Rapid and reliable β-globin gene cluster haplotyping of sickle cell disease patients by FRET Light Cycler and HRM assays

Joly, Philippe; Lacan, Philippe; Garcia, Cyrielle; Delasaux, Angelique; Francina, Alain

doi:10.1016/j.cca.2011.03.025

Cited by 23 publications

(15 citation statements)

References 11 publications

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“…Conventional methods for large-scale detection of Hb S gene mutations have disadvantages in terms of simplicity, performance, sensitivity, turnaround time and cost. For instance, RFLP assays are time-consuming and are able to lead to misdiagnosis in the case of a supplementary SNP on the restriction sequence (13). Moreover, RFLP and ARMS assays are required for confirmation testing (14).…”

Section: Discussionmentioning

confidence: 99%

Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis

Yue

Lin

Chen

et al. 2014

Molecular Medicine Reports

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Abstract. Each year, ~300,000 individuals with sickle cell disease (SCD), a hemoglobinopathy caused by β-globin gene mutation, are born, and >75% of those are in Africa. The present study examined 511 individuals on the island of Bioko (Equatorial Guinea) and attempted to establish a method for rapid sickle cell disease screening. Following DNA extraction and polymerase chain reaction (PCR) amplification, high resolution melting (HRM) analysis was used to assess the specificity of fluorescence signals of the PCR products and to differentiate various genotypes of these products. The analytical results of HRM were validated using DNA sequencing. By HRM analysis, 80 out of 511 samples were classified as hemoglobin S (Hb S) heterozygotes, while 431 out of 511 samples were classified as wild-type. No mutant homozygote was identified. DNA sequencing indicated that within the 431 wild-type samples as indicated by HRM analysis, one case was actually a Hb S heterozygote and another case was a rare hemoglobin S-C genotype (sickle-hemoglobin C disease). One out of 80 suspected Hb S heterozygotes as indicated by HRM was confirmed as wild-type by DNA sequencing and the results of residual 508 cases were consistent for HRM analysis and sequencing. In conclusion, HRM analysis is a simple, high-efficiency approach for Hb S screening and is useful for early diagnosis of SCD and particularly suitable for application in the African area.

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Section: Discussionmentioning

confidence: 99%

Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis

Yue

Lin

Chen

et al. 2014

Molecular Medicine Reports

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“…UGT1A1 polymorphisms were analyzed by a dedicated high resolution melting (HRM) method (Thomas et al, 2013). The five main SCD haplotypes were identified through genotyping the dedicated HincIIe, XmnI, HindIII G c, HindIII A c, HincIId, HincIIwb, and AvaII SNPs using specific HRM and FRET Light Cycler methods (Joly et al, 2011).…”

Section: Methodsmentioning

confidence: 99%

Genetic modulators of sickle cell disease in French Guiana: Markers of the slave trade

Simonnet

Elanga

Joly

et al. 2016

American J Hum Biol

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“…Deletional α‐thalassamias and β‐globin haplotypes were determined as previously described . Enzyme activity and/or high‐resolution melting (HRM) analysis focusing on the Med and A(−) variants were used to screen for G6PD deficiency.…”

Section: Methodsmentioning

confidence: 99%

G6PD deficiency and absence of α‐thalassemia increase the risk for cerebral vasculopathy in children with sickle cell anemia

Joly

Garnier

Kébaïli

et al. 2015

European J of Haematology

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The aim of this study was to test the association between hematological/genetic factors and cerebral vasculopathy in children with sickle cell anemia (SCA). A group with cerebral vasculopathy (VASC) was composed of children who had stroke (n = 6), silent infarct (n = 11), or an abnormal transcranial Doppler (n = 5). Eighty-four patients had neither positive history of stroke or silent infarct, nor abnormal transcranial Doppler (NORM group). An intermediate group (COND; n = 15) was composed of SCA children with a conditional transcranial Doppler. Biological analyses were performed on samples obtained at steady state and before the beginning of any chronic treatment. The comparisons of the three groups demonstrated a protective effect of α-thalassemia against cerebral vasculopathy through its effects on hemoglobin and reticulocyte levels. Moreover, we observed higher frequency of G6PD deficiency in the VASC group compared with the other groups. Our study confirms the key role of α-thalassemia and G6PD status in the pathophysiology of cerebral vasculopathy in SCA children.

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Rapid and reliable β-globin gene cluster haplotyping of sickle cell disease patients by FRET Light Cycler and HRM assays

Cited by 23 publications

References 11 publications

Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis

Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis

Genetic modulators of sickle cell disease in French Guiana: Markers of the slave trade

G6PD deficiency and absence of α‐thalassemia increase the risk for cerebral vasculopathy in children with sickle cell anemia

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