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

Michiyo Urata, Machiko Wakiyama, Masanori Iwase, Makoto Yoneda, Sachiko Kinoshita, Naotaka Hamasaki, Dongchon Kang

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)2088-2093
Number of pages6
JournalClinical Chemistry
Volume44
Issue number10
Publication statusPublished - Oct 12 1998

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Polymerase chain reaction
Medical problems
Ligation
Diabetes Mellitus
Blood
Nucleotides
RNA, Transfer, Leu
Cells
Polymerase Chain Reaction
Mutation
DNA
Guanine
Adenine
Genes
Tissue
Blood Cells
Enzymes
Leukocytes
Phenotype

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

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. / Urata, Michiyo; Wakiyama, Machiko; Iwase, Masanori; Yoneda, Makoto; Kinoshita, Sachiko; Hamasaki, Naotaka; Kang, Dongchon.

In: Clinical Chemistry, Vol. 44, No. 10, 12.10.1998, p. 2088-2093.

Research output: Contribution to journalArticle

Urata, Michiyo ; Wakiyama, Machiko ; Iwase, Masanori ; Yoneda, Makoto ; Kinoshita, Sachiko ; Hamasaki, Naotaka ; Kang, Dongchon. / 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. In: Clinical Chemistry. 1998 ; Vol. 44, No. 10. pp. 2088-2093.
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