The present invention is directed to a method for performing a one-step RT-PCR for amplifying a target RNA comprising the steps (i) providing a sample which is supposed to contain said target RNA (ii) adding a reaction mixture comprising all reagents necessary to reverse transcribe said target RNA into cDNA and amplify at least a portion of said cDNA (iii) incubating said sample for a time interval of 0 seconds to 40 seconds at a temperature between 20°C and 65°C, and (iv) subjecting said sample to multiple cycles of a thermocycling protocol wherein the temperature of said sample is varied between at least a first temperature between 37°C and 72°C and a second temperature between 85°C and 100°C.
