The POLGEN SARS-CoV-2 (COVID-19) Multiplex qPCR Detection Kit is an in vitro diagnostic test, based on real-time polymerase chain reaction technology. It tests for the presence or absence of ribonucleic acid (RNA) of SARS-CoV-2 coronavirus, specifically, in lower and upper respiratory tract samples from patients suspected of COVID-19 viral infection.

POLGEN SARS-CoV-2 (COVID-19) Multiplex qPCR Detection test includes a Primer-Probe set inside the Reaction Mix that contains primers and probes for human RNaseP and viral N1 and N2 regions. The internal control (primer/probe set for human RNaseP gene) is used to identify possible qPCR inhibition, confirm the integrity of the reagents, and verify the quality of RNA extraction from patient material. RNaseP primers and probes included in the kit recognize the human RNaseP gene. The primers and probes for viral N1 and N2 regions specifically identify two distinct regions on the nucleocapsid (N) gene of the SARS-CoV-2 virus. These primers and probes do not recognize the SARS-CoV virus, whose genetic structure is highly similar to SARS-CoV-2.

POLGEN SARS-CoV-2 (COVID-19) Multiplex qPCR Detection test also includes a positive control, which is labelled as “Positive Control”. Positive Control is a synthetic DNA which contains the target regions of the N1, N2 and RNaseP genes that are amplified with the primer-probe sets included in this kit. The positive control is used to identify possible qPCR inhibition and confirm the integrity of the reagents. It is not isolated from the virus, it does not have the entire virus genome and has no infectious properties. Positive controls represent a source of cross-contamination. Precautions should be taken to prevent and minimize the risk.

Primer/probe sets included in the POLGEN SARS-CoV-2 (COVID-19) Multiplex qPCR Detection Test are:

•                 The probe that detects the N1 region on COVID-19 is labelled with the FAM™️ fluorophore
•                 The probe that detects the N2 region on COVID-19 is labelled with the HEX™️ fluorophore
•                 The probe that detects the RNaseP is labelled with the Cy5™️ fluorophore

Primer-probe set of the POLGEN SARS-CoV-2 (COVID-19) Multiplex qPCR Detection Test is based on current sequence alignments of SARS-CoV-2. This allows for the RNA detection and differentiation of SARS-CoV-2. ERYIGIT will perform routine in silico analysis and post-market surveillance to ensure that in the case of new mutations in the virus during the outbreak, a proper update of the test will be performed, and customers will be properly notified.

The test has been developed to be used as a single one-step reverse transcription qPCR test for each patient sample, utilizing PCR amplification of the targets (N1, N2 and RNaseP) by FAM, HEX and Cy5 labelled probes. The reaction time is approximately 1 hour, it may differ slightly depending on the qPCR device used. The test contains internal controls that monitor the performance of the test.

The Reaction Mix also contains a propriety mix of Reverse Transcriptase and DNA Polymerase and the necessary buffer solutions and reagents such as nucleic acids in order to perform the qRT-PCR reaction. All the reagents are premixed for ease-of-use.