title: Detection of mutations in GATA1 gene using automated denaturing high-performance liquid chromatography and direct sequencing in children with Down syndrome.
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.
authors.forename: Marcia R
authors.initials: MR
authors.lastname: Amorim
authors.forename: Alexandre B C
authors.initials: AB
authors.lastname: Figueiredo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Splendore
authors.forename: Isis Q
authors.initials: IQ
authors.lastname: Magalhães
authors.forename: Maria S
authors.initials: MS
authors.lastname: Pombo-de-Oliveira
authors.forename: Kênia B
authors.initials: KB
authors.lastname: El-Jaick
authors.forename: Maria Lydia
authors.initials: ML
authors.lastname: D'andrea
authors.forename: Jozina
authors.initials: J
authors.lastname: Aquino
authors.forename: Dora Márcia
authors.initials: DM
authors.lastname: Alencar
authors.forename: Silvia R
authors.initials: SR
authors.lastname: Brandalise
authors.forename: Lilian
authors.initials: L
authors.lastname: Burlemaqui
authors.forename: Teresa Cristina
authors.initials: TC
authors.lastname: Cardoso
authors.forename: Eni Guimarães
authors.initials: EG
authors.lastname: Carvalho
authors.forename: Virginia M
authors.initials: VM
authors.lastname: Coser
authors.forename: Imaruí
authors.initials: I
authors.lastname: Costa
authors.forename: Dolores
authors.initials: D
authors.lastname: Dorea
authors.forename: Mauricio
authors.initials: M
authors.lastname: Drumond
authors.forename: Venâncio Gomes
authors.initials: VG
authors.lastname: Lopes
authors.forename: Núbia
authors.initials: N
authors.lastname: Mendonça
authors.forename: Maria Lucia M
authors.initials: ML
authors.lastname: Lee
authors.forename: Luis Fernando
authors.initials: LF
authors.lastname: Lopes
authors.forename: Carmen M
authors.initials: CM
authors.lastname: Mendonça
authors.forename: Flávia
authors.initials: F
authors.lastname: Nogueira
authors.forename: Flávia
authors.initials: F
authors.lastname: Pimenta
authors.forename: Vitória P
authors.initials: VP
authors.lastname: Pinheiro
authors.forename: Denise Bousfield
authors.initials: DB
authors.lastname: Da Silva
authors.forename: Elaine
authors.initials: E
authors.lastname: Sobral
authors.forename: Fernando R
authors.initials: FR
authors.lastname: Vargas
authors.forename: Fernando
authors.initials: F
authors.lastname: Werneck
mesh_terms: D001938:Brazil / Q000453:epidemiology; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D004252:DNA Mutational Analysis / Q000379:methods*; D004314:Down Syndrome / Q000150:complications / Q000453:epidemiology / Q000235:genetics*; D005260:Female; D050982:GATA1 Transcription Factor / Q000235:genetics*; D006402:Hematologic Diseases; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007938:Leukemia / Q000235:genetics; D008297:Male; D009154:Mutation; D017422:Sequence Analysis, DNA
chemical_list: D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human