High-throughput ribozyme-based assays for detection of viral nucleic acids

Chemistry & Biology
Karl KossenScott D Seiwert

Abstract

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 x 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.

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Mar 24, 2009·Chemical Reviews·Juewen LiuYi Lu
Jul 24, 2004·Chemistry & Biology·John J Rossi
Jan 8, 2005·Chembiochem : a European Journal of Chemical Biology·Michael Famulok, Günter Mayer
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