Comparison of paired cerebrospinal fluid and serum cell-free mitochondrial and nuclear DNA with copy number and fragment length
Comparison of paired cerebrospinal fluid and serum cell-free mitochondrial and nuclear DNA with copy number and fragment length
复制标题
配对脑脊液和血清中游离线粒体和核 DNA 的拷贝数和片段长度的比较
DOI：
10.1002/jcla.23238
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发表时间：
2020-02-13
影响因子：
2.7
通讯作者：
Wu, Wenhe
中科院分区：
文献类型：
--
作者：
Chen, Aolong;Li, Jun;Wu, Wenhe
背景大多数无细胞DNA（CFDNA）的研究仅用于单人体流体。但是，很少报道两种体液之间CFDNA分布的差异。因此，在这项工作中，我们比较了脑脊液（CSF）与脑相关疾病患者的血清之间CFDNA分布的差异。方法使用Agilent 2100 Bioanalyer确定CfDNA的片段长度。通过使用实时定量PCR（QPCR）和液滴数字PCR（DDPCR）确定无细胞线粒体DNA（CF-MTDNA）和无细胞核DNA（CF-NDNA）的拷贝数。分别和核GAPDH底漆。在其中的较短（类似于60 bp），培养基（类似于167 bp）和长（> 250 bp）CFDNA片段长度分布完全从CSF和血清获得，使用Agilent 2100生物分析仪获得。 QPCR和DDPCR的结果证实了CSF和血清中这三个CFDNA片段范围的存在。根据QPCR，CSF中长CF-MTDNA，培养基和长CF-NDNA的拷贝数显着高于配对的血清中。在CSF中，只有长CF-MTDNA的拷贝数高于长CF-NDNA。但是在血清中，培养基和长CF-MTDNA的拷贝数高于相应的CF-NDNA。结论的CF-NDNA和CF-MTDNA具有不同片段长度的CF-NDNA和CF-MTDNA，在CSF中分布在CSF和脑疾病患者的血清中，这是脑疾病的患者的血清，可能是人脑疾病的生物标志物。
Background Most studies on cell-free DNA (cfDNA) were only for single body fluids; however, the differences in cfDNA distribution between two body fluids are rarely reported. Hence, in this work, we compared the differences in cfDNA distribution between cerebrospinal fluid (CSF) and serum of patients with brain-related diseases.Methods The fragment length of cfDNA was determined by using Agilent 2100 Bioanalyzer. The copy numbers of cell-free mitochondrial DNA (cf-mtDNA) and cell-free nuclear DNA (cf-nDNA) were determined by using real-time quantitative PCR (qPCR) and droplet digital PCR (ddPCR) with three pairs of mitochondrial ND1 and nuclear GAPDH primers, respectively.Results There were short (similar to 60 bp), medium (similar to 167 bp), and long (>250 bp) cfDNA fragment length distributions totally obtained from CSF and serum using Agilent 2100 Bioanalyzer. The results of both qPCR and ddPCR confirmed the existence of these three cfDNA fragment ranges in CSF and serum. According to qPCR, the copy numbers of long cf-mtDNA, medium, and long cf-nDNA in CSF were significantly higher than in paired serum. In CSF, only long cf-mtDNA's copy numbers were higher than long cf-nDNA. But in serum, the copy numbers of medium and long cf-mtDNA were higher than the corresponding cf-nDNA.Conclusion The cf-nDNA and cf-mtDNA with different fragment lengths differentially distributed in the CSF and serum of patients with brain disorders, which might serve as a biomarker of human brain diseases.