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Preimplantation genetic diagnosis for beta-thalassemia using single-cell DNA analysis for codons 17 and 26 of beta-globin gene.
Arch Med Res 2009; 40(1):1-9AM

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 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.

Authors+Show Affiliations

Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19064120

Citation

Nasri, Noor Wahidah Mohd, et al. "Preimplantation Genetic Diagnosis for Beta-thalassemia Using Single-cell DNA Analysis for Codons 17 and 26 of Beta-globin Gene." Archives of Medical Research, vol. 40, no. 1, 2009, pp. 1-9.
Nasri NW, Jamal AR, Abdullah NC, et al. Preimplantation genetic diagnosis for beta-thalassemia using single-cell DNA analysis for codons 17 and 26 of beta-globin gene. Arch Med Res. 2009;40(1):1-9.
Nasri, N. W., Jamal, A. R., Abdullah, N. C., Razi, Z. R., & Mokhtar, N. M. (2009). Preimplantation genetic diagnosis for beta-thalassemia using single-cell DNA analysis for codons 17 and 26 of beta-globin gene. Archives of Medical Research, 40(1), pp. 1-9. doi:10.1016/j.arcmed.2008.10.008.
Nasri NW, et al. Preimplantation Genetic Diagnosis for Beta-thalassemia Using Single-cell DNA Analysis for Codons 17 and 26 of Beta-globin Gene. Arch Med Res. 2009;40(1):1-9. PubMed PMID: 19064120.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Preimplantation genetic diagnosis for beta-thalassemia using single-cell DNA analysis for codons 17 and 26 of beta-globin gene. AU - Nasri,Noor Wahidah Mohd, AU - Jamal,A Rahman A, AU - Abdullah,Nurshaireen Chue, AU - Razi,Zainul Rashid Mohd, AU - Mokhtar,Norfilza Mohd, PY - 2008/04/24/received PY - 2008/10/06/accepted PY - 2008/12/10/pubmed PY - 2009/2/13/medline PY - 2008/12/10/entrez SP - 1 EP - 9 JF - Archives of medical research JO - Arch. Med. Res. VL - 40 IS - 1 N2 - 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. SN - 1873-5487 UR - https://www.unboundmedicine.com/medline/citation/19064120/Preimplantation_genetic_diagnosis_for_beta_thalassemia_using_single_cell_DNA_analysis_for_codons_17_and_26_of_beta_globin_gene_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0188-4409(08)00253-1 DB - PRIME DP - Unbound Medicine ER -