血小板污染会导致外周血单个核细胞线粒体DNA含量出现较大差异以及高估。
Platelet contamination causes large variation as well as overestimation of mitochondrial DNA content of peripheral blood mononuclear cells.
作者信息
Urata Michiyo, Koga-Wada Yui, Kayamori Yuzo, Kang Dongchon
机构信息
Department of Clinical Chemistry and Laboratory Medicine, Kyushu University Graduate School of Medical Sciences, 3-1-1 Maidashi Higashi-ku, Fukuoka, Japan.
出版信息
Ann Clin Biochem. 2008 Sep;45(Pt 5):513-4. doi: 10.1258/acb.2008.008008.
BACKGROUND
Alterations in the copy number of mitochondrial DNA (mtDNA) play a role in the pathogenesis of mitochondrial diseases and other many common diseases. Recently, the copy number of leukocyte mtDNA has been considered to serve as a biomarker to monitor or chase such diseases. Therefore, reproducible mtDNA measurement is required.
METHODS
Peripheral blood mononuclear cells were prepared by a density-based method. The mtDNA/cell was measured by quantitative realtime polymerase chain reaction.
RESULTS
The degree of platelet contamination varied to a large extent among preparations. The mtDNA copy numbers per mononuclear cell were 269 +/- 51 and 146 +/- 14 in the samples before and after the platelet depletion, respectively.
CONCLUSION
A density-based mononuclear cell preparation causes heavy platelet contamination. The platelet depletion from a sample is particularly important for comparing the mtDNA contents between different dates or between different patients.
背景
线粒体DNA（mtDNA）拷贝数的改变在线粒体疾病及其他多种常见疾病的发病机制中起作用。近来，白细胞mtDNA拷贝数被认为可作为监测或追踪此类疾病的生物标志物。因此，需要可重复的mtDNA测量方法。
方法
采用基于密度的方法制备外周血单个核细胞。通过定量实时聚合酶链反应测量每个细胞的mtDNA。
结果
不同制备物中血小板污染程度差异很大。血小板去除前后，样本中每个单核细胞的mtDNA拷贝数分别为269±51和146±14。
结论
基于密度的单核细胞制备会导致严重的血小板污染。从样本中去除血小板对于比较不同时间或不同患者之间的mtDNA含量尤为重要。
Zotero 插件