Norovirus RT-PCR Kit

The Norovirus RT-PCR Kit has been tested on the Roche LightCycler 480II, Agilent Technologies Mx3005P, Applied Biosystems ABI 7500, Bio-Rad CFX96, and QIAGEN Rotor-Gene Q real-time PCR instruments. Norovirus IV will also be detected by the Norovirus RT-PCR Kit. Contact ALPCO for additional real-time PCR instrument information.  The norovirus RT-PCR is a multiplex real-time PCR assay for the direct, qualitative detection of norovirus (genogroup GI and GII) in human stool samples. Detection takes place in one-step realtime RT-PCR format, where reverse transcription (RT) and the subsequent PCR takes place in the same reaction tube. The isolated RNA is transcribed into cDNA using a reverse transcriptase. Subsequently amplification of the gene fragments specific for norovirus GI and GII occurs if present. The amplified targets (ORF1/ORF2 junction region) are detected with hydrolysis probes, which are labeled at one end with a quencher and at the other end with a fluorescent reporter dye (fluorophore). In the presence of a target, the probes hybridize to the amplicons. During the extension step, the Taq-Polymerase breaks the reporter-quencher proximity. The reporter emits a fluorescence signal which is detected by the optical unit of a real-time PCR instrument. The fluorescence signal increases with the amount of formed amplicons. The norovirus RT-PCR assay contains an Internal Control RNA (ICR) as an internal control of the sample preparation procedure and to determine possible PCR inhibition.