Genetic screening of Leber’s hereditary optic neuropathy by PCR with whole blood cell lysate

Abstract

Leber’s hereditary optic neuropathy (LHON) is accompanied by a mitochondrial DNA (mtDNA) mutation. The G to A substitution at nucleotide position 11,778 (11,202) of mtDNA is most common in Japanese LHON patients. Whole blood cell lysate, not purified DNA, was used as a template of the polymerase chain reaction (PCR) for the analysis of the G11,778 (11,202)A point mutation. The amplified DNA fragment was concentrated and desalted with a centrifuge device, SUPREC TM -02, and digested by SfaNI and MaeIII. This method does not need purified DNA from blood and avoids the phenol/chloroform treatments for PCR products prior to the restriction enzyme digestion. Therefore, it is convenient and safe for the genetic screening of LHON.