DNA based-tests; buccal swab; sickle cell anemia; umbilical cord blood; venous blood; Immunology and Allergy; Hematology; Public Health, Environmental and Occupational Health; Clinical Biochemistry; Medical Laboratory Technology; Biochemistry (medical); Microbiology (medical)
Abstract :
[en] BACKGROUND: Hemoglobin-based tests form the reference diagnostic test for SCA. In limited resource countries, these tests face limitations including cost, low sensitivity due to recurrent transfusions in endemic malaria region, and interference from fetal hemoglobin in neonatal diagnostic. This study aimed at adapting DNA-based SCA tests to limited resource countries and evaluating the economic benefit.
METHODS: 338 participants were recruited in the Democratic Republic of Congo, sorted in 3 cohorts based on venous blood, umbilical cord blood (UCB) and buccal swab sampling. RFLP was performed to identify mutated allele. The feasibility and technical validity of this RFLP was evaluated for specimens collected on DBS cards and on EDTA tubes. RFLP on DBS stored at room temperature was regularly repeated to assess sample conservation. Finally, the cost analysis was performed.
RESULTS: DBS cards yielded identical results to extracted DNA. Repeated testing returned the same result after four years. The DBS-based test performed on UCB or on buccal swab had a sensitivity and a precision of 100%. Cost comparison indicated that our approach costs half price of the widely used isoelectrofocussing of hemoglobin.
CONCLUSION: The implemented DNA-based test approach overcomes the limitations faced by hemoglobin-based tests, while being more affordable. We propose to implement the RFLP test as a first line diagnostic test after transfusion and as second tiers for newborn screening. However, users should be aware that this test is unable to differentiate HbC from HbS or identify other point mutation of gene deletion of HBB gene.
Research Center/Unit :
GIGA-AUHG - GIGA Cancer-Human Genetics - ULiège
Disciplines :
Laboratory medicine & medical technology
Author, co-author :
Ngole, Mamy ; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium ; Center for Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Department of Medical Biology, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Race, Valerie; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium
Mbayabo, Gloire ; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium ; Center for Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Department of Pediatrics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Lumbala, Paul; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium ; Center for Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Department of Pediatrics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Songo, Cathy; Center for Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Lukusa, Prosper Tshilobo; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium ; Center for Human Genetics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo ; Department of Pediatrics, Faculty of Medicine, University of Kinshasa, Kinshasa, Democratic Republic of Congo
Devriendt, Koenraad; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium
Matthijs, Gert; Center for Human Genetics, Faculty of Medicine, KU Leuven, Leuven, Belgium
Lumaka Zola, Aimé ; Centre Hospitalier Universitaire de Liège - CHU ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Génétique humaine ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > GIGA-R : Génétique humaine
Language :
English
Title :
DNA testing for sickle cell anemia in Africa: Implementation choices for the Democratic Republic of Congo.
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.
Grosse SD, Odame I, Atrash HK, Amendah DD, Piel FB, Williams TN. Sickle cell disease in Africa: a neglected cause of early childhood mortality. Am J Prev Med. 2011;41(6 Suppl 4):S398-S405.
Kitenge R, Tshilolo L, Loko G, Wamba G, Gonzalez JP. Diagnostic tools and follow-up of sickle-cell anemia in Central Africa. Med Sante Trop. 2018;28(2):124-127.
Piccin A, Fleming P, Eakins E, McGovern E, Smith OP, McMahon C. Sickle cell disease and dental treatment. J Ir Dent Assoc. 2008;54(2):75-79.
Vandepitte J, Stijns J. Hemoglobinoses in the Congo (Léopoldville) and in Ruanda-Urundi. Ann Soc Belg Med Trop. 1963;43:271-281.
Bardakdjian-Michau J, Dhondt L, Ducrocq R, et al. Good practices for the study of hemoglobin. Ann Biol Clin. 2003;61(4):401-409.
