Improved isothermal amplification

Abstract

The invention relates to a method of reducing the incidence of false positives, increasing efficiency, and/or increasing the sensitivity of loop-mediated isothermal amplification (LAMP) of a target nucleic acid comprising combining: a plurality of LAMP primers for a target nucleic acid, wherein the plurality of LAMP primers comprises a F3 primer and a B3 primer, wherein both the F3 and B3 primer or the F3 primer comprises a promoter for an RNA polymerase at the 5′ end; dNTPs; a strand-displacing DNA polymerase; a reverse transcriptase; an RNA polymerase; and rNTPs; and optionally a nucleic acid binding dye or probe, to form a LAMP reagent mix, incubating the LAMP reagent mix with the target nucleic acid under conditions suitable for amplification of the target nucleic acid with increased efficiency and/or sensitivity compared to the same amplification performed in the absence of the promoter, RNA polymerase and rNTPs.

Dependencies are inferred from the claim text.