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probes” can be classified into two major is totally efficient and all the reagents are
groups: hydrolysis and hybridization probes. available. The most important parameter in
Both types consist of an oligonucleotide, the RTi-PCR is the threshold cycle (CT value)
homologous to the internal region of the (Higuchi et al. 1992), which is used for the
amplicon, that is double-labeled, with a fluo- quantification of the sample. It corresponds
rophore or reporter dye (donor of fluores- to the cycle at which a statistically significant
cence) at the 5′-end and a quenching moiety increase in amplification-associated flores-
(acceptor of fluorescence) at the 3′-end. The cence is first detected, and is inversely cor-
distance between the fluorophores is a key related to the concentration of DNA present
factor in generating sequence-specific signals in the original sample (Walker 2002). The
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