GENERAL INFORMATION
Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) is the name given to the new coronavirus that originated in 2019 and is the causative agent of COVID-19 (the disease caused by the SARS-CoV-2 virus). The virus was originally known as the 2019-nCoV nomenclature. SARS-CoV-2 is a new coronavirus strain that has not been previously identified in humans. The genome of these viruses consists of single-stranded RNA protected by two protein coatings - nucleocapsid and capsid. Its genetic characteristics are significantly different from SARS-CoV and MERS-CoV.
The SureFast® SARS-CoV-2 PLUS is a real-time RT-PCR for the direct, qualitative detection of novel coronavirus (SARS-CoV-2) RNA. The test kit enables the detection of the E-Gene, which encodes an envelope protein. Each reaction contains an Internal Control RNA (ICR, consisting of MS2-bacteriophage) as an internal control of sample preparation procedure and to monitor possible PCR-inhibition.
For RNA-preparation the use of SureFast® PREP DNA/RNA Virus (Art. No. F1051) is recommended.
The real-time RT-PCR assay can be performed with commonly used real-time PCR instruments, equipped for detection of two fluorescence emissions at the channels FAM and VIC/HEX at the same time. The technical validation of instruments was performed on Roche LightCycler® 480 II, Qiagen Rotor-Gene Q, Applied Biosystems 7500 Fast Dx, Bio-Rad CFX96, R-Biopharm RIDA®CYCLER, Agilent AriaDx and Agilent Mx3005P.
Kit components and storage:
Store all reagents at –20°C and protected from light.
Interpretation of results
SARS-CoV-2 RNA is detected in the FAM-channel. In the VIC/HEX-channel the internal amplification or positive extraction control (ICR) is detected. High amplicon concentrations can result in a weak or absent signal of the internal amplification control (IAC). A Cp value for the internal amplification control (IAC) is not needed to obtain a positive result of the positive control. A sample is stated positive for SARS-CoV-2, if the sample RNA shows amplification in the FAM-channel. A sample is stated negative for SARS-CoV-2, if the sample RNA shows no amplification in the FAM-channel and if the internal amplification or positive extraction control (VIC/HEX-channel) of the sample is positive with a shift in Cp-Value ≤ 2 compared to the negative control.
If the sample RNA in the VIC/HEX-Channel shows no amplification or a shift in Cp-value > 2 compared to the negative control, it contains PCR inhibiting substances. A significant decrease in the fluorescence signal can also show the presence of PCR inhibiting substances. Under these circumstances RNA isolation and purification of the sample need to be improved.
Test format: 100 reactions
LOD (Detection limit): < 25 RNA copies.
Note:
Samples tested negative with SureFast®SARS-CoV-2 PLUS still could contain a SARS-CoV-2 contamination below the limit of detection of the assay.