Profile of the Circulating DNA in Apparently Healthy Individuals
Profile of the Circulating DNA in Apparently Healthy Individuals
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DOI：
10.1373/clinchem.2008.113597
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发表时间：
2009-04-01
影响因子：
9.3
通讯作者：
Schuetz, Ekkehard
中科院分区：
文献类型：
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作者：
Beck, Julia;Urnovitz, Howard B.;Schuetz, Ekkehard
背景：循环核酸（CNA）已被证明在人类疾病中具有诊断效用。本研究的目的是对CNA进行测序和组织，以记录表面健康个体循环DNA的典型特征。方法：从51名表面健康的人中提取血清DNA，扩增，通过焦磷酸测序进行测序（454 Life Sciences/Roche Diagnostics），并按（a）来源分类（人与异种），（B）功能性（重复，基因，编码或非编码），和（c）染色体定位。CNA结果与基因组DNA对照（n = 4）进行了比较，进行了相同的proceeding.RESULTS：我们获得了4.5 × 10（5）序列（7.5 × 10（7）核苷酸），其中87%是归因于已知的数据库序列。在这些序列中，97%是基因组的，3%是异种的。CNA和基因组DNA没有不同的重复序列，基因，RNA和蛋白质编码的DNA序列。CNA中短散在核元件序列所占比例较高（P = 0.1），其中Alu序列所占比例显著（P < 0.01）。CNAs中L1和L2长散布核元件序列所占比例显著低于对照组（P < 0.01）。此外，B型肝炎病毒（HBV）基因型F序列被发现在一个人意外评估作为一个健康的control.CONCLUSIONS：CNAs与基因组DNA的比较表明，非特异性DNA释放是不是唯一的起源CNAs。我们所描述的健康个体的CNA分析，以及详细的生物特征分析，为未来对特定疾病患者的研究提供了基础。此外，以前未知的HBV感染的检测表明该方法能够发现隐匿性感染。(C)2009年美国临床化学协会
BACKGROUND: Circulating nucleic acids (CNAs) have been shown to have diagnostic utility in human diseases. The aim of this study was to sequence and organize CNAs to document typical profiles of circulating DNA in apparently healthy individuals.METHODS: Serum DNA from 51 apparently healthy humans was extracted, amplified, sequenced via pyro-sequencing (454 Life Sciences/Roche Diagnostics), and categorized by (a) origin (human vs xenogeneic), (b) functionality (repeats, genes, coding or noncoding), and (c) chromosomal localization. CNA results were compared with genomic DNA controls (n = 4) that were subjected to the identical procedure.RESULTS: We obtained 4.5 x 10(5) sequences (7.5 x 10(7) nucleotides), of which 87% were attributable to known database sequences. Of these sequences, 97% were genomic, and 3% were xenogeneic. CNAs and genomic DNA did not differ with respect to sequences attributable to repeats, genes, RNA, and protein-coding DNA sequences. CNA tended to have a higher proportion of short interspersed nuclear element sequences (P = 0.1), of which Alu sequences were significant (P < 0.01). CNAs had a significantly lower proportion of L1 and L2 long interspersed nuclear element sequences (P < 0.01). In addition, hepatitis B virus (HBV) genotype F sequences were found in an individual accidentally evaluated as a healthy control.CONCLUSIONS: Comparison of CNAs with genomic DNA suggests that nonspecific DNA release is not the sole origin for CNAs. The CNA profiling of healthy individuals we have described, together with the detailed biometric analysis, provides the basis for future studies of patients with specific diseases. Furthermore, the detection of previously unknown HBV infection suggests the capability of this method to uncover occult infections. (C) 2009 American Association for Clinical Chemistry