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PubMed · 8643586

Extremely sensitive, background-free gene detection using binary probes and beta replicase.

S Tyagi·U Landegren·M Tazi·P M Lizardi·F R Kramer
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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 of target concentrations and it employs a universal format that can detect any infectious agent.

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Base SequenceCell LineDNA-Directed RNA PolymerasesGenes, ViralGenetic TechniquesHIV-1HumansLymphocytesMolecular Sequence DataNucleic Acid ConformationQ beta ReplicaseRNA ProbesRNA, MessengerRNA, ViralSensitivity and SpecificityTemplates, GeneticTranscription, GeneticJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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S Tyagi, U Landegren, M Tazi, P M Lizardi, F R Kramer. 1996-05-28. Extremely sensitive, background-free gene detection using binary probes and beta replicase.. https://doi.org/10.1073/pnas.93.11.5395

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