Central Africa; hemoglobin electrophoresis; molecular testing; sickle-cell anemia; Microbiology (medical); Biochemistry (medical); Medical Laboratory Technology; Clinical Biochemistry; Public Health, Environmental and Occupational Health; Hematology; Immunology and Allergy
Abstract :
[en] BACKGROUND: Sickle-cell anemia (SCA) is the most common genetic disease worldwide caused by a single mutation in the gene HBB. DNA testing can help to clarify the diagnosis when Hb electrophoresis is inconclusive. We evaluated the usefulness and feasibility of DNA-based diagnosis of SCA in rural Central Africa.
METHODS: This is a cross-sectional study conducted from November 2016 to end October 2017 in the Hôpital Saint Luc de Kisantu, located 120 km from Kinshasa. This hospital offers the management of SCA patients, mainly identified using the Sickling test (Emmel test) combined with clinical features. We included patients aged 6 months to 18 years locally diagnosed as SCA, and we collected clinical and hematological data. All patients were offered Hb electrophoresis and DNA testing at the Center for Human Genetics of the University of Kinshasa.
RESULTS: This study included 160 patients. Hemoglobin capillary electrophoresis suggested that 136 (85%) were homozygote SS, 13 (8.1%) were heterozygote (AS), and 11 (6.9%) were homozygote normal (AA). DNA testing confirmed these electrophoresis findings, with the exception of four patients, two AS in electrophoresis were found SS due to recent transfusion, and two SS in electrophoresis were found AS because they have compound heterozygous form S/β°-thalassemia. The diagnosis of SCA was therefore wrongly ascertained with Emmel test in 15% of patients.
CONCLUSION: This study reveals a high proportion of false-positive SCA diagnoses in a rural environment in Central Africa. This underlines the importance of DNA testing in conjunction with Hb electrophoresis.
Disciplines :
Genetics & genetic processes
Author, co-author :
Mbayabo, Gloire ; Department of Pediatrics, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium ; Center of Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Lumbala Kabuyi, Paul; Department of Pediatrics, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium ; Center of Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Ngole, Mamy ; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium ; Center of Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Department of Clinical Biology, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Lumaka Zola, Aimé ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Génétique humaine ; Department of Pediatrics, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Center of Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Race, Valerie; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium
Matthijs, Gert; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium
Minga, Tite Mikobi; Center of Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Département des sciences de base, Laboratory of biochemistry and molecular biology, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Devriendt, Koenraad; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium
Van Geet, Chris; Department of Cardiovascular Sciences and Pediatrics (Hemato-oncology), KU Leuven and University Hospitals Leuven, Leuven, Belgium
Tshilobo, Prosper Lukusa; Department of Pediatrics, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Center for Human Genetics, KU Leuven and University Hospitals Leuven, Leuven, Belgium ; Center of Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Language :
English
Title :
Value of DNA testing in the diagnosis of sickle-cell anemia in childhood in an environment with a high prevalence of other causes of anemia.
Livingstone FB. Frequencies of Hemoglobin Variants. Vol 148. Oxford University Press; 1985:148-162.
Shongo MY, Mukuku O, Lubala TK, et al. Sickle cell disease in stationary phase in 6-59 months children in Lubumbashi: epidemiology and clinical features. Pan Afr Med J. 2014;19:71.
Cotton F, Lin C, Fontaine B, Gulbis B, Janssens J, Vertongen F. Evaluation of a capillary electrophoresis method for routine determination of hemoglobins A2 and F. Clin Chem. 1999;45(2):237-243.
Orts JA, Zúñiga Á, Bello Y, Fabregat AB, Vicente AI. Hb A1c determination by capillary electrophoresis is an efficient method for detecting β-Thalassemias and hemoglobin variants. Hemoglobin. 2016;40(5):335-340.
Wong P, Weerakul J, Sritippayawan S. Hemoglobin analysis in the first year of life. Mediterr J Hematol Infect Dis. 2016;8(1):1-6.
Tshilolo L, Kafando E, Sawadogo M, et al. Neonatal screening and clinical care programmes for sickle cell disorders in sub-Saharan Africa: lessons from pilot studies. Public Health. 2008;122(9):933-941.
WHO. Malaria Elimination: a Field Manual for Low and Moderate Endemic Ocuntries. WHO Library Cataloguing-in-Publication Data; 2007.
Ferrari G, Ntuku HM, Schmidlin S, Diboulo E, Tshefu AK, Lengeler C. A malaria risk map of Kinshasa, Democratic Republic of Congo. Malar J. 2016;15(1):1-16.
Sumbele IUN, Kimbi HK, Ndamukong-Nyanga JL, et al. Malarial anaemia and anaemia severity in apparently healthy primary school children in urban and rural settings in the mount cameroon area: cross sectional survey. Paul R, Editor PLoS One [Internet]. 2015;10(4):e0123549. doi:10.1371/journal.pone.0123549
Arishi WA, Al-hadrami HA, Zourob M. Techniques for the detection of sickle cell disease: a review. Micromachines. 2021;12(5):1-22.
Ballas SK, Kesen MR, Goldberg MF, et al. Beyond the definitions of the phenotypic complications of sickle cell disease: an update on management. Sci World J. 2012;2012:1-55.
Mikobi TM, Lukusa Tshilobo P, Aloni MN, Akilimali PZ, Mvumbi-Lelo G, Mbuyi-Muamba JM. Clinical phenotypes and the biological parameters of Congolese patients suffering from sickle cell anemia: a first report from Central Africa. J Clin Lab Anal. 2017;31(6):1-6.
Schmugge M, Waye JS, Basran RK, Zurbriggen K, Frischknecht H. THE Hb S/β + -thalassemia phenotype demonstrates that THE IVS-I (−2) (a>C) mutation is a mild β-thalassemia allele. Hemoglobin. 2008;32(3):303-307. doi:10.1080/03630260802004459
Broquere C, Brudey K, Harteveld CL, et al. Phenotypic expression and origin of the rare β-thalassemia splice site mutation HBB:c.315 + 1G>T. Hemoglobin. 2010;34(3):322-326. doi:10.3109/03630269.2010.484956
Tshilolo L, Aissi LM, Lukusa D, et al. Neonatal screening for sickle cell anaemia in The Democratic Republic of the Congo: experience from a pioneer project on 31 204 newborns. J Clin Pathol. 2009;62(1):35-38.
Agasa B, Bosunga K, Opara A, et al. Prevalence of sickle cell disease in a northeastern region of the Democratic Republic of Congo: what impact on transfusion policy? Transfus Med. 2010;20(1):62-65.
Herrick JB. Peculiar elongated and sickle-shaped red blood corpuscles in a case of severe anemia. Arch Intern Med. 1910;VI(5):517-521.
Emmel VE. A study of the erythrocytes in a case of severe anemia with elongated and sickle-shaped red blood corpuscles. Arch Intern Med. 1917;XX(4):586.
Badawi M, Garoot S, Tahlawi M, Hindawi S, Adam S. Evaluation of a point-of-care method for screening blood donors for sickle cell status. Transfusion. 2020;60(Suppl 1):S10-S14.
Fakher R, Bijan K, Taghi A. Application of diagnostic methods and molecular diagnosis of hemoglobin disorders in Khuzestan province of Iran. Indian J Hum Genet. 2007;13(1):5-15. http://www.ijhg.com/text.asp?2007/13/1/5/32028
Serjeant GR. Sickle Cell Disease. Oxford Univ Press; 1986:478.
Ragusa A, Amata S, Amata S, et al. Asymptomatic and mild beta-thalassemia in homozygotes and compound heterozygotes for the IVS2+1G-->A mutation: role of the beta-globin gene haplotype. 2003;88(10):1099-1105. https://pubmed.ncbi.nlm.nih.gov/14555304/ Accessed 2021 Dec 13.
Dedoussis GVZ, Mandilara GD, Boussiu M, Loutradis A. HbF production in β thalassaemia heterozygotes for the IVS-II-1 G→a β0-globin mutation. Implication of the haplotype and the (G)γ-158 C→T mutation on the HbF level. Am J Hematol. 2000;64(3):151-155.
Kluge ML, Hoyer JD, Swanson KC, Oliveira JL. β-thalassemia major resulting from compound heterozygosity for HBB: c.92+2T>C [formerly known as IVS-I-2 (T>C)] and a novel β(0)-thalassemia frameshift mutation: HBB: c.209delG; p.Gly70Valfs*20. Hemoglobin. 2014;38(4):292-294. https://www.tandfonline.com/action/journalInformation?journalCode=ihem20 Accessed 2021 Dec 4.
Quinn CT, Shull EP, Ahmad N, Rogers ZR, Buchanan GR. Prognostic significance of early Vaso-occlusive complications in children who have sickle cell anemia. Blood. 2005;106(11):3176. 10.1182/blood.V106.11.3176.3176
