2009•Leukemia & lymphoma/Leukemia and lymphomaRequires access
Detection of mutations inGATA1gene using automated denaturing high-performance liquid chromatography and direct sequencing in children with Down syndrome
Márcia Rodrigues Amorim, Márcia Rodrigues Amorim, Alexandre Figueiredo, Alessandra Splendore, Isis Quezado Magalhães, Maria S. Pombo‐de‐Oliveira, Bcsgail, Kênia Balbi El-Jaick, Maria Lydia d'Andréa, Jozina Aquino, Dora Márcia Alencar, Sílvia Regina Brandalise, Lilian Burlemaqui, Teresa Cristina Cardoso, Eni Guimarães Carvalho, Virgínia Maria Cóser, Imaruí Costa, Dolores Dorea, Maurício Drumond, Venâncio Gomes Lopes, Núbia Mendonça, Maria Lúcia M. Lee, Luís Fernando Lopes, C Mendonça, Flávia Aparecida Nogueira, Flávia Cristina Fernandes Pimenta, Vitória P. Pinheiro, Denise Bousfield da Silva, Elaine Sobral da Costa, Fernando Regla Vargas, Fernando de Almeida Werneck
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Abstract
Denaturing high-performance liquid chromatography (dHPLC) was developed to screen DNA variations by separating heteroduplex and homoduplex DNA fragments by ion-pair reverse-phase liquid chromatography. In this study, we have evaluated the dHPLC screening method and direct sequencing for the detection of GATA1 mutations in peripheral blood and bone marrow aspirates samples from children with Down syndrome (DS). Cases were ascertained consecutively as part of an epidemiological study of DS and hematological disorders in Brazil. A total of 130 samples corresponding to 115 children with DS were analysed using dHPLC and direct sequencing methods to detect mutations in GATA1 exons 2, 3 and 4 gene sequences. The overall detection rate of sequencing and dHPLC screening methods was similar. Twenty mutations were detected in exon 2 and one mutation in exon 3 (c.231_232 dupGT) sequences of acute megakaryoblastic leukemia and transient leukemia samples. Four GATA1 mutations were newly described [c.155C > G; c.156_178 del23 bp; c.29_30 del GG; c.182C > A and c.151A > T,c.153_162 del 10 bp). Out of four, three had single nucleotide change. In conclusion, our results indicate that dHPLC is an efficient and valuable tool for GATA1 mutational analysis.