The Coronavirus COVID-19 (CE IVD) genesig® Real-Time PCR assay is an in vitro diagnostic test based on Real-Time PCR technology, developed for specific detection of SARS-CoV-2 viral RNA. The probe system is based on the standard hydrolysis probe system known as TaqMan® Technology. The COVID-19 specific probe is labelled with the FAM fluorophore and the internal control is labelled with the HEX fluorophore.
The assay includes an internal control to identify possible PCR inhibition, measure extraction purity and confirm the integrity of the PCR run.
Real-Time PCR technology utilizes polymerase chain reaction (PCR) for the amplification of specific target sequences and target specific probes for the detection of the amplified RNA. The probes are labelled with fluorescent reporter and quencher dyes.
The COVID-19 Positive control template (PCT) tube contains a standard number of copies of SARSCoV-2 RNA (Coronavirus COVID-19) specific sequence. Following resuspension, this will give a concentration 1.67 x 105 copies per µl. To ensure PCR run validity, the PCT should produce Cq value 14-22 in the FAM channel.
Extraction Kits / Instruments
The Coronavirus COVID-19 (CE IVD) genesig® Real-Time PCR kit was developed to be used with an extraction system validated as a CE IVD device for use in the extraction of RNA (either exclusively, or with DNA) from clinical samples, including nasopharyngeal swabs, oropharyngeal swabs and sputum. Validation of the assay was performed on the automated extraction system GenoXtract® from HAIN Lifescience GmbH (Brucker) using the GXT DNA/RNA Extraction kit (CE IVD).
Real-Time PCR instruments
The genesig® Real-Time PCR Coronavirus COVID-19 (CE IVD) was developed and validated to be used with the following Real-Time PCR instruments.
• Applied Biosystem® 7500 Real-Time PCR System (software version 2.3)
• Roche® LightCycler 480 II (software version 1.5)
• Bio-Rad CFX Connect™Real-Time PCR Detection System (Maestro software version 1.1)