Molecular diagnostic and predictive tests in the evolution of chronic hepatitis C anti-viral therapies

Abstract

Since the discovery of HCV, polymerase chain reaction (PCR) has significantly contributed to the understanding of the virus life cycle and its replicative kinetics during anti-viral therapy. Parallel to the progression of dual and triple combination treatment, real-time PCR molecular tests have constantly improved in their ability to monitor viral load and drive personalized management schedules. The current sensitivity, accuracy and dynamic range of the available assays fulfil the requirement of "companion diagnostics" and support the development of new directly acting antiviral (DAA)-based regimens.

Figure 1

Amperase® mediated selected inactivation of amplicons from previous reactions. Amplicons are synthetized during PCR using dUTP instead of dATP and made susceptible to cleavage by Amperase® which has no effect on sample, native DNA.

Figure 2

Progressive increase in sensitivity and dynamic ranges when proceeding from end point to real-time PCR. The latter allowed for a linear range increase of up to 4 logarithms.

Figure 3

Dual probe design of the Cobas® Ampliprep™/Cobas® Taqman™ HCV 2.0 test second version. The introduction of a second labelled probe and an additional reverse primers increased mismatchs tolerance and assay’s accuracy.