Prenatal Diagnosis of Homozygous Beta-Thalassemia by Fetal Hb Typing Analysis Using Automated HPLC
Keywords:
PND, Homozygous B-thalassemia, Fetal Hb typing, HPLCAbstract
Background: Prenatal diagnosis is one of the method used for prevention and control of homozygous B-thalassemia. The method usually used is DNA analysis from chorionic vili and amniocytes, which are quite complicated and expensive. However, in Thailand the causes of this disease are mostly due to to -thalassemia genes, which cannot produce Hb A. Therefore, the measurement of Hb A percentage by automated HPLC could be an other helpful prenatal diagnostic method. Materials and methods: Forty pregnant
women, at a gestational age of between 18:22 weels were chosen. They were at risk of delivening a child with homozygous B-thalassemia. Cordocentesis was performed and fetal blood analyzed for Hb typing by automated HPLC. The P-globin gene mutation was demonstrated by direct B-globin gene sequencing. Results: There were 9 fetuses which had Hb F only. The DNA sequencing analysis of them showed 2 fetuses were homozygote of 4 bp deletion at codon 41/42 (-CTTT), 4 were combound heterozygoteof non-
sense mutation at codon 17 (A-T) and 4 bp deletion at codon 41/42 (-CTTT), 2 were homozygote of splice site mutation at IVSinti(G-T) and one fetus was compound heterozygote of nonsense mutation at codon 35(C-A) and 4 bo deletion at codon 41/42 (-CTTT). Thirty one fetuses showed Hb A in the HPLCchromatograms with the proportion from 05 to 7.4%. One fetus with Hb A of 05% was a case of compoundheterozygous promoter mutation at -28 (A-G) and 4bp deletion of codon 41/42 (-CTT). Twenty fetuses were B-thalassemia trait where 12 were 4 bp deletion at codon 41/42 (-CTT), 3 were IVSinti(G-T), 2 were nonsense mutation of codon 35 (C-A), 2 were promoter mutation at position -28 (A-G) and 1 was nonsensemutation at codon 17 (A-T). The proportion of Hb A of this group was 08-2.8% (1.8220.49). The other 10 fetuses showed Hb A of 2.9 - 7.4% (4.921.5) that had normal DNA sequencing. Conclusion: Prenatal diagnosis of homozygous B-thalassemia by Hb typing analysis using automated HPLC showed an absence of Hb A, which was the characteristic finding. The results were confimed by genotypes from DNA sequencing. It was suggested that the analysis of fetal hemoglobin is simple inexpensive and accurate for prenatal diagnosis. Therefore this method would be the most appropriate for the prevention and control of homozygous B-thalassemia throughout Thailand.
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