Objectives . Increased importance in detection and surveillance of SARS-CoV-2 has been demonstrated due to the emergence of variants of concern (VOCs). In this study we evaluated if a commercially available real-time SARS-CoV-2 PCR assay can identify B.1.1.7 lineage samples by a specific N gene dropout or Ct value shift compared to the S or RdRP gene . Methods . Patients samples with confirmed B.1.1.7 variant by whole-genome sequencing and variant-specific PCR (n=48) and non-B.1.1.7 samples (n=53) were tested by the Allplex SARS-CoV-2/FluA/FluB/RSV PCR assay for presence of S, RdRP and N gene of SARS CoV-2 . The N gene coding sequence of SARS-CoV-2 with and without D3L mutation (specific for B.1.1.7) were cloned into pCR-TOPO vectors and Allplex SARS-CoV-2/FluA/FluB/RSV PCR assay was performed . Results . All studied B.1.1.7 patient samples showed significantly higher Ct values (delta 6-10, N-gene dropout on Ct values> 29) in the N gene compared to the respective values of S and RdRP gene . Receiver operating characteristic (ROC) curve analysis resulted in 100% sensitivity and specificity for delta Ct N/RdRP and delta Ct N/S . As a result of the reversed genetic experiments we found also the shift in Ct values for the 3L variant N-gene . Conclusions . N gene dropout or Ct value shift is specific for B.1.1.7 positive samples using the Allplex SARS-CoV-2/FluA/FluB/RSV PCR assay . This approach can be used as a rapid tool for B.1.1.7 detection in single assay high throughput diagnostics.