Principle of Inspection This product is a multiplex fluorescent probe-based Taqman RT-qPCR assay system. Specific primers and probes were designed for the detection of conserved region of virus ORF1ab gene and N gene, respectively. An Internal control serves as the monitor to confirm successful extraction and identify possible PCR inhibition for the entire testing system to prevent false negative detection results.
Specimen Processing (Specimen Processing area)

2.1 Nucleic acid extraction
It is recommended to take 200 μL liquid samples, Positive Control and Negative Control fo rBioteke Corporation
nucleic acid extraction, according to the corresponding requirements and procedures of viral
RNA extraction kits.
Note: Nucleic acid was extracted by adding 2 μL internal control to the samples, Positive
Control and Negative Control.

2.2 Template Addition
Add 5 μL of extracted Positive Control products , 5 μL of extracted Negative Control
products, and 5 μL of extracted RNA from specimen to different PCR reaction tubes which
contained 20 μL of PCR mix. The total volume is 25 μL. Cap the reaction tubes tightly,
centrifuge them at low speed. Then, move to the Detection Area.

3.
PCR Amplification (Detection Area)
3.1 Put the reaction tubes on a PCR instrument and label the sample name, Positive Control
and Negative Control in the corresponding sequence.
3.2 Settings of detection fluorescence: (1) ORF1ab gene (FAM);(2) N gene((HEX);
(3)Internal Control (ROX).