In this kit, the extracted and purified nucleic acids are amplified directly using AP-RAD single-stage rRT-PCR ready-to-use reagents. In this process, the probe is connected to a specific target sequence located between the forward and reverse probes.
During the expansion phase of the PCR cycle, Taq polymerase 5′-nuclease activity degrades the probe, causing the reporter dye to separate from the quencher dye and generating a fluorescent signal. In each cycle, the extra molecules of the reporter dye are separated from the respective probe and increase the fluorescence intensity. The fluorescence intensity in each PCR cycle is recorded by the Real-PCR Instrument system and leads to the detection and announcement of the test result. This kit enables the detection of 3 genes N (nucleocapsid protein gene) and Replicase Open Reading Frame (ORF) 1b gene and internal control of RNase P with probe labels JOE, FAM, and CY5 (in some cases at the request of ROX diagnostic center), respectively.