title: A simple colorimetric detection of DNA methylation.
abstract: In this work, we describe a simple colorimetric method to detect DNA methylation. Adenomatous polyposis coli (APC) with a small CpG region containing methylated cytosine (methylated APC) was synthesized and tested. Methylated APC was first captured and enriched by anti-5-methylcytosine monoclonal antibody conjugated magnetic microspheres (MMPs). Then a probe partly complementary to the APC sequence was added, resulting in the formation of DNA duplexes. The microsphere-captured probe was then released by heat denaturation and added into unmodified gold nanoparticle (AuNP) solution. Colorimetric detection was performed by salt-induced aggregation. The limit of detection is 80 fmol. Semi-quantitative analysis was done with a UV/Vis spectrophotometer by recording the absorbance of AuNP solution at 520 nm. Thus, this method provides a simple, rapid and quantitative tool for DNA methylation detection.
authors.affiliation: University of Science and Technology of China, Hefei, 230026, China.
authors.forename: Chenchen
authors.initials: C
authors.lastname: Ge
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Fang
authors.forename: Junhua
authors.initials: J
authors.lastname: Chen
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.forename: Xuewen
authors.initials: X
authors.lastname: Lu
authors.forename: Lingwen
authors.initials: L
authors.lastname: Zeng
mesh_terms: D025601:Adenomatous Polyposis Coli Protein / Q000235:genetics / Q000378:metabolism; D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry / Q000379:methods*; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D005240:Feasibility Studies; D006046:Gold / Q000737:chemistry; D059346:Magnets / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D008863:Microspheres
chemical_list: C555337:APC protein, human; D025601:Adenomatous Polyposis Coli Protein; D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D006046:Gold