2019-Novel Coronavirus (2019-nCoV) Triplex RT-qPCR Detection Kit
CATALOG NUMBER :CD302-02
100 tests / kit
This product is intended for the detection of 2019-Novel Coronavirus (2019-nCoV).
The detection result of this product is only for clinical reference, and it should not be used as the only evidence for clinical diagnosis and treatment.
PRINCIPLE OF DETECTION
This product is a multiplex fluorescent probe-based Taqman® RT-qPCR assay system. The Taqman fluorescent probe is a specific oligonucleotide based on a reporter-quencher mechanism. For each probe, the 5’-end is labeled with a fluorophore, while the 3’-end was labeled with a quencher.
When the probe is intact, the fluorescence emitted by the fluorophore is absorbed by the quencher, and no fluorescent signal is detected.
However, during amplification of the template, the probe will be degraded due to the 5′-3’ exonuclease activity of Taq DNA polymerase, and the fluorescent reporter and the quencher are cleaved and separated, then a fluorescent signal can be detected. The generation of each molecular
amplicon is accompanied by the generation of a fluorescent signal. Real time monitoring of the entire PCR process can be assessed by monitoring the accumulation of fluorescent signals.
This product provides triplex-detections in a single tube, including two independent genes of 2019-nCoV and an internal control which targets the human RNAse P (RNP) gene to assess specimen quality. Specific primers and probes were designed for the detection of conserved region of 2019-nCoV’s ORF1ab gene (RdRP region) and N gene, respectively, avoiding non-specific interference of SARS 2003 and Bat SARS-like virus strains. Internal control (RNAse P gene) provides a nucleic acid extraction procedural control and a secondary negative control. Positive control (2019-nCoV-pseudovirus) provides a nucleic acid extraction and a reverse transcription control to validate the entire procedure and reagent integrity.
|Detection Buffer||900 μL× 3 tubes||Buffer, dNTPs, Primers, Probes||Red|
|Enzyme Mix||400 μL× 1 tube||RNase Inhibitor, UDG, Reverse
Transcriptase, Taq DNA polymerase.
|Positive Control||250 μL× 1 tube||RNA pseudovirus containing target
|Negative Control||l 250 μL× 1 tube||DEPC-Treated Water||Green|
1. Do not mix the components from different batches for detection.
2. Additional Materials Required: Nucleic acid extraction reagents.
3. Nucleic acid extraction must be performed simultaneously with the Positive
control (2019-nCoV-pseudovirus) and Negative Control (DEPC-Treated Water) for monitoring the entire procedure to reduce false negative or false positive rates.
STORAGE & SHELF LIFE
All reagents should be stored at -30°C~-15°C with protection from light. The reagents are stable for 6 months when stored at the recommended condition.
The expiration date will not change if the kit is opened and stored at the recommended condition.
The expiration date will not change if the kit is transported with ice-packs for 4 days and/or treated with 10 freeze-thaw cycles.
Real-time PCR instrument with FAM, TEXAS RED/ROX and HEX/VIC channels, such as ABI 7500, ABI Q3, ABI Q6, Roche Lightcycler 480, Bio-Rad CFX96.