Preimplantation genetic diagnosis for beta-thalassemia using single-cell DNA analysis for codons 17 and 26 of beta-globin gene.

BACKGROUND: Preimplantation genetic diagnosis (PGD) of monogenic autosomal hereditary disorders following assisted conception usually involves the removal of one or two blastomeres from preimplantation embryos. However, the amount of DNA from a single blastomere is insufficient to amplify the region of interest. Hence, the whole genome amplification (WGA) method is performed prior to amplifying the genes of interest before analysis of DNA material through polymerase chain reaction (PCR). METHODS: In the present study we report that WGA from a single blastomere extracted from unwanted preimplantation human embryos (obtained from 10 infertile couples) could positively yield microgram quantities of amplified DNA allowing PCR analysis for codons 17 and 26 of the beta-globin gene that cause the beta-thalassemia disorder. We developed a rapid and highly specific technique of single-cell PCR to amplify a specific region on the beta-globin gene for codon 17 (AAG-->TAG) and codon 26 (GAG-->AAG) by using single-cell PCR. RESULTS: About 249 bp of amplicon for codon 17 and about 200 bp of amplicon for codon 26 were successfully amplified. No mutations were observed. Analyzed embryos were not transferred back to patients because the embryos used as samples were wasted embryos. CONCLUSIONS: Compared to other approaches for prenatal diagnosis, PGD is rapid and suitable as a noninvasive clinical tool for identifying genetic disorders for the purpose of reducing selective miscarriages and moral dilemmas. We opine that DNA extraction and amplification can be successfully performed by using single-cell PCR to diagnose genetic diseases before pregnancy.

Dengue in pregnancy.

This was a retrospective study of patients with dengue infection in pregnancy from year 2000 till 2004. Data were analyzed by looking at the presentation, complications of patient and fetus, and pregnancy outcomes. There was a total of 16 cases with an increasing trend (0.12% in 2003 vs 0.25% in 2004). The mean age of patients was 30.19+/-6.85 years. Fifty percent of patients were multiparae and in their third trimester. The average gestation was 24.44 weeks with 7.5 days being the average duration of ward admission. Tourniquet test was positive in 62.5% of patients. Dengue serology IgM was positive in 50% whereas dengue serology IgG were positive in 68.8% of patients. There were three cases of maternal death. One patient presented as missed abortion. Preterm deliveries happened in 50.0% of the women. There were 4 premature babies, one in-utero fetal death, and two fetuses which suffered acute fetal distress. Three babies required intensive care. One unrelated fetal anomaly resulted in early neonatal death.