LIONPROBESTM : ”A new fluorescent probe system for real-time detection, quantification, and nucleic acid sequence analysis” (Patent PCT ES2005/070093)

LIONPROBESTM represent an innovative fluorescent system for detecting, quantifying, and analysis specific nucleic acid sequences. The amplification-detection system depends on a non- conventional DNA polymerase enzyme, a custom-probe design, and technology for high resolution melting.

In contrast to hydrolysis probes that rely on the 5’-3’ exonuclease activity of conventional DNA polymerases, LIONPROBESTM exploit the 3’-5’ exonuclease activity present in a proofreading polymerase. In this way, a dually marched oligonucleotide probe, containing a fluorescent marker and quenching molecule, undergoes cleavage and initiates the extension of the leading strand through incorporation of dNTP units. Interesting, the cleavage and subsequent DNA extension only occurs when the market oligonucleotide presents some sequence dissimilarity in the 3’ end to the complimentary template strand (Figure 1).

A further application of the LIONPROBESTM system is that when fluorescent market is incorporated in the amplification product, the subsequent product undergoes a release of fluorescence energy signal when the product changes from double stranded product to single stranded product. Using high-resolution melting software of appropriate real-time DNA analyser, the change of fluorescence is detected as a “reverse melting peak” where fluorescence increases at the specific temperature where strand dissociation of the amplification product occurs.

LionprobesTM are innovators fluorescent probes for detecting, quantifying, and analysis specific nucleic acid by real time PCR. Our company has been developed an important number of products based on LionprobesTM technology.

Figure 1: LionprobesTM technology