gb SARS-CoV-2 Variant E484K, L452R
one-step real-time PCR
Multiplex CE IVD kit enables detection of SARS-CoV-2 virus RNA and simultaneously the detection of mutations E484K and L452R in one reaction. The kit is based on one-step RT-qPCR using fluorescently labelled probes (FAM, HEX, ROX channels) and exogenous positive control (Cy5 channel).
– CE IVD one-step RT-qPCR kit
– multiplex SARS-CoV-2 detection of RdRP gene and mutations E484K, L452R in the S gene
– E484K is a key so-called “escape” mutation that causes reduced binding of neutralizing antibodies and is found in Beta (B.1.351, so-called South African) and Gamma (P.1, so-called Brazilian) variants
– L452R mutation is found in variants Delta (B.1.617.2, so-called Indian), Kappa (B.1.617.1, so-called Indian), Epsilon (B.1.427, B.1.429, so-called California)
– Assay CoV-2 E484K, L452R uses fluorescence channels FAM (mutation E484K), HEX (mutation L452R), ROX (gene RdRP), Cy5 (exogenous positive control EPC)
– kit further includes Master Mix OneStep Multi, Positive Control CoV-2 IND, EPC Template RNA and Deionized Water
– Positive Control CoV-2 IND serves for a verification of the analysis validity
– EPC Template RNA is an exogenous positive control for verification of the isolation process and one-step RT-qPCR reaction
– LOD of multiplex assay is 5.11 cop/rxn for RdRP gene, 25.46 cop/rxn for L452R mutation and 22.40 cop/rxn for E484K mutation (95% CI)
– the kit contains all the necessary components to perform the test
– analysis preparation scheme
Prevention of false negative results
In the kit, control of both RNA extraction step and PCR process is provided by detection of external RNA in Cy5 channel after spiking the samples with the EPC Template RNA prior to the extraction.
In addition to this generally used protocol, the labs that demand a verification of the sampling step can use our gb Human B2M mRNA kit. The kit is based on the detection of transcripts of B2M gene, a human reference gene ubiquitously present in all human samples. If the human reference transcript is not detected, the sample was not taken properly.