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Extremely sensitive, background-free gene detection using binary probes and Qf8 replicase (diagnostic clinical assays/isolation of probe-target hybrids/T4 DNA ligase/target-dependent signal generation/exponential amplification of reporter RNAs)
Journal article

Extremely sensitive, background-free gene detection using binary probes and Qf8 replicase (diagnostic clinical assays/isolation of probe-target hybrids/T4 DNA ligase/target-dependent signal generation/exponential amplification of reporter RNAs)

Sanjay Tyagi, Ulf Landegren, Mounssef Tazi, Paul M. Lizardi and Fred Russell Kramer
Proceedings of the National Academy of Sciences (PNAS), Vol.93, pp.5395-5400
05/1996

Abstract

We have developed a specific and sensitive nucleic acid amplification assay that is suitable for routine gene detection. The assay is based on a novel molecular genetic strategy in which two different RNA probes are hybridized to adjacent positions on a target nucleic acid and then ligated to form an amplifiable reporter RNA. The reporter RNA is then replicated up to a hundred billion-fold in a 30-min isothermal reaction that signals the presence of the target. The assay can detect fewer than 100 nucleic acid molecules; it provides quantitative results over a wide range oftarget concentrations and it employs a universal format that can detect any infectious agent.
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https://www.pnas.org/doi/pdf/10.1073/pnas.93.11.5395View
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