On January 11, 2020, Chinese health authorities preliminarily identified more than 40 human infections with a novel coronavirus in an outbreak of pneumonia under investigation in Wuhan City, Hubei Province, China. The Chinese authorities identified a new type of coronavirus (novel coronavirus, named as 2019-nCoV), which was isolated on 7 January 2020.
Coronaviruses are a large family of viruses, some causing illness in humans and others circulating among animals such as camels, cats, and bats. 2019-nCoV is a novel coronavirus. The primer and probe design for this kit is based on the newly released strain (2019-nCoV) (GeneBank accession: MN908947) and covers 6 2019-nCoV strains sequences (EPI_ISL_402119, EPI_ISL_402120, EPI_ISL_402121, EPI_ISL_402122, EPI_ISL_402123, EPI_ISL_402124 included).
The kit contains a specific ready-to-use system for the detection of Novel Coronavirus (2019-nCoV) by Reverse transcription Polymerase Chain Reaction (RT-PCR) in the real-time PCR system. The master contains a Super Mix for the specific amplification of virus RNA. The reaction is done in one step real time RT-PCR. The first step is a reverse transcription (RT), during which the virus RNA is transcribed into cDNA. Afterwards, a thermostable DNA polymerase is used to amplify the specific gene fragments by means of polymerase chain reaction (PCR). Fluorescence is emitted and measured by the real time systems ́ optical unit during PCR. The detection of amplified virus DNA fragment is performed in fluorimeter channel FAM, HEX/VIC/JOE and Cal Red 610/ ROX/TEXAS RED with the fluorescent quencher BHQ1.
• All reagents should be stored at -20°C. Storage at +4°C is not recommended.
• All reagents can be used until the expiration date indicated on the kit label.
• Repeated thawing and freezing (> 3x) should be avoided, as this may reduce the sensitivity of the assay.
• Cool all reagents during the working steps.
• Super Mix should be stored in the dark.
Additionally Required Materials and Devices
- Biological cabinet
- Vortex mixer
- Sterile filter tips for micro pipets • Disposable gloves, powderless
- Real time PCR system
- Reak time PCR reaction tubes/plates
- Pipets (0.5μl - 1000μl)
- Sterile microtubes
- Biohazard waste container
- Tube racks