045/br.2015.50.1.46

23. Greene, D. N., Vaughn, C. P., Crews, B. O., & Agarwal, A. M. (2015). Advances in detection of hemoglobinopathies. Clinica Chimica Acta, 439, 50–57. doi: 10.1016/j.cca.2014.10.006

24. Ilyas, S., Simonson, A. E., & Asghar, W. (2020). Emerging point-of-care technologies for sickle cell disease diagnostics. Clinica Chimica Acta, 501, 85–91. doi: 10.1016/j.cca.2019.10.025

25. Alapan, Y., Fraiwan, A., Kucukal, E., Hasan, M. N., Ung, R., Kim, M., Odame, I., Little, J. A., & Gurkan, U. A. (2016). Emerging point-of-care technologies for sickle cell disease screening and monitoring. Expert Review of Medical Devices, 13(12), 1073–1093. doi: 10.1080/17434440.2016.1254038

26. Al-Allawi, N., Allawi, S. A., & Jalal, S. D. (2020). Genetic epidemiology of hemoglobinopathies among Iraqi Kurds. Journal of Community Genetics, 12(1), 5–14. doi: 10.1007/s12687-020-00495-z

27. Singh, P. J., Shrivastava, A. C., & Shrikhande, A. V. (2014). Prenatal diagnosis of sickle cell disease by the technique of PCR. Indian Journal of Hematology and Blood Transfusion, 31(2), 233–241. doi: 10.1007/s12288-014-0427-8

28. Ghosh, K., Ghosh, K., Agrawal, R., & Nadkarni, A. H. (2019). Recent advances in screening and diagnosis of hemoglobinopathy. Expert Review of Hematology, 13(1), 13–21. doi: 10.1080/17474086.2019.1656525

29. Ullah, Z., Rasool, R., Aziz, N., Bano, R., Bashir, S., Ali, S. A., & Khattak, A. A. (2022). Spectrum of Inherited Hemoglobin Disorder in Patients Presenting for Hb Electrophoresis: A Single Center Study in District Dera Ismail Khan. Pakistan Journal of Medical & Health Sciences, 16(7), 151–153. doi: 10.53350/pjmhs22167151

30. Genotype Machine. (n. d.). Genotype Machine. Retrieved from https://clinicaid.com.ng/product/genotype-machine/

31. Das, R., & Sharma, P. (2019). Disorders of abnormal haemoglobin. In Elsevier eBooks (pp. 327–339). doi: 10.1016/b978-0-12-809356-6.00018-6

32. Costa, F. F., & Conran, N. (2016). Sickle cell anaemia. In Springer eBooks. doi: 10.1007/978-3-319-06713-1

33. Chou, S. T. (2018). Red blood cell antigen genotyping for haemoglobinopathies: considerations for thalassaemia and sickle cell disease. ISBT Science Series, 13(3), 238–243. doi: 10.1111/voxs.12416

34. Kar, A. (2021). Birth defects in India. In Springer eBooks. doi: 10.1007/978-981-16-1554-2

35. Akinyanju, O. O., Otaigbe, A. I., & Ibidapo, M. O. O. (2005). Outcome of holistic care in Nigerian patients with sickle cell anaemia. Clinical & Laboratory Haematology, 27(3), 195–199. doi: 10.1111/j.1365-2257.2005.00683.x

36. Samavat, A., & Modell, B. (2004). Iranian National Thalassaemia Screening Programme. BMJ, 329(7475), 1134–1137. doi: 10.1136/bmj.329.7475.1134

37. Westermeier, R. (2016). Electrophoresis in practice. doi: 10.1002/9783527695188


Article Metrics

Metrics Loading ...

Metrics powered by PLOS ALM

Refbacks

  • There are currently no refbacks.




Copyright (c) 2024 Oluwasegun Temitope Adefemi, Vincent Andrew Akpan, Jessica Temilola Ovabor, Olabisi Glory Adekoya, Buniechukwuenu Emmanuel Mbamalu

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.