1. Specimen collection: collect samples of the upper respiratory tract (including nasopharynx and oropharynx) and lower respiratory tract (including sputum, tracheal aspirates or bronchoalveolar lavage fluid) at the same time, and perform nCoV detection by RT-PCR. In cases where lower respiratory tract samples are readily available: use appropriate PPE for specimen collection (note URT specimens for droplets and exposure precautions, and LRT specimens for air prevention) When collecting URT samples, use a virus sampling cotton swab (sterile polyester or rayon instead of cotton) and virus transport medium. Do not sample from nostrils or tonsils. For patients suspected of having nCoV, especially patients with pneumonia or severe disease, a single URT sample cannot exclude the diagnosis, it is recommended to increase URT and LRT samples. LRT (relative to URT) samples are more likely to be positive, and as stated, in cases where lower respiratory tract samples are readily available (eg, mechanically ventilated patients), clinicians may choose to collect only lower respiratory tract samples).

2. Nucleic acid extraction: nucleic acid extraction instrument or manual nucleic acid extraction

1), without instruments, use manual extraction

Equipment and lab consumables required for nucleic acid extraction and sample separation:

Biological safety cabinet (function: separation of samples, provision of personnel, samples and environmental protection),

Desktop high-speed refrigerated centrifuge (function: speed ≥ 12000RPM, analysis of nucleic acid, etc.),

Low temperature refrigerator (function: reagent storage),

Metal bath, pipette (function: precise sample addition),

Sterilization centrifuge tube (function: separation of nucleic acids, etc.),

Latex gloves (function: anti-pollution),

Absolute ethanol, isopropyl alcohol, DNase and RNase-free water (non-DEPC treated water).

3. Automatic nucleic acid extraction instrument: (automatic extraction, 32 samples can be extracted in 26 minutes at a time)

The supporting pre-packing plate is directly added with samples and directly loaded onto the machine for extraction:

4. Fluorescence quantitative PCR detection:

Required equipment and consumables: biological safety cabinet (function: sample preparation, provision of personnel, samples and environmental protection),

Fluorescence quantitative PCR instrument (function: PCR amplification and detection analysis),

Desktop high-speed refrigerated centrifuge (function: speed ≥ 12000RPM),

Low temperature refrigerator (-86 ℃ sample storage, -20 ℃ and 4 ℃ reagent storage),

Oscillator, (mix)

Graduated Pipette Tip (Function: Precise loading), 1-10ul specification

Sterilization centrifuge tube (0.5 / 1.5 / 2ml),

0.2ml optical grade PCR tube / plate (function: PCR reaction tube),

Latex gloves (function: anti-pollution).

Nucleic acid extraction, reaction system configuration, result interpretation, etc. are subject to the kit instructions.