Absolute quantification of specific nucleic acids by RT-PCR using a nonlinear mathematical model for data analysis

Quantification of specific nucleic acids plays a key role in the study of gene expression and in the clinical analysis of viral loads such as for human immunodeficiency virus (HIV-1) and hepatitis C virus (HCV). Among methods developed for this purpose, competitive (RT)-PCR, ¹ [polymerase chain reaction (PCR), with or without prior reverse transcription (RT)] is the most popular for its high sensitivity and relatively low cost. However, in the past decade the method has not improved much to overcome its main drawbacks: the difficulty of generating valid internal standards ^2-4 and the controversy over its accuracy. ^4-12.