I have a question about the LAMP reaction regarding the need to add Reverse Transcriptase enzyme for virus (RNA) detection. Is the addition of the RT step crucial in the LAMP method from direct tissue?
Let's say if we need to diagnose a presence of virus infection from a tissue sample that the genetic material (RNA) is extracted using the boiling lysis method (95C for 5 min, then directly used as template in LAMP reaction) is the cDNA synthesis using RTase is crucial for the diagnostic process? or it will be ok just to use the direct method (without RT step and just using FiP/BiP, F3/B3, and LF/LB primer without d(T)). Is it will significantly affect the sensitivity of the test?