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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2009 |
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| Erschienen: |
2009 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:134 |
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| Enthalten in: |
The Analyst - 134(2009), 9 vom: 18. Sept., Seite 1896-901 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Ahn, Dae-Gyun [VerfasserIn] |
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| Links: |
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| Themen: |
Aptamers, Nucleotide |
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| Anmerkungen: |
Date Completed 04.03.2010 Date Revised 07.12.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1039/b906788d |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM190713410 |
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| 500 | |a Date Revised 07.12.2022 | ||
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| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Aptamers, Nucleotide |2 NLM | |
| 650 | 7 | |a Coronavirus Nucleocapsid Proteins |2 NLM | |
| 650 | 7 | |a Nucleocapsid Proteins |2 NLM | |
| 700 | 1 | |a Jeon, Il-Ji |e verfasserin |4 aut | |
| 700 | 1 | |a Kim, Jung Dong |e verfasserin |4 aut | |
| 700 | 1 | |a Song, Min-Sun |e verfasserin |4 aut | |
| 700 | 1 | |a Han, Seung-Ryul |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Seong-Wook |e verfasserin |4 aut | |
| 700 | 1 | |a Jung, Hyungil |e verfasserin |4 aut | |
| 700 | 1 | |a Oh, Jong-Won |e verfasserin |4 aut | |
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