Detection of mutations in GATA1 gene using automated denaturing high-performance liquid chromatography and direct sequencing in children with Down syndrome.
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.
Amorim
Marcia R
MR
Figueiredo
Alexandre B C
AB
Splendore
Alessandra
A
Magalhães
Isis Q
IQ
Pombo-de-Oliveira
Maria S
MS
El-Jaick
Kênia B
KB
D'andrea
Maria Lydia
ML
Aquino
Jozina
J
Alencar
Dora Márcia
DM
Brandalise
Silvia R
SR
Burlemaqui
Lilian
L
Cardoso
Teresa Cristina
TC
Carvalho
Eni Guimarães
EG
Coser
Virginia M
VM
Costa
Imaruí
I
Dorea
Dolores
D
Drumond
Mauricio
M
Lopes
Venâncio Gomes
VG
Mendonça
Núbia
N
Lee
Maria Lucia M
ML
Lopes
Luis Fernando
LF
Mendonça
Carmen M
CM
Nogueira
Flávia
F
Pimenta
Flávia
F
Pinheiro
Vitória P
VP
Da Silva
Denise Bousfield
DB
Sobral
Elaine
E
Vargas
Fernando R
FR
Werneck
Fernando
F
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
D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human