title: Label-free detection of bacterial RNA using polydiacetylene-based biochip.
abstract: We developed a simple and effective polydiacetylene-based, label-free multiplex DNA chip for the detection of various pathogenic microorganisms. A novel immobilization method of PDA vesicles on glass slides was exploited using α-cyclodextrin (α-CD). The surface topography of the efficiently immobilized PDA vesicles was confirmed using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Then, oligonucleotides complementary to rRNAs of three pathogenic bacteria were conjugated to the PDA vesicles. Finally, crude lysate of pathogenic bacteria was applied to the PDA biochip. The pathogenic bacteria were specifically detected by DNA-RNA hybridization in an hour. The new PDA sensor was effective in detecting multiple pathogenic bacteria easily and accurately without rigorous purification, amplification, and labeling of their genetic components.
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Moo-Kyung
authors.initials: MK
authors.lastname: Park
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Kyung-Woo
authors.initials: KW
authors.lastname: Kim
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul 136-713, Republic of Korea. Electronic address: ahn@korea.ac.kr.
authors.forename: Dong June
authors.initials: DJ
authors.lastname: Ahn
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul 136-713, Republic of Korea. Electronic address: mkoh@korea.ac.kr.
authors.forename: Min-Kyu
authors.initials: MK
authors.lastname: Oh
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods; D015342:DNA Probes / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D000078789:Polyacetylene Polymer; D011108:Polymers; D053279:Polyynes; D012329:RNA, Bacterial / Q000032:analysis* / Q000235:genetics; D012477:Salmonella enteritidis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D015342:DNA Probes; D000078789:Polyacetylene Polymer; D011108:Polymers; D012329:RNA, Bacterial; D053279:Polyynes; C082361:polydiacetylene