Issue 9, 2009

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 ofSARS-CoV N protein.

Graphical abstract: RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein

Article information

Article type
Paper
Submitted
06 Apr 2009
Accepted
13 Jul 2009
First published
21 Jul 2009

Analyst, 2009,134, 1896-1901

RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein

D. Ahn, I. Jeon, J. D. Kim, M. Song, S. Han, S. Lee, H. Jung and J. Oh, Analyst, 2009, 134, 1896 DOI: 10.1039/B906788D

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