Jump to main content
Jump to site search
PLANNED MAINTENANCE Close the message box

Scheduled maintenance work on Wednesday 22nd May 2019 from 11:00 AM to 1:00 PM (GMT).

During this time our website performance may be temporarily affected. We apologise for any inconvenience this might cause and thank you for your patience.

Issue 13, 2018
Previous Article Next Article

In vitro diagnosis of DNA methylation biomarkers with digital PCR in breast tumors

Author affiliations


Liquid biopsy of cancers using DNA methylation biomarkers has received significant interest, where the quantification of multiple biomarkers is generally needed for improving the sensitivity and specificity of cancer diagnosis. However, the inefficiency of the traditional quantitative polymerase chain reaction (qPCR)-based MethyLight assay for detecting the extremely low concentration of methylated DNA fragments in body fluids limits its clinical applications. Here, we developed an ultrasensitive microwell chip digital polymerase chain reaction (dPCR)-based MethyLight assay. Using the synthesized samples, the developed MethyLight assay can achieve 103–104-fold lower limit of detection and 1–16-fold lower limit of quantification than the traditional MethyLight assay. Four hypermethylated alleles (RARβ2, BRCA1, GSTP1 and RASSF1A) related to breast cancer in twenty-three clinical samples were tested using the microwell chip dPCR-based MethyLight assay. The results showed that the dPCR assay achieves ∼2 times enhancement in the cancer detection rate over the traditional quantitative PCR. Furthermore, the dPCR can detect the healthy and benign samples, which are undetectable using the traditional MethyLight assay. In multiple gene analysis, we achieved the highest detection rate of 93.3% (in the “OR” format of RARβ2 and GSTP1). Lastly, the estimated cut-off values in the dPCR assay were: <1, ∼1 to 100 and >100 (copies per μL) referring to the healthy, benign and malignant breast cancers, respectively. Therefore, the developed microwell chip dPCR-based MethyLight assay could provide a powerful tool for cancer biopsy diagnosis and disease monitoring.

Graphical abstract: In vitro diagnosis of DNA methylation biomarkers with digital PCR in breast tumors

Back to tab navigation

Supplementary files

Publication details

The article was received on 01 Feb 2018, accepted on 15 Apr 2018 and first published on 18 Apr 2018

Article type: Paper
DOI: 10.1039/C8AN00205C
Analyst, 2018,143, 3011-3020

  •   Request permissions

    In vitro diagnosis of DNA methylation biomarkers with digital PCR in breast tumors

    X. Cui, L. Cao, Y. Huang, D. Bai, S. Huang, M. Lin, Q. Yang, T. J. Lu, F. Xu and F. Li, Analyst, 2018, 143, 3011
    DOI: 10.1039/C8AN00205C

Search articles by author