Coronaviruses are a family of large RNA viruses with size ranging from 26 to 32 kb. These viruses are zoonotic and in humans can cause respiratory infections. As the coronavirus is an RNA virus, it has a relatively high mutation rate resulting in rapid evolution. In December 2019, a new deadly coronavirus known as SARS-CoV-2, which has a high sequence similarity to SARS-CoV, was identified as the cause of the Covid-19 outbreak. Since then, numerous variants of SARS-CoV-2 were reported around the world. Among them, one mutation was detected within the Spike protein of SARS-CoV-2 and is known as the spike-N501Y mutation. This variant causes the amino acid asparagine at position 501 (N501) mutate to a tyrosine (Y501), and it may lead to a much higher infection rate. The spike-N501Y mutation is thought to contribute to the higher viral binding affinity to the ACE2 receptor on the human cell membrane. This mutation is present in some major circulating SARS-CoV-2 strains including B.1.1.7, 501Y.V2, and P.1.

ScienCell's SARS-CoV-2 N501Y Mutation Multiplex RT-qPCR Screening Kit (SCVNY) is designed to screen for the presence of the SARS-CoV-2 spike-N501Y mutation. Two multiplex primer/probe set components (Cat #7128-REF and #7128-N501Y) are included in the kit. The reference primer/probe set component (Cat #7128-REF) contains 3 primer/probe sets, N1-FAM, N2-FAM, and RP-HEX (Table 1). Among them, N1-FAM and N2-FAM target two regions on the coronavirus SARS-CoV-2 nucleocapsid (N) gene. RP-HEX targets the exon 1 of human RPP30 gene and serves as a control to assess specimen quality. The N501Y primer/probe set component (Cat #7128-N501Y) contains 2 primer/probe sets, Y501-FAM and N501-HEX (Table 2), which target the coronavirus SARS-CoV-2 spike (S) gene with a tyrosine (Y) and an asparagine (N) at position 501, respectively. For more efficient screening, if the expected N501Y mutation rate is low, a pool of up to 10 RNA samples can be used as the template for one qPCR reaction. If results for Y501-FAM are negative, then all pooled samples do not contain the N501Y mutation. For Y501-FAM positive pooled samples, the pooled samples should be tested individually to identify the N501Y mutated one(s). Please refer to Tables 6 and 7 for results interpretation.