NEW SENSITIVE METHOD FOR THE DETECTION OF THE A3243G MUTATION OF HUMAN MITOCHONDRIAL DEOXYRIBONUCLEIC-ACID IN DIABETES-MELLITUS PATIENTS BYLIGATION-MEDIATED POLYMERASE-CHAIN-REACTION

Citation
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
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Medical Laboratory Technology
Journal title
ISSN journal
0009-9147
Volume
44
Issue
10
Year of publication
1998
Pages
2088 - 2093
Database
ISI
SICI code
0009-9147(1998)44:10<2088:NSMFTD>2.0.ZU;2-1
Abstract
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.