Absolute quantification of DNA methylation using microfluidic chip-based digital PCR

Biosensors & Bioelectronics
Zhenhua WuJianlong Zhao

Abstract

Hypermethylation of CpG islands in the promoter region of many tumor suppressor genes downregulates their expression and in a result promotes tumorigenesis. Therefore, detection of DNA methylation status is a convenient diagnostic tool for cancer detection. Here, we reported a novel method for the integrative detection of methylation by the microfluidic chip-based digital PCR. This method relies on methylation-sensitive restriction enzyme HpaII, which cleaves the unmethylated DNA strands while keeping the methylated ones intact. After HpaII treatment, the DNA methylation level is determined quantitatively by the microfluidic chip-based digital PCR with the lower limit of detection equal to 0.52%. To validate the applicability of this method, promoter methylation of two tumor suppressor genes (PCDHGB6 and HOXA9) was tested in 10 samples of early stage lung adenocarcinoma and their adjacent non-tumorous tissues. The consistency was observed in the analysis of these samples using our method and a conventional bisulfite pyrosequencing. Combining high sensitivity and low cost, the microfluidic chip-based digital PCR method might provide a promising alternative for the detection of DNA methylation and early diagnosis of epigenetics-r...Continue Reading

Citations

Feb 2, 2018·Critical Reviews in Clinical Laboratory Sciences·Amogha TadimetyJohn X J Zhang
Jul 25, 2019·Lab on a Chip·Chengyu DengChang Lu
Dec 24, 2019·Critical Reviews in Clinical Laboratory Sciences·Connie WuDavid R Walt
Dec 19, 2020·Biomolecules·Francesca GalardiMatteo Benelli
Oct 20, 2018·ACS Applied Materials & Interfaces·Junjie ChiZhongze Gu
Dec 4, 2021·Scientific Reports·Abbas JaliliAmir Hossein Kazemipour Ashkezari

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