BACKGROUND: β-Globin haplotypes are important to predict the clinical development of patients suffering from sickle cell disease (SCD). Five main haplotypes (Benin, Bantu, Senegal, Cameroon and Arabic-Indian) are defined for β(S) chromosomes and their determination usually requires the genotyping by restriction fragment length polymorphism (RFLP) of six to eight single nucleotide polymorphisms (SNPs). However, RFLP is time-consuming and can lead to a misdiagnosis in case of a supplementary SNP on the restriction sequence. We propose a rapid β-globin haplotyping method using fluorescence resonance transfer (FRET) and high resolution melting (HRM) assays. METHODS: We have settled a fluorescence resonance energy transfer (FRET) assay for HincII ε, XmnI, HindIII (G)γ, HindIII (A)γ, HincII δ and a high resolution melting (HRM) assay for HincII ψβ. These six SNPs are sufficient in most cases to determine the β(S) haplotype. RESULTS: Our methodology allowed us to successfully determine the β-globin haplotypes of 139 patients suffering from sickle cell disease. For some β(S) / β(0)-patients, a supplementary SNP has been identified on the HindIII (G)γ restriction sequence leading to a false-negative RFLP result. CONCLUSION: Combination of FRET and HRM assays is a rapid and reliable method for the β-globin gene cluster haplotyping.
BACKGROUND: β-Globin haplotypes are important to predict the clinical development of patients suffering from sickle cell disease (SCD). Five main haplotypes (Benin, Bantu, Senegal, Cameroon and Arabic-Indian) are defined for β(S) chromosomes and their determination usually requires the genotyping by restriction fragment length polymorphism (RFLP) of six to eight single nucleotide polymorphisms (SNPs). However, RFLP is time-consuming and can lead to a misdiagnosis in case of a supplementary SNP on the restriction sequence. We propose a rapid β-globin haplotyping method using fluorescence resonance transfer (FRET) and high resolution melting (HRM) assays. METHODS: We have settled a fluorescence resonance energy transfer (FRET) assay for HincII ε, XmnI, HindIII (G)γ, HindIII (A)γ, HincII δ and a high resolution melting (HRM) assay for HincII ψβ. These six SNPs are sufficient in most cases to determine the β(S) haplotype. RESULTS: Our methodology allowed us to successfully determine the β-globin haplotypes of 139 patients suffering from sickle cell disease. For some β(S) / β(0)-patients, a supplementary SNP has been identified on the HindIII (G)γ restriction sequence leading to a false-negative RFLP result. CONCLUSION: Combination of FRET and HRM assays is a rapid and reliable method for the β-globin gene cluster haplotyping.
Authors: Tosséa A Stéphane Koui; Alloh Albert Gnondjui; Adji Eric Gbessi; Ako Aristide Bérenger Ako; Baba Coulibaly; A Delpêche Aka; Bi Sery E Gonedele; Offiana André Toure; Ronan Jambou Journal: BMC Med Genomics Date: 2022-05-23 Impact factor: 3.622
Authors: Seosamh Ó Lochlainn; Stephen Amoah; Neil S Graham; Khalid Alamer; Juan J Rios; Smita Kurup; Andrew Stoute; John P Hammond; Lars Østergaard; Graham J King; Phillip J White; Martin R Broadley Journal: Plant Methods Date: 2011-12-08 Impact factor: 4.993
Authors: Elmutaz M Shaikho; John J Farrell; Abdulrahman Alsultan; Hatem Qutub; Amein K Al-Ali; Maria Stella Figueiredo; David H K Chui; Lindsay A Farrer; George J Murphy; Gustavo Mostoslavsky; Paola Sebastiani; Martin H Steinberg Journal: BMC Genomics Date: 2017-08-11 Impact factor: 3.969