title: Upconversion Nanoparticle-Based Förster Resonance Energy Transfer for Detecting DNA Methylation.
abstract: Aberrant methylation of a crucial CpG island is the main mechanism for the inactivation of CDKN2A in the early stages of carcinogenesis. Therefore, the detection of DNA methylation with high sensitivity and specificity is important, and various detection methods have been developed. Recently, upconversion nanoparticles (UCNPs) have been found to display a high signal-to-noise ratio and no photobleaching, making them useful for diagnostic applications. In this pilot study, we applied UCNPs to the detection of CDKN2A methylation and evaluated the feasibility of this system for use in molecular diagnostics. DNA PCR was performed using biotinylated primers, and the PCR amplicon was then intercalated with SYTOX Orange dye, followed by incubation with streptavidin-conjugated UCNPs. Fluorescence detection of the Förster resonance energy transfer (FRET) of the UCNPs (MS-UC-FRET) was then performed, and the results were compared to those from real-time PCR (RQ-PCR) and pyrosequencing. Detection by MS-UC-FRET was more sensitive than that by either RQ-PCR or pyrosequencing. Our results confirmed the success of our MS-UC-FRET system for detecting DNA methylation and demonstrated the potential application of this system in molecular diagnostics.
authors: Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul 05505, Korea. sj1kim1@gmail.com.
authors: Seockjune
authors: S
authors: Kim
authors: Department of Laboratory Medicine, Center for Diagnostic Oncology, Research Institute and Hospital, National Cancer Center, Goyang-si 10408, Korea. mindcatch@ncc.re.kr.|Hematologic Malignancy Branch, Research Institute and Hospital, National Cancer Center, Goyang-si 10408, Korea. mindcatch@ncc.re.kr.
authors: Sang-Hyun
authors: SH
authors: Hwang
authors: Department of Laboratory Medicine, Center for Diagnostic Oncology, Research Institute and Hospital, National Cancer Center, Goyang-si 10408, Korea. relationship01@ncc.re.kr.
authors: Su-Gyeong
authors: SG
authors: Im
authors: Department of Internal Medicine, Pusan National University School of Medicine and Biomedical Research Institute, Pusan National University Hospital, 179 Gudeok-ro, Seo-gu, Busan 602-739, Korea. leemk@pusan.ac.kr.
authors: Min-Ki
authors: MK
authors: Lee
authors: Department of Pathology, Pusan National University School of Medicine and Biomedical Research Institute, Pusan National University Hospital, 179 Gudeok-ro, Seo-gu, Busan 602-739, Korea. cnlee@pusan.ac.kr.
authors: Chang-Hun
authors: CH
authors: Lee
authors: Department of Chemistry, Gachon University, Seongnam, Gyeonggi, and Gachon Medical Research Institute, Gil Medical Center, Inchon 461-701, Korea. triaza@gmail.com.
authors: Sang Jun
authors: SJ
authors: Son
authors: Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul 05505, Korea. hboh@amc.seoul.kr.
authors: Heung-Bum
authors: HB
authors: Oh
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D063646:Carcinogenesis / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050763:Cyclin-Dependent Kinase Inhibitor p18 / Q000235:genetics / Q000302:isolation & purification*; D019175:DNA Methylation / Q000235:genetics*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D006046:Gold / Q000737:chemistry; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry; D057089:Pathology, Molecular / Q000379:methods*; D059629:Signal-To-Noise Ratio
chemical_list: C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050763:Cyclin-Dependent Kinase Inhibitor p18; D006046:Gold
keywords: CDKN2A; DNA methylation; FRET; MS-UC-FRET; MSP; biosensing; pyrosequencing; upconversion nanoparticle