SARS-COV-2 diagnosis by phi29 polymerase amplification

SARS-COV-2 diagnosis by phi29 polymerase amplification The project aims the validation of a reliable method for diagnosis of SARS-COV-2 infection. This novel method will allow the improved detection of the viral RNA molecule by means of a virus-specific linear probe that becomes circularized only in the presence of the viral RNA. Once the probe becomes circular, an engineered form of phi29 DNA polymerase (CSIC patent licensed to 4basebio) will start copying the circular template by generating an unlimited synthesis reaction defined as "rolling-circle replication" (RCR), which depends on the unique and robust strand-displacement synthesis capacity of this phi29 DNA polymerase variant. This peculiar mode of synthesis, a hallmark of the isothermal DNA amplification of full genomes used worldwide, could be easily detected as a readout of the presence of SARS-COV-2 genomic RNA