title: RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein.
abstract: Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of a newly emerged disease SARS. The SARS-CoV nucleocapsid (N) protein is one of the most abundant structural proteins and serves as a diagnostic marker for accurate and sensitive detection of the virus. Using a SELEX (systematic evolution of ligand by exponential enrichment) procedure and recombinant N protein, we selected a high-affinity RNA aptamer capable of binding to N protein with a dissociation constant of 1.65 nM. Electrophoretic mobility shift assays and RNA competition experiments showed that the selected aptamer recognized selectively the C-terminal region of N protein with high specificity. Using a chemiluminescence immunosorbent assay and a nanoarray aptamer chip with the selected aptamer as an antigen-capturing agent, we could sensitively detect N protein at a concentration as low as 2 pg/ml. These aptamer-antibody hybrid immunoassays may be useful for rapid, sensitive detection of SARS-CoV N protein.
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Dae-Gyun
authors.initials: DG
authors.lastname: Ahn
authors.forename: Il-Ji
authors.initials: IJ
authors.lastname: Jeon
authors.forename: Jung Dong
authors.initials: JD
authors.lastname: Kim
authors.forename: Min-Sun
authors.initials: MS
authors.lastname: Song
authors.forename: Seung-Ryul
authors.initials: SR
authors.lastname: Han
authors.forename: Seong-Wook
authors.initials: SW
authors.lastname: Lee
authors.forename: Hyungil
authors.initials: H
authors.lastname: Jung
authors.forename: Jong-Won
authors.initials: JW
authors.lastname: Oh
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide; D002522:Chlorocebus aethiops; D000086462:Coronavirus Nucleocapsid Proteins; D007118:Immunoassay / Q000379:methods*; D008163:Luminescent Measurements; D046228:Microarray Analysis; D019590:Nucleocapsid Proteins / Q000032:analysis*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D014709:Vero Cells
chemical_list: D052157:Aptamers, Nucleotide; D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins