TY - JOUR
T1 - New sensitive method for the detection of the A3243G mutation of human mitochondrial deoxyribonucleic acid in diabetes mellitus patients by ligation-mediated polymerase chain reaction
AU - Urata, Michiyo
AU - Wakiyama, Machiko
AU - Iwase, Masanori
AU - Yoneda, Makoto
AU - Kinoshita, Sachiko
AU - Hamasaki, Naotaka
AU - Kang, Dongchon
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1998
Y1 - 1998
N2 - An adenine-to-guanine mutation at nucleotide position (np) 3243 in the mitochondrial tRNALeu(UUR) gene is closely associated with various clinical phenotypes of diabetes mellitus. Because the mutation creates a new restriction site for the restriction enzyme ApaI, the mutation is usually detected and quantified by ApaI cleavage of the PCR products including np 3243. The sensitivity of the conventional method is, however, 5-10% heteroplasmy. The percentage of heteroplasmy of the mutation is usually highest in the affected tissues and is much lower in peripheral blood cells, which are used most frequently for the analysis. The sensitivity of the conventional method, however, is not sufficient to detect the mutation from peripheral blood cells. Utilizing ligation-mediated polymerase chain reaction, we have developed a feasible and sensitive method to detect 0.01% heteroplasmy of the 3243 mutation in peripheral leukocytes.
AB - An adenine-to-guanine mutation at nucleotide position (np) 3243 in the mitochondrial tRNALeu(UUR) gene is closely associated with various clinical phenotypes of diabetes mellitus. Because the mutation creates a new restriction site for the restriction enzyme ApaI, the mutation is usually detected and quantified by ApaI cleavage of the PCR products including np 3243. The sensitivity of the conventional method is, however, 5-10% heteroplasmy. The percentage of heteroplasmy of the mutation is usually highest in the affected tissues and is much lower in peripheral blood cells, which are used most frequently for the analysis. The sensitivity of the conventional method, however, is not sufficient to detect the mutation from peripheral blood cells. Utilizing ligation-mediated polymerase chain reaction, we have developed a feasible and sensitive method to detect 0.01% heteroplasmy of the 3243 mutation in peripheral leukocytes.
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U2 - 10.1093/clinchem/44.10.2088
DO - 10.1093/clinchem/44.10.2088
M3 - Article
C2 - 9761239
AN - SCOPUS:0031664644
VL - 44
SP - 2088
EP - 2093
JO - Clinical Chemistry
JF - Clinical Chemistry
SN - 0009-9147
IS - 10
ER -