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Download All Background: Coronaviruses are positive single-stranded RNA viruses of the family Coronaviridae. Several different strains of coronaviruses are currently known to infect humans (HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1, MERS-CoV, SARS-CoV, SARS-CoV-2, NCoV and HCoV-EMC). Strains HCoV-229E, HCoV-NL63, HCoV-OC43, MERS-CoV and HCoV-HKU1 cause cold, upper respiratory infection, bronchiolitis and pneumonia in humans. SARS-CoV, a beta coronavirus, causes the Severe Acute Respiratory Syndrome (SARS).
Description: ViroReal® Kit SARS-CoV-2 Multiplex (Version 2.0) is a non-automated IVD test, based on one-step reverse transcription real-time PCR (RT-PCR), for the qualitative detection of RNA of the N gene of SARS-CoV-2, SARS-CoV and SARS-related coronavirus (Sarbecovirus) and of the RdRp gene of SARS-CoV-2. Proper specimens are RNA extracts of samples from the human respiratory tract (throat rinsing fluid, nasopharyngeal and oropharyngeal swabs, anterior nasal swab and mid-turbinate nasal swab specimens, nasopharyngeal wash/aspirate and nasal aspirates). This test is suitable for patients of all ages with or without a suspected SARS-CoV infection and is intended as an aid in the diagnosis of SARS-CoV infection in combination with patient history and additional clinical information. The test is for professional use only and the use is limited to qualified personnel instructed in real-time PCR and in vitro diagnostic procedures. A probe-specific amplification-curve in the FAM channel (530 nm) indicates the amplification of the N gene specific RNA. A probe-specific amplification-curve in the VIC channel (554 nm) indicates the amplification of RdRp gene specific RNA. An internal RNA positive control (RNA IPC) is detected in Cy5 channel and is used as RNA extraction as well as RT-PCR inhibition control. The target for the RNA IPC (artificial target RNA) is added during extraction of the sample. The multiplex approach allows universal detection of all so far known SARS coronavirus strains (Sarbecovirus) including SARS-CoV-2, and the discrimination between SARS-CoV-2 and the other Sarbecovirus strains. The chosen design for the N gene includes a highly conserved region in all SARS coronavirus clusters of the N gene, which allows the universal detection of previously known Sarbecoviruses including SARS-CoV-2. Due to its universal character, it takes into account possible future mutations in the SARS-CoV-2 virus sequence. Because of its design, the test has the potential to detect new SARS corona viruses in the future. For the RdRp gene, a sequence range highly specific for SARS-CoV-2 was chosen, which allows the specific detection of SARS-CoV-2. The primer and probe designs chosen are not identical with designs published by the WHO.
| Order Number | Reactions | Channel Pathogen | Channel IPC | Target |
|---|---|---|---|---|
| DHUV02413 | 100 | FAM, VIC | Cy5 | N gene & RdRp |
| DHUV02413x5 | 500 | FAM, VIC | Cy5 | N gene & RdRp |
- Amplification and detection: N gene of SARS-CoV and RdRp gene of SARS-CoV-2
- Transcription with thermostable MMLV Reverse Transcriptase (M-MLV)
- contains RNase Inhibitor to block RNA degradation
- Real-time PCR with rapid hot-start Taq DNA polymerase
- ROX™ dye as passive reference
- Internal Positive Control System to exclude false-negative results
- Optimized to handle PCR inhibitors
- PCR- platforms: runs on all established standard real-time PCR- platforms
- Harmonized thermal profiles to run RNA and DNA samples simultaneously
PCR platforms: This test has been validated with the ABI® 7500 instrument (fast cycle parameters are not supported, Thermo Fisher Scientific) and was also tested with QuantStudioTM 7 Pro (Thermo Fisher Scientific), MIC instrument (bio molecular systems), Mx3005P® (Agilent), qTOWER3G (Analytik Jena) and LightCycler® 480 II (Roche Diagnostics). It is also compatible with other real-time PCR instruments which detect and differentiate fluorescence in FAM, VIC and Cy5 channel (eg., QuantStudio™ 5 (Thermo Fisher Scientific), cobas z 480 Analyzer (Roche),). When using PCR-platforms not tested by ingenetix, an evaluation of the multiplex-PCR shall be done. Keep in mind that some PCR-platforms first have to be calibrated with the corresponding dye before performing multiplex-PCR.