Tshilolo L, Mukendi R, Girot R. Sickle cell anemia in the south of Zaire. Study of two series of 251 and 340 patients followed-up 1988–1992. Arch Pediatr. 1996;3(2):104-111.
Shongo MY, Mukuku O, Lubala TK, et al. Sickle cell disease in stationary phase in 6–59 months children in Lubumbashi: epidemiology and clinical feature. Pan Afr Med J. 2014;19:71.
El-Haj N, Hoppe C. Newborn Screening for SCD in the USA and Canada. Int J Neonatal Screen. 2018;4:36.
Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988;16(3):1215.
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):e22140.
Marshall CR, Chowdhury S, Taft RJ, et al. Best practices for the analytical validation of clinical whole-genome sequencing intended for the diagnosis of germline disease. NPJ Genom Med. 2020;5:47.
Leeflang MM, Rutjes AW, Reitsma JB, Hooft L, Bossuyt PM. Variation of a test's sensitivity and specificity with disease prevalence. CMAJ. 2013;185(11):E537-E544.
Chacon-Cortes D, Haupt LM, Lea RA, Griffiths LR. Comparison of genomic DNA extraction techniques from whole blood samples: a time, cost and quality evaluation study. Mol Biol Rep. 2012;39(5):5961-5966.
Mas S, Crescenti A, Gasso P, Vidal-Taboada JM, Lafuente A. DNA cards: determinants of DNA yield and quality in collecting genetic samples for pharmacogenetic studies. Basic Clin Pharmacol Toxicol. 2007;101(2):132-137.
Fata A, Khamesipour A, Mohajery M, et al. Whatman paper (FTA Cards) for storing and transferring Leishmania DNA for PCR examination. Iran J Parasitol. 2009;4(4):1.
Abdelwhab EM, Luschow D, Harder TC, Hafez HM. The use of FTA(R) filter papers for diagnosis of avian influenza virus. J Virol Methods. 2011;174(1–2):120-122.
Rabodoarivelo MS, Imperiale B, Andrianiavomikotroka R, et al. Performance of four transport and storage systems for molecular detection of multidrug-resistant tuberculosis. PLoS One. 2015;10(10):e0139382.
Dong L, Lin C, Li L, et al. An evaluation of clinical performance of FTA cards for HPV 16/18 detection using cobas 4800 HPV Test compared to dry swab and liquid medium. J Clin Virol. 2017;94:67-71.
Siegel CS, Stevenson FO, Zimmer EA. Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa. Appl. Plant Sci. 2017;5(2):1600109.
Burton GJ, Sebire NJ, Myatt L, et al. Optimising sample collection for placental research.". Placenta. 2014;35(1):9-22.
Abraham JE, Maranian MJ, Spiteri I, et al. Saliva samples are a viable alternative to blood samples as a source of DNA for high throughput genotyping. BMC Med Genomics. 2012;5:19.
Rajendram D, Ayenza R, Holder FM, Moran B, Long T, Shah HN. Long-term storage and safe retrieval of DNA from microorganisms for molecular analysis using FTA matrix cards. J Microbiol Methods. 2006;67(3):582-592.
Peluso AL, Cascone AM, Lucchese L, et al. Use of FTA cards for the storage of breast carcinoma nucleic acid on fine-needle aspiration samples. Cancer Cytopathol. 2015;123(10):582-592.
Barth H, Morel A, Mougin C, et al. Long-term storage and safe retrieval of human papillomavirus DNA using FTA elute cards. J Virol Methods. 2016;229:60-65.
Corradini B, Alu M, Magnanini E, Galinier ME, Silingardi E. The importance of forensic storage support: DNA quality from 11-year-old saliva on FTA cards. Int J Legal Med. 2019;133(6):1743-1750.
Piccin A. Do we need to test blood donors for sickle cell anaemia? Blood Transfus. 2010;8(3):137-138.
