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Isothermal amplification is a rapid, simple alternative to PCR, with amplification commonly detected using fluorescently labelled oligonucleotide probes, intercalating dyes or increased turbidity as a result of magnesium pyrophosphate generation. SNP identification is possible but requires either allele-specific primers or multiple dye-labelled probes, but further downstream processing is often required for allelic identification. Here we demonstrate that modification of common isothermal amplification methods by the addition of HyBeacon probes permits homogeneous sequence detection and discrimination by melting or annealing curve analysis. Furthermore, we demonstrate that isothermal amplification and sequence discrimination is possible directly from a crude sample such as an expressed buccal swab.

Original publication

DOI

10.1016/j.mcp.2014.12.001

Type

Journal article

Journal

Mol Cell Probes

Publication Date

04/2015

Volume

29

Pages

92 - 98

Keywords

CPA, HyBeacons, Isothermal amplification, LAMP, Probes, SMAP, Chlamydia trachomatis, Humans, Molecular Probes, Nucleic Acid Amplification Techniques, Polymorphism, Single Nucleotide, Sequence Analysis, DNA, Vitamin K Epoxide Reductases