The virus SARS-CoV-2 (previously 2019-nCoV) is of great interest as it is responsible for the coronavirus disease COVID-19. Consequently, the development of corresponding products is currently running at full speed. Hereby we announce the launch of Assay Genie´s COVID-19 (SARS-CoV-2) Triplex RT-qPCR Detection Kit.
COVID-19 (SARS-CoV-2) Triplex RT-qPCR Detection Kit
This product is a multiplex fluorescent probe-based RT-qPCR assay system for the detection of COVID-19. 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.
| Component | Amount | Ingredients |
| Detection Buffer | 900 μl × 2 tubes | Buffer, dNTPs, Primers, Probes |
| Enzyme Mix | 400 µl × 1 tube | RNase Inhibitor, UDG, Reverse Transcriptase, Taq DNA Polymerase |
| Positive Control | 250 µl × 1 tube | RNA pseudovirus containing target gene |
| Negative Control | 250 µl × 1 tube | DEPC-Treated Water |
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.
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Detection limitation: 200 copies/ml
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Precision: using precision reference CV1 and CV2 for within-batch and between-batch detection, the coefficient of variation (CV) of their Ct values is ≤5.0%
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Conformity rate of Negative Control: 100%
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Conformity rate of Positive Control: 100%
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Specificity: non-specific interference of Influenza A Virus (H1N1, H3N2,H7N9, H5N1), Influenza B Virus (Yamagata, Victoria), Respiratory Syncytial Virus (type B), Respiratory Adenovirus (type 3, type 7), Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, etc.
