中林　章    Department   School of Medicine(Tokyo Women's Medical University Hospital), School of Medicine    Position   Associate Professor
Article types	Original article
Language	English
Peer review	Peer reviewed
Title	The development of novel quantification assay for mitochondrial DNA heteroplasmy aimed at preimplantation genetic diagnosis of Leigh encephalopathy.
Journal	Formal name：Journal of assisted reproduction and genetics Abbreviation：J Assist Reprod Genet ISSN code：10580468/10580468
Domestic / Foregin	Foregin
Volume, Issue, Page	24(6),pp.227-32
Author and coauthor	Tajima Hiroto, Sueoka Kou, Moon Sung Yung, Nakabayashi Akira, Sakurai Tomoyoshi, Murakoshi Yukitaka, Watanabe Hiroyoshi, Iwata Soukichi, Hashiba Tsuyoshi, Kato Shingo, Goto Yu-Ichi, Yoshimura Yasunori
Publication date	2007/06
Summary	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.
DOI	10.1007/s10815-007-9114-0
PMID	17342424