NEW SENSITIVE METHOD FOR THE DETECTION OF THE A3243G MUTATION OF HUMAN MITOCHONDRIAL DEOXYRIBONUCLEIC-ACID IN DIABETES-MELLITUS PATIENTS BYLIGATION-MEDIATED POLYMERASE-CHAIN-REACTION
M. Urata et al., NEW SENSITIVE METHOD FOR THE DETECTION OF THE A3243G MUTATION OF HUMAN MITOCHONDRIAL DEOXYRIBONUCLEIC-ACID IN DIABETES-MELLITUS PATIENTS BYLIGATION-MEDIATED POLYMERASE-CHAIN-REACTION, Clinical chemistry, 44(10), 1998, pp. 2088-2093
An adenine-to-guanine mutation at nucleotide position (np) 3243 in the
mitochondrial tRNALeu(UUR) gene is closely associated with various cl
inical 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, howe
ver, 5-10% heteroplasmy. The percentage of heteroplasmy of the mutatio
n is usually highest in the affected tissues and is much lower in peri
pheral blood cells, which are used most frequently for the analysis, T
he 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.