[Product Name]
Common name: COVID-19 Nucleic Acid Detection Kit(RT-NC-qPCR Fluorescence probe method)
[Packaging Specifications]
50 tests
[Intended Use]
This kit is suitable for the qualitative detection of novel coronavirus (2019-ncov) ORF1ab and N genes in throat swabs, bronchoalveolar lavage and plasma samples. Common signs of a person infected with a coronavirus include respiratory symptoms, fever, cough, shortness of breath, and dyspnea. In more severe cases, infection can cause pneumonia, severe acute respiratory syndrome (SARS), kidney failure, and even death. Coronavirus can be excreted through respiratory secretions or transmitted through oral fluids, sneezing, physical contact, and through air droplets.
[Inspection Principle]
This kit takes novel coronavirus (2019- nCoV) ORF1ab and the conserved sequence encoding the nucleocapsid protein N gene as the target region, and designs specific primer probes for dual target geneswhile covering the pol of COVID-19 strain ay27874 Gene region, and an internal reference gene was designed. RT-NC-qPCR technology is used to monitor PCR amplification in real time to achieve real-time monitoring of RT-qPCR and quantitative analysis of COVID-19.
[Main Components]
|
Component description
|
Quantity
|
|
Sample virus lysate
|
2.5 ml
|
|
Positive quality control product (pseudovirus)
|
50µl
|
|
RT-qPCR reaction mixture
|
50 tests
|
|
Primer probe mixture (P-M)
|
50 tests
|
[Storage Conditions and Shelf Life]
Stored at -20 ° C in dark condition, the validity period is 12 months. After opening the kit, store it at 2-8 ° C , and the validity period is 7 days.
[Applicable Instrument]
Real-time quantitative fluorescence PCR instrument that can simultaneously detect two kinds of fluorescence.
[Sample Requirements]
1. Applicable sample type: throat swab, bronchoalveolar lavage and plasma
2. Sample collection:
2.1 Throat swab: add 50ul virus lysate into a 1.5ml test tube, gently wipe the throat with throat swab, dip the head of the throat swab into the 1.5ml test tube for 2-3 seconds, and the throat swab is poured into 75% ethanol, and the test tube is tightly covered. Heat at 60 ° C for 5 minutes, then heat at 90 ° C for 10 minutes to obtain the RNA solution, and place it in ice for later use.
2.2 Plasma: take 1.5 ml test tube from 20ul plasma to 50ul virus lysate, cover tightly, heat at 60 ° C for 5 minutes and 90 ° C for 10 minutes, prepare RNA solution, and put it in ice for later use.
2.3 Bronchoalveolar lavage: take 50ul bronchoalveolar lavage solution in a 1.5ml test tube, mix with 50ul sample virus lysis solution, covered tightly, heat at 60 ° C for 5 minutes, then heat at 90 ° C for 10 minutes, to obtain RNA solution, and place on ice for later use.
[Test Method]
1. Preparation and sampling of reagents
Take out RT-qPCR reaction mixture and primer probe mixture (P-M) from the kit, and mix on a vortex after melting at room temperature, and centrifuge for several seconds before use.
Take N PCR reaction tubes (N = number of samples to be tested + negative quality control product (sample virus lysate) + positive quality control product)
The individual amplification system was prepared in the following table:
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Composition
|
Volume
|
|
RT-qPCR reaction mixture
|
5 ul
|
|
Primer probe mixture (P-M)
|
10 ul
|
|
RNA sample
|
5 ul
|
2. The PCR amplification
Put the reaction tube/plate in the fluorescence PCