Preimplantation Genetic Diagnosis for β-Thalassemia Using Single-cell DNA Analysis for Codons 17 and 26 of β-globin Gene

Noor Wahidah Mohd Nasri, A. Rahman A. Jamal, Nurshaireen Chue Abdullah, Zainul Rashid Mohd. Razi, Norfilza Mohd Mokhtar

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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 β-globin gene that cause the β-thalassemia disorder. We developed a rapid and highly specific technique of single-cell PCR to amplify a specific region on the β-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. Crown

Original languageEnglish
Pages (from-to)1-9
Number of pages9
JournalArchives of Medical Research
Volume40
Issue number1
DOIs
Publication statusPublished - Jan 2009

Fingerprint

Single-Cell Analysis
Preimplantation Diagnosis
Thalassemia
Globins
Codon
Blastomeres
Polymerase Chain Reaction
DNA
Inborn Genetic Diseases
Embryonic Structures
Genes
Blastocyst
Genome
Spontaneous Abortion
DNA-Directed DNA Polymerase
Crowns
Prenatal Diagnosis
Pregnancy
Mutation

Keywords

  • β-Thalassemia
  • Genetic
  • Preimplantation diagnosis
  • Whole genome amplification

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Preimplantation Genetic Diagnosis for β-Thalassemia Using Single-cell DNA Analysis for Codons 17 and 26 of β-globin Gene. / Mohd Nasri, Noor Wahidah; A. Jamal, A. Rahman; Abdullah, Nurshaireen Chue; Mohd. Razi, Zainul Rashid; Mohd Mokhtar, Norfilza.

In: Archives of Medical Research, Vol. 40, No. 1, 01.2009, p. 1-9.

Research output: Contribution to journalArticle

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