COVID-19 (SARS-CoV-2) Triplex RT-qPCR Detection Kit

Research Use Only (RUO). This kit has not been tested and validated by any public health agency. This product is a multiplex fluorescent probe-based RT-qPCR assay system for the detection of COVID-19, the disease caused by the novel coronavirus SARS-CoV-2. 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 is 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, if during amplification of the template the probe is 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. Realtime monitoring of the entire PCR process can be assessed by monitoring the accumulation of fluorescent signals. This product provides triplex-detection in a single tube, including two independent genes of SARS-CoV-2 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 SARS-CoV-2's ORF1ab gene and N gene, respectively, avoiding nonspecific interference of SARS2003 and BatSARS-like virus strains. Internal control (RNAse P gene) provides a nucleic acid extraction procedural control and a secondary negative control. Positive control (SARS-CoV-2-pseudoviruse) provides a nucleic acid extraction and a reverse transcription control to validate the entire procedure and reagent integrity. Kit Contents/pack: Detection buffer 900µL × 2 tubes (Buffer, dNTPs, Primers, Probes). Enzyme mix 400µL × 1 tube (RNase Inhibitor, UDG, Reverse Transcriptase, Taq DNA polymerase). Positive Control 250µL × 1 tube (RNA pseudovirus containing target gene). Negative Control 250µL × 1 tube (DEPC-Treated Water). Instruments: Real-time PCR instrument with FAM, TEXAS RED/ROX and HEX/VIC channels, such as ABI7500, ABI Q3, ABI Q6, Roche LightCycler480, Bio-Rad CFX96