Abstract
A number of studies have shown that plasma cell-free DNA is closely related to the risk of stroke, but the fragmentation status of plasma cell-free DNA and its clinical application value in ischemic stroke are still unclear. In this study, 48 patients with new ischemic stroke and 20 healthy subjects were enrolled. The second-generation high-throughput sequencing technique was used to study the plasma cell-free fragment length and regional distribution of the subjects. As noted in our results, the ratio of plasma cell-free DNA fragments in the disease group was significantly greater than that of the healthy group in the 300–400 bp range; conversely for fragments at the 75–250 bp range, the ratio of plasma cell-free DNA fragments in the patient group was apparently lower than that of the healthy group. In-depth analysis of the proportion of fragments distributed on each component of the genome was carried out. Our results recorded that the plasma cell-free DNA fragments in the disease group were inclined to the EXON, CpG islands, and ALU regions in contrast to that of the healthy group. In particular, fragments within the 300–400 bp range of the disease group were enrichment in the regions of EXON, INTRON, INTERGENIC, LINE, Fragile, ALU, and CpG islands. In summary, our findings suggested that the intracellular DNA degradation profiles could be applied to distinguish the stroke group and the healthy group, which provided a theoretical basis for the clinical diagnosis and prognosis of stroke by profiling the characteristic of plasma cell-free DNA fragments.
Highlights
Stroke is an acute cerebrovascular disease characterized by focal neurological deficits
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. e study had been approved by the Ethics Review Committee in Jining Medical University. e cell-free DNA of plasma samples were subsequently subjected to library construction and sequencing by the BGISEQ-500 sequencer according to the BGISEQ-500 protocol. e BWA algorithm was used to align the reads to the human reference genome hg19, and duplications were removed according to the previous study [19, 20]. e subsequent analysis was performed by an in-house developed bioinformatic pipeline
Cell-Free DNA Fragment Size Distribution and Concentration in the Plasma. e length distribution of plasma cellfree DNA fragments revealed the average fragment ratio of specific fragment size in two groups. It showed that the proportion of plasma cell-free DNA fragments (75–250 bp) in the healthy group was significantly greater than that of the ischemic stroke (IS) patients (Figure 1(a))
Summary
Stroke is an acute cerebrovascular disease characterized by focal neurological deficits. Several other studies revealed that there were different characteristics of the length of plasma cell-free DNA fragments and genomic regional distribution in different diseases [12, 17, 18]. The fragmentation state of plasma cell-free DNA in stroke patients and its association with clinical studies remain unclear. Deploying a secondgeneration high-throughput sequencing platform, our team had profiled the length and genomic regional distribution of cell-free DNA fragments in the healthy individuals and the ischemic stroke patient. This research had compared the length and genomic regional distribution of plasma cell-free DNA fragments in the healthy group and the stroke group and tried to identify the characteristic of cell-free DNA in the stroke patients, which could help to generate clinical values for assessing stroke disease status in the near future
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
