کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5031644 1470940 2017 14 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Digital quantification of gene methylation in stool DNA by emulsion-PCR coupled with hydrogel immobilized bead-array
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Digital quantification of gene methylation in stool DNA by emulsion-PCR coupled with hydrogel immobilized bead-array
چکیده انگلیسی
Aberrations of gene methylation in stool DNA (sDNA) is an effective biomarker for non-invasive colorectal cancer diagnosis. However, it is challenging to accurately quantitate the gene methylation levels in sDNA due to the low abundance and degradation of sDNA. In this study, a digital quantification strategy was proposed by combining emulsion PCR (emPCR) with hydrogel immobilized bead-array. The assay includes following steps: bisulfite conversion of sDNA, pre-amplification by PCR with specific primers containing 5′ universal sequences, emPCR of pre-amplicons with beaded primers to achieve single-molecular amplification and identification of hydrogel embedding beads coated with amplicons. The sensitivity and the specificity of the method are high enough to pick up 0.05% methylated targets from unmethylated DNA background. The successful detection of hypermethylated vimentin gene in clinical stool samples suggests that the proposed method should be a potential tool for non-invasive colorectal cancer screening.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 92, 15 June 2017, Pages 596-601
نویسندگان
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