Abstract: The invention is a method to determine the amounts, in particular the relative amounts, of nucleic acids in complex biological samples by means of real-time PCR. According to the invention the biological sample is systematically diluted and each dilution is studied by real-time PCR for all genes of interest. From the dependence of the threshold cycle on dilution factor for each of the genes, the PCR efficiencies of the reactions are determined in the particular samples. Determining also the relative sensitivity of the real-time PCR assays compared, the relative amounts of two nucleic acids in complex biological samples are determined with unprecedented accuracy.

Abstract: The invention is a method to determine the amounts, in particular the relative amounts, of nucleic acids in complex biological samples by means of real-time PCT. According to the invention the biological sample is systematically diluted and each dilution is studied by real-time PCR for all genes of interest. From the dependence of the threshold cycle on dilution factor for each of the genes, the PCR efficiencies of the reactions are determined in the particular samples, determining also the relative sensitivity of the real-time PCR assays compared, the relative amounts of two nucleic acids in complex biological samples are determined with unprecedented accuracy.