RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein.
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.
Ahn
Dae-Gyun
DG
Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
Jeon
Il-Ji
IJ
Kim
Jung Dong
JD
Song
Min-Sun
MS
Han
Seung-Ryul
SR
Lee
Seong-Wook
SW
Jung
Hyungil
H
Oh
Jong-Won
JW
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
D052157:Aptamers, Nucleotide; D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins