The four steps below give an overview of the technology behind MeltPlex®. If you would like more information please contact us.
PCR primers for amplification of DNA or reverse transcribed RNA target serve as target DNA. An amplification step is performed using PCR primers and multiple target-specific MeltPlex® probes labeled with fluorescent dyes.
If the specific target is present, the 5’ quencher is cleaved from the probe which is now activated (fluorescent).
A second, common, quenching probe hybridizes to the Melting Tag region and generate a melting curve with a profile specific for each target-specific probe.
The detection of the activated probes is done by melting curve analysis on a Real-Time PCR system. 20+ target-specific MeltPlex® detection probes, labeled by multiple fluorophores and varying in Melting Tag temperature, enable specific detection of amplified product and the corresponding pathogen.
16 MeltPlex® probes were converted from exisiting TaqMan probe designs1 to provide a 16-plex probe mix for detection of DNA of 17 different Hemorrhagic fever viruses.
Pang, Zheng et al. "Comprehensive Multiplex One-Step Real-Time TaqMan qRT-PCR Assays for Detection and Quantiﬁcation of Hemorrhagic Fever Viruses." Ed. Stefan Dﬁbel. PLoS ONE 9.4 (2014): e95635. PMC. Web. 2 Apr, 2017.