Quantification of mitochondrial DNA in human blood cells using an automated detection system.
The 4977 bp deletion of mitochondrial DNA (mtDNA) accumulates in postmitotic tissues with advancing age. The purpose of our study was to detect and quantify these deletion even in blood cells with a high turnover activity. Whole venous blood, isolated human platelets and peripheral blood mononuclear cells (PBMCs) were collected from 10 unrelated donors aged 20-71 years and total DNA was extracted. PCR was performed for total and mutated mtDNA using two different primer pairs and two fluorogenic probes labeled with the fluorescent dyes FAM and VIC. Specific PCR products were generated, detected and quantified in a real-time PCR. The amplification products of total and deleted mtDNA could be detected in each sample and did not exhibit any differences in the amount of the deleted mtDNA in whole blood, human platelets or PBMCs. Our data did not show any accumulation of the 4977 bp deletion with increasing age as it was observed for several other tissues.
Meissner
C
C
Department of Legal Medicine, Medical University of Luebeck, Kahlhorststrasse 31-35, 23562, Luebeck, Germany. meissner@rmed.mu-luebeck.de
Mohamed
S A
SA
Klueter
H
H
Hamann
K
K
von Wurmb
N
N
Oehmichen
M
M
D000328:Adult; D000368:Aged; D000375:Aging / Q000235:genetics*; D001483:Base Sequence / Q000235:genetics*; D001774:Blood Chemical Analysis / Q000295:instrumentation* / Q000379:methods*; D001792:Blood Platelets / Q000737:chemistry*; D004272:DNA, Mitochondrial / Q000032:analysis*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012044:Regression Analysis; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods*; D017384:Sequence Deletion / Q000235:genetics*
D004272:DNA, Mitochondrial