title: Single-channel multiplexing without melting curve analysis in real-time PCR.
abstract: Multiplex real-time PCR with quantification of targets in a single fluorescence channel has been the demand in biotechnology industry. Here, we develop a novel analytical real-time PCR technique to detect multiple targets in a single fluorescence channel without melting curve analysis. In this technique, we show the intensity of the fluorescence signals of two discrete Tm targets is different at certain temperatures called detection temperatures, by which a high Tm target can be detected regardless of a low Tm target. We then identify the low Tm target by utilizing a change of the fluorescence signals between two different detection temperatures. Furthermore, it enables us to determine quantification of each target in a single channel, possibly facilitating convenient patient care for drug treatment in clinics.
authors.affiliation: Seegene, Inc., Taewon Bldg., 91 Ogeum-ro, Songpa-Gu, Seoul 138-828, South Korea.
authors.forename: Young-Jo
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Seegene, Inc., Taewon Bldg., 91 Ogeum-ro, Songpa-Gu, Seoul 138-828, South Korea.
authors.forename: Daeyoung
authors.initials: D
authors.lastname: Kim
authors.affiliation: Seegene, Inc., Taewon Bldg., 91 Ogeum-ro, Songpa-Gu, Seoul 138-828, South Korea.
authors.forename: Kihoon
authors.initials: K
authors.lastname: Lee
authors.affiliation: Seegene, Inc., Taewon Bldg., 91 Ogeum-ro, Songpa-Gu, Seoul 138-828, South Korea.
authors.forename: Jong-Yoon
authors.initials: JY
authors.lastname: Chun
mesh_terms: D002690:Chlamydia Infections / Q000175:diagnosis / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D006069:Gonorrhea / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D025202:Molecular Diagnostic Techniques; D060885:Multiplex Polymerase Chain Reaction; D009344:Neisseria gonorrhoeae / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D044366:Transition Temperature
chemical_list: D004269:DNA, Bacterial