A rapid screening with direct sequencing from blood samples for the diagnosis of Leigh syndrome

Hiroko Shimbo; Mariko Takagi; Mitsuko Okuda; Yu Tsuyusaki; Kyoko Takano; Mizue Iai; Sumimasa Yamashita; Kei Murayama; Akira Ohtake; Yu-Ichi Goto; Noriko Aida; Hitoshi Osaka

doi:10.1016/j.ymgmr.2014.02.006

A rapid screening with direct sequencing from blood samples for the diagnosis of Leigh syndrome

Mol Genet Metab Rep. 2014 Apr 1:1:133-138. doi: 10.1016/j.ymgmr.2014.02.006. eCollection 2014.

Authors

Affiliations

¹ Division of Neurology, Kanagawa Children's Medical Center, 2-138-4 Mutsukawa, Minami-ku, Yokohama, Kanagawa 232-8555, Japan.
² Department of Metabolism, Chiba Children's Hospital, 579-1, Heta-cho, Midori-ku, Chiba-shi, Chiba 266-0007, Japan.
³ Department of Pediatrics, Faculty of Medicine, Saitama Medical University, 38 Morohongo, Moroyama, Iruma-gun, Saitama 350-0495, Japan.
⁴ Department of Mental Retardation and Birth Defect Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 4-1-1 Ogawahigashi-machi, Kodaira-shi, Tokyo 187-8551, Japan.
⁵ Division of Radiology, Kanagawa Children's Medical Center, 2-138-4 Mutsukawa, Minami-ku, Yokohama, Kanagawa 232-8555, Japan.
⁶ Division of Neurology, Kanagawa Children's Medical Center, 2-138-4 Mutsukawa, Minami-ku, Yokohama, Kanagawa 232-8555, Japan; Department of Pediatrics, Jichi Medical School, 3311-1 Yakushiji, Shimotsuke-shi, Tochigi 329-0498, Japan.

Abstract

Large numbers of genes are responsible for Leigh syndrome (LS), making genetic confirmation of LS difficult. We screened our patients with LS using a limited set of 21 primers encompassing the frequently reported gene for the respiratory chain complexes I (ND1-ND6, and ND4L), IV(SURF1), and V(ATP6) and the pyruvate dehydrogenase E1α-subunit. Of 18 LS patients, we identified mutations in 11 patients, including 7 in mDNA (two with ATP6), 4 in nuclear (three with SURF1). Overall, we identified mutations in 61% of LS patients (11/18 individuals) in this cohort. Sanger sequencing with our limited set of primers allowed us a rapid genetic confirmation of more than half of the LS patients and it appears to be efficient as a primary genetic screening in this cohort.

Keywords: Complex I deficiency; Heteroplasmy; Leigh syndrome; mDNA mutation.