Use of synthetic oligonucleotides for determination of HTLV‐1 proviral load by real‐time PCR: a helpful alternative approach in the clinical management
Introduction of fluorescence-based Real-Time PCR (RT-PCR) also called qPCR is
increasingly used to detect multiple pathogens simultaneously and rapidly by
gene expression analysis of PCR amplification data. Real-time PCR data are
analyzed after setting an arbitrary threshold that must intersect the signal
curve in its exponential phase. The point at which the curve crosses the
threshold is called CT (Cycle Threshold). This simple and arbitrary value
however is not an elegant definition of CT value sometimes leads to
conclusions that are either false positive or negative. Therefore, the
purpose of this paper is to present a stable and consistent alternative
approach for the definition and determination of CT value that leads to near
zero false positives and negatives.
ABSTRACT
A quantitative real-time PCR assay was developed for the determination of antiviral drug susceptibility and growth kinetics of human herpesvirus 6. The susceptibility and fitness of a sensitive strain, HST, and its ganciclovir-resistant derivative, GCVR1, were then characterized, leading us to conclude that the mutations of this latter virus did not alter its fitness significantly.
Robust CT-prediction algorithm for RT-PCR