A METHOD FOR SUPPRESSING NON-SPECIFIC AMPLIFICATION PRODUCTS IN NUCLEIC ACID AMPLIFICATION TECHNOLOGIES

摘要

The use of Nucleic Acid Amplification Technologies (NAATs) to rapidly copy a specific fragment of DNA from a few starting molecules is used to detect that DNA in a sample, Applications include identification of a pathogen in a clinical sample. Non-specific DNA amplification occurring in the absence of any input DNA template is frequently observed after many amplification cycles or in the case of isothermal amplification, with time. The disclosed embodiments describe the surprising finding that the amplification of primer-only artefacts is suppressed in reactions where a fraction of the oligos involved in the reaction contain 3' blocked terminal bases that cannot support extension by DNA polymerases. The effect of these 3' blocked oligos is to disproportionality retard the primer-only reactions compared to targeted templated primed true positive reactions, thus opening up the window separating false positives from true positives and vastly improving the reaction specificity and sensitivity.