Description:
This kit is used for the in vitro qualitative detection of novel coronavirus (2019-nCoV) ORFlab and N gene in the throat swabs, sputum specimens of suspected pneumonia patients infected by novel coronavirus, suspected clustering cases and others needing diagnosis or differential diagnosis for novel coronavirus.
Detection of novel coronavirus RNA shall meet the requirements of related guideline, clinical diagnostic/disease control and other documents, so as to carry out the regulations on biosafety. The detection results of this kit are for clinical reference only and should not be used as the sole criteria for clinical diagnosis. It is recommended to conduct a comprehensive analysis on the condition in combination with the clinical manifestations of the patient and other laboratory tests.
Test Principle:
This kit is based on one-step RT-PCR technique. In practice, 2019 Novel Coronavirus (2019-nCoV) ORF1ab and N genes are selected as amplification target regions. Specific primers and fluorescent probes are designed for the detection of 2019 Novel Coronavirus RNA in the specimens. This kit also includes an endogenous internal standard detection system, which is used for monitoring over the processes of specimen collection, RNA and PCR amplification, thereby reducing false negative results.

Components:

The above components in different batches of kits cannot be interchangeable.
Reagents to be self-prepared: RNA extraction or purification reagents. It is recommended to use YHXB No. 20170583 and YHXB No. 20150302 products manufactured by DAAN.
Description of negative/positive control materials: the positive control material is pseudovirus containing the target fragments and pseudovirus containing internal standard fragments, while the negative control material is pseudovirus containing an internal standard fragments. During use, they should be involved in extraction and should be considered as an infectious substance. They shall be handled and disposed in accordance with relevant regulations.

Storage Conditions and Shelf life:
The kit is stored at -20 ± 5°C, and the validity period is 6 months.
The reagent can be stored at 4°C for 3 days; the kit can be stored at 37°C for 3 days; repeated freezing and thawing should be avoided; repeated freezing and thawing cycles should not exceed 7 times and the times of decapping the reagent should not exceed 7.
See the product label for the date of manufacture and validity of the kit.

Applicable Instruments:
ABI 7500, LightCycler480, AGS4800, Bio-Rad CFX96.

Specimen Requirements:
1. Applicable sample types: throat swabs and sputum.
2. Specimen collection (aseptic technique)

Throat swab: Wipe the tonsil and posterior pharyngeal wall with two swabs at the same time, and immerse the swab heads into the tube containing the sampling liquid;
Sputum: After the patient has a deep cough, collect the coughed sputum in the screw-cap test tube containing the sampling liquid.

Bronchoalveolar lavage fluid (BALF): After local anesthesia, insert the bronchoscope through pharyngeal into bronchus of middle lobe of right lung or lingual segment of left lung via mouth or nose, place the top into the branch opening of bronchus, add sterile saline slowly via trachea biopsy hole, 30-50ml per time, volume 100-250ml, not exceed 300ml.

Anus swab: Insert disinfect cotton swab into anus 3-5cm, pull out after rotation, put into virus preservation solution collection tube immediately, discard the end of swab, screw the collection tube.

Blood sample: Collect blood sample 5ml recommended using EDTA vacuum tube, confirm using whole blood or plasma to perform nucleic acid extraction according to the requirement of nucleic acid kit. If separating plasma is needed, centrifuge the whole blood at 1500-2000rpm for 10 minutes, collect supernatant into sterile collection tube.

3. Sample storage and transportation

The specimens for virus isolation and RNA detection should be tested as soon as possible. The specimens that can be detected within 24 hours can be stored at 4 °C; those that cannot be detected within 24 hours should be stored at -70°C or below (if no -70°C storage conditions, then temporarily stored in -20 °C refrigerator). The specimens should avoid repeated freezing and thawing during transport. Specimens should be sent to the laboratory as soon as possible after collection. If the specimens need to be transported over long distances, it is recommended to use dry ice for storage.

Reference
1. Clinical management of severe acute respiratory infection when novel coronavirus (nCoV) infection is suspected Interim guidance. 2020.