title: Detection and quantification of the age-related point mutation A189G in the human mitochondrial DNA.
abstract: Mutation analysis in the mitochondrial DNA (mtDNA) control region is widely used in population genetic studies as well as in forensic medicine. Among the difficulties linked to the mtDNA analysis, one can find the detection of heteroplasmy, which can be inherited or somatic. Recently, age-related point mutation A189G was described in mtDNA and shown to accumulate with age in muscles. We carried out the detection of this 189 heteroplasmic point mutation using three technologies: automated DNA sequencing, Southern blot hybridization using a digoxigenin-labeled oligonucleotide probe, and peptide nucleic acid (PNA)/real-time PCR combined method on different biological samples. Our results give additional information on the increase in mutation frequency with age in muscle tissue and revealed that the PNA/real-time PCR is a largely more sensitive method than DNA sequencing for heteroplasmy detection. These investigations could be of interest in the detection and interpretation of mtDNA heteroplasmy in anthropological and forensic studies.
authors.affiliation: INSERM, U563 Bat C, Purpan University Hospital, Place du Dr Baylac, 31059 Toulouse, France. Catherine.Theves@toulouse.inserm.fr
authors.forename: Catherine
authors.initials: C
authors.lastname: Thèves
authors.forename: Christine
authors.initials: C
authors.lastname: Keyser-Tracqui
authors.forename: Eric
authors.initials: E
authors.lastname: Crubézy
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Salles
authors.forename: Bertrand
authors.initials: B
authors.lastname: Ludes
authors.forename: Norbert
authors.initials: N
authors.lastname: Telmon
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D015342:DNA Probes; D004272:DNA, Mitochondrial / Q000235:genetics*; D006801:Humans; D007223:Infant; D008875:Middle Aged; D009061:Mouth Mucosa / Q000166:cytology; D020135:Peptide Nucleic Acids; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D016658:Psoas Muscles / Q000166:cytology; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004272:DNA, Mitochondrial; D020135:Peptide Nucleic Acids