title: Upconversion nanoparticle-based Förster resonance energy transfer for detecting the IS6110 sequence of Mycobacterium tuberculosis complex in sputum.
abstract: Upconversion nanoparticles (UCNPs), which are excited at near-infrared wavelength (980 nm), emit high-energy photons. Since UCNPs display a high signal-to-noise ratio and no photobleaching, they are extremely useful for diagnostic application. In this study, we applied UCNPs for detecting the IS6110 sequence of the Mycobacterium tuberculosis complex (MTBC) and evaluated the feasibility of the system for use in molecular diagnostics. Using biotinylated primers, IS6110 DNA PCR was performed and the PCR amplicon was then mixed with streptavidin-conjugated UCNPs, followed by intercalation with SYTOX Orange dye. Fluorescence detection for the Förster resonance energy transfer (FRET) of the UCNPs (UCNP-FRET) was then performed. The estimated lowest detection by UCNP-FRET was 10(2) copies/μL of IS6110 DNA (157 bp). The kappa agreement of the UCNP-FRET assay with conventional PCR was 0.8464 (95% confidence interval, 0.7442-0.9486) and false-negative results were reduced. Our results demonstrated the successful implementation of the UCNP-FRET system in detecting the IS6110 sequence of the MTBC and its potential application for molecular diagnostics.
authors.affiliation: Department of Laboratory Medicine, Center for Diagnostic Oncology, Research Institute and Hospital, National Cancer Center, Goyang-si 410-769, Republic of Korea; Hematologic Malignancy Branch, Research Institute, National Cancer Center, Republic of Korea.
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Hwang
authors.forename: Su-Gyeong
authors.initials: SG
authors.lastname: Im
authors.forename: Heungsup
authors.initials: H
authors.lastname: Sung
authors.forename: Sang Soo
authors.initials: SS
authors.lastname: Hah
authors.forename: Vu Thanh
authors.initials: VT
authors.lastname: Cong
authors.forename: Do-Hoon
authors.initials: DH
authors.lastname: Lee
authors.forename: Sang Jun
authors.initials: SJ
authors.lastname: Son
authors.forename: Heung-Bum
authors.initials: HB
authors.lastname: Oh
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000302:isolation & purification; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000302:isolation & purification*; D053758:Nanoparticles; D013183:Sputum / Q000382:microbiology*; D014376:Tuberculosis / Q000175:diagnosis*
chemical_list: D004269:DNA, Bacterial
keywords: Förster resonance energy transfer; IS6110 PCR; Mycobacterium tuberculosis complex; Upconversion; Upconversion nanoparticles