High-Throughput Ribozyme-Based Assays for Detection of Viral Nucleic Acids
| Autor: | Narendra K. Vaish, Christopher Pasko, Karl Kossen, Vasant Jadhav, Hong Wang, Barry Polisky, Robert D. Jenison, Scott D Seiwert, James McSwiggen |
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| Rok vydání: | 2004 |
| Předmět: |
Hepatitis C virus
Molecular Sequence Data Clinical Biochemistry Enzyme-Linked Immunosorbent Assay Biology medicine.disease_cause Sensitivity and Specificity Biochemistry Drug Discovery medicine Oligoribonucleotides RNA Catalytic Molecular Biology Immunoassay Pharmacology Detection limit chemistry.chemical_classification DNA ligase Base Sequence medicine.diagnostic_test Ribozyme RNA General Medicine Hepatitis C Kinetics chemistry Nucleic acid biology.protein Nucleic Acid Conformation RNA Viral Molecular Medicine |
| Zdroj: | Chemistry & Biology. 11:807-815 |
| ISSN: | 1074-5521 |
| Popis: | Many reports have suggested that target-activated ribozymes hold potential value as detection reagents. We show that a "half"-ribozyme ligase is activated similarly by three unstructured oligoribonucleotides representing the major sequence variants of a hepatitis C virus 5′-untranslated region (5′-UTR) target and by a structured RNA corresponding to the entire 5′-UTR. Half-ribozyme ligation product was detected both in an ELISA-like assay and in an optical immunoassay through the use of hapten-carrying substrate RNAs. Both assay formats afford a limit of detection of approximately 1 × 10 6 HCV molecules (1.6 attomol, 330 fM), a sensitivity which compares favorably to that provided by standard immunoassays. These data suggest that target-activated ribozyme systems are a viable approach for the sensitive detection of viral nucleic acids using high-throughput platforms. |
| Databáze: | OpenAIRE |
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