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The development of novel quantification assay for mitochondrial DNA heteroplasmy aimed at preimplantation genetic diagnosis of Leigh encephalopathy

  • Genetics
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Abstract

Purpose: To perform preimplantation genetic diagnosis (PGD) of Leigh encephalopathy, we developed a rapid and reliable quantification assay for the percentage of T8993G mtDNA mutation and analyzed various specimens. Methods: We prepared the standard curve by measuring serial proportion of 8993T/G cloned plasmid DNA using real-time PCR, and measured (1) mutant DNA (known proportions by PCR-RFLP), (2) single lymphocytes from 46% mutant carrier, (3) 123 blastomeres from 20 abnormal embryos. Results: (1) These were within −5∼+6% error range, (2) mean 44.3%(11–70%), (3) Five embryos harbored T8993G mutation (4–22%). Embryos from same person indicated different degrees of heteroplasmy, and blastomeres from same embryo demonstrated limited dispersion of heteroplasmy (2–11%). Conclusions: (1) This method provides rapid and reliable PGD for Leigh encephalopathy. (2) The variable heteroplasmy with somatic mitosis was suggested. (3) T8993G mutation was existed in undeveloped embryo, and the bottleneck theory was supported. The limited heteroplasmy dispersion of blastomeres from same embryo also supported reliability of PGD for T8993G mutation.

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Acknowledgments

I appreciate my coworkers’ collaborations and advice on the study, especially Ms. Yoko Yasuda, Ms. Maya Higuchi, Ms. Satoko Moriya, and Ms. Yuko Matumoto, and also alongside the financial and institutional support from the Department of Obstetrics and Gynecology, Keio University School of Medicine.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan

    Hiroto Tajima, Kou Sueoka, Sung Yung Moon, Akira Nakabayashi, Tomoyoshi Sakurai, Yukitaka Murakoshi, Hiroyoshi Watanabe, Soukichi Iwata, Tsuyoshi Hashiba & Yasunori Yoshimura

  2. Department of Microbiology and Immunology, Keio University School of Medicine, Tokyo, Japan

    Shingo Kato

  3. Department of Mental Retardation and Birth Defect Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan

    Yu-Ichi Goto

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  1. Hiroto Tajima
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  2. Kou Sueoka
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  3. Sung Yung Moon
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  4. Akira Nakabayashi
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  5. Tomoyoshi Sakurai
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  9. Tsuyoshi Hashiba
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  10. Shingo Kato
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  11. Yu-Ichi Goto
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  12. Yasunori Yoshimura
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Correspondence to Hiroto Tajima.

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Tajima, H., Sueoka, K., Moon, S.Y. et al. The development of novel quantification assay for mitochondrial DNA heteroplasmy aimed at preimplantation genetic diagnosis of Leigh encephalopathy. J Assist Reprod Genet 24, 227–232 (2007). https://doi.org/10.1007/s10815-007-9114-0

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  • DOI: https://doi.org/10.1007/s10815-007-9114-0

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