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Clin Chim Acta. 2013 Sep 23;424:258-60. doi: 10.1016/j.cca.2013.06.023. Epub 2013 Jul 1.

UGT1A1 (TA)n genotyping in sickle-cell disease: high resolution melting (HRM) curve analysis or direct sequencing, what is the best way?

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  • 1Laboratoire de Biochimie et Biologie moléculaire, Unité de pathologie moléculaire du globule rouge, Hôpital Edouard Herriot, Hospices Civils & Université Claude Bernard-Lyon 1, Lyon, France.



Minucci et al. have proposed in 2010 a rapid, simple and cost-effective HRM method on the LightCycler 480® apparatus (Roche) for the determination of the 6/6, 6/7 and 7/7 genotypes of the (TA)n UGT1A1 promoter polymorphism. However, they have not studied the n=5 and n=8 alleles which can be quite frequent in sickle-cell disease patients.


The aim of our study was to test this HRM protocol to all the 10 possible (TA)n UGT1A1 genotypes (i.e. 5/5, 5/6, 5/7, 5/8, 6/6, 6/7, 6/8, 7/7, 7/8 and 8/8) by using our SCD cohort of patients.


All genotypes could be unambiguously identified except 6/7 and 6/8 which give a similar HRM profile. For those two genotypes, the differentiation necessitates either a direct Sanger sequencing or a second PCR protocol followed by a 3% agarose gel migration.


For the (TA)n UGT1A1 promoter genotyping of African patients, each lab has to wonder what is the best way between (i) direct Sanger sequencing of all patients and (ii) HRM protocol for all patients followed by a complementary analysis to differentiate the 6/7 and 6/8 genotypes.

© 2013. Published by Elsevier B.V. All rights reserved.


Gilbert's syndrome; HRM; Sanger sequencing; Sickle-cell disease; UGT1A1 promoter

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