Detection of mitochondrial DNA deletion by a modified PCR method in a 60Co radiation-exposed patient.
A new PCR based method was developed to detect deleted mitochondrial DNA (mtDNA). Peripheral blood cell DNA was obtained from a victim who was accidently exposed to a 60Co radiation source in 1990. Using the DNA as template, first PCR was performed to generate multiple products including true deletions and artifacts. The full length product was recovered and used as template of secondary PCR. The suspicious deletion product of mtDNA could be confirmed only if it was yielded by first PCR. Using either original primers or their nested primers, the suspicious deletion product was amplified and authenticated as a true deletion product. The template was recovered and determined to be a deletion by sequencing directly. The results show that a new mtDNA deletion, which spans 889 bp from nt 11688 to nt 12576, was detected in the peripheral blood cells of the victim. It indicates that this new PCR-based method was more efficient at detecting small populations of mtDNA deletion than other routine methods. MtDNA deletion was found in the victim, suggesting the relationship between the deletion and phenotypes of the disease.
Wang
Zhen-Cheng
ZC
Department of Basic Medicine, Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai, 200433, People's Republic of China.
Wang
Xue-Min
XM
Jiao
Bing-Hua
BH
Jin
You-Xin
YX
Miao
Ming-Yong
MY
Zhu
Ke-Jun
KJ
Ni
Qing-Gui
QG
D000328:Adult; D001483:Base Sequence; D003037:Cobalt Radioisotopes / Q000009:adverse effects*; D004247:DNA / Q000097:blood; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000528:radiation effects*; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D018788:Radioactive Hazard Release; D017384:Sequence Deletion
D003037:Cobalt Radioisotopes; D004272:DNA, Mitochondrial; D004247:DNA