Amplification of X- and Y-Chromosome-Specific Regions from Single Human Blastomeres by Polymerase Chain Reaction

Fadel A Sharif, Amani N Azarah

Abstract


Objective: This study was conducted in order to investigate the feasibility of applying whole genomic DNA amplification (WGA) and polymerase chain reaction (PCR) techniques for sex determination of single human blastomere for the first time in Gaza Strip. Samples and Methods: In this study, WGA technique was employed on single blastomeres that were biopsied from 30 surplus human embryos donated for this research. The obtained genomic DNAs were then subjected for PCR amplification of the Y-linked SRY (Sex-Determining Region on Y chromosome) and the X-linked glucose-6-phosphate dehydrogenase (G6PD) genes. Results: Whole genomic DNA amplification efficiency from single blastomeres was 100 % and the level of amplification ranged from about 40 to 100 fold. All amplified DNA samples were included in the subsequent sex determination PCR analysis. Of the 30 embryos, 7 (23%) were found to be male and the remaining 23 embryos (77%) were of female gender. Conclusion: Based on these results, WGA is effective for obtaining sizable amounts of DNA from single blastomeres. Moreover, the obtained DNA proved amenable to PCR and this will pave the way for pre-implantation genetic diagnosis of various inherited disorders and for non-medical sex selection in Gaza strip.

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