Genotyping of S and C Mutated Beta Globin Gene: Development of a Set of Primers for Use with DfV'VnG PCR Systems

International audienceSickle cell disease is a genetic disorder that affects nearly 5% of world population. In ivory coast, SCD is a real problem of health and screening is not systematic after born. Here, we designed a set of primers to detect abnormal hemoglobin S and C which can be used both in conventional and quantitative PCR (by curves combinations analysed). A total of 60 blood samples including 13 AA, 23 AS, 9 SS, 12 SC and 3 CC hemoglobin type were screened using hemoglobin electrophoresis and PCR. The universal control primer HBU/R4 was successfully amplified for all of 60 samples. In conventional PCR, for typing of allele S sensibility and specificity of primers were respectively 86.36% and 87.50%. For allele C, sensibility and specificity of this pair were respectively 53.33% and 91.11%. In qPCR, specificity and sensitivity of primers were greater than 85% for allele S and C specific primers

Artemisinin derivative-containing therapies and abnormal hemoglobin: Do we need to adapt the treatment?

International audienceBackground: Artemisinin-based treatment in malaria patients with abnormal hemoglobin may be ineffective because of their genetic particularity, which could lead to resistance. The main purpose of this study was to assess the effect of artemisinin derivatives on in vivo parasite clearance according to erythrocyte variants. In vivo response was investigated through retrospective data obtained over a 42-day artemether-lumefantrine/artesunate amodiaquine efficacy protocol conducted from 2012 to 2016. Results: A total of 770 patients in Côte d’Ivoire attending the hospitals of Anonkoua-koute (Abidjan), Petit Paris (Korhogo), Libreville (Man), Dar es salam (Bouaké), Ayamé and Yamoussoukro with acute uncomplicated falciparum malaria were selected for successful hemoglobin typing. HbAS, HbSS, HbAC, and HbSC genotypes were found. Parasite clearance time was obtained for 414 patients. In the population with abnormal hemoglobin, parasite densities on admission and parasite clearance rates were significantly lower in the HbSC group compared to HbAA (p = 0.02 and p = 0.007, respectively). After PCR correction on day 42, the acute treatment rate was 100% for each group. Parasite half-life and time for initial parasitaemia to decline by 50 and 99% were longer for the HbSC group (p < 0.05). The study also investigated the prevalence of K13-propeller polymorphisms across different hemoglobin genotype groups. A total of 185 and 63 samples were sequenced in the HbAA group and patients with abnormal Hb, respectively. Only two nonsynonymous mutations D559N and V510M were found in the HbAA group. Conclusion: Although this study proved good efficacy of artemether-lumefantrine and artesunate amodiaquine in the treatment of uncomplicated Plasmodium falciparum malaria in patients with abnormal hemoglobin, the increased delay of parasite clearance may represent a threat to health in these patients in relation with sickle cell crisis, which could support selection of parasites resistant to artemisinin

High level of heterozygous haplotype of hemoglobin in Abidjan population with mild malaria

International audienceBackgroundSickle cell disease (SCD) is a hemoglobin disorders that concern 300,000 newborns each year around the world. There are hemoglobin haplotypes that affect SCD clinic expression.MethodsOur goal was to identify the hemoglobin’s haplotypes among individuals with mild malaria independently of SCD status in Côte d’Ivoire. To determine these haplotypes, specific restriction enzyme (RE) is used after PCR amplification with each primer. According to the digestion of PCR product by RE, five hemoglobin’s haplotypes are found in the world.ResultsIn Côte d’Ivoire, no study has yet deeply described the distribution of haplotypes. Four different “classical” haplotypes of hemoglobin were detected: Benin (56.5%), Bantou (28.5%), Senegal (4%), Cameroun (1%); and 10% of atypical profiles. Heterozygous haplotype (69%) were more frequent than homozygous haplotype (31%).ConclusionsIn this preliminary study, we note a high prevalence of atypical and heterozygous haplotype. Benin haplotype that is associated with severity of SCD was most predominant in our studied population