SARS-CoV-2 VoC qPCR Assay N501Y del69-70

Simultaneous SARS-CoV-2 & VoC-PCR for Variants of Concern B.1.1.7, B.1.351 and P1

What are Variants of Concern ?

Strains with a significant impact on transmissibility, severity and/or immunity that are likely to aggravate the epidemiological situation in the EU/EEA are considered as Variants of Concern (VoC).

Currently, four variants are considered as VoC: P1 (“Brazilian”), B.1351 (“South African”), P.1.1.17 (“British”) and the P.1.617 (“Indian”). All VoC have specific mutations for which we have designed highly specific qPCR assays for a clear distinction of all VoC.

Besides the M-gene, our assay specifically detects the N501Y mutation present in all VoC (B.1.1.7; B.1.351 and P1). In addition, the assay is specific for the S gene (wt) and detects the del69-70 by drop-out. This way, our assay detects both wt or the Indian Variant and VoC via at least two targets (dual target assay). In contrast to some other assays, which lead to a complete drop-out of signals our assay will always generate at least two positive signals.

Our assay also includes RNaseP as sampling control. The assay can be performed on common qPCR instruments with detection channels of FAM, Joe, Cy5, and ROX.

Only one qPCR reaction is needed for fastest results.

For the identification of the B.1.617 (“Indian”) variant, we recommend our B.1.617-specific assay when N501Y and del69-70 are negative.

The assay is for research use only at the moment. CE-IVD is in progress.

Left: WT leading to a S-gene signal but no signal with the N501Y specific assay. Right: B1.1.1.7 strain with an N501Y specific signal but no signal for the S-gene because of D69/70 (drop out). Both strains lead to speficic PCR product signals for the SARS-CoV-2 M gene and the RP-gene (human control gene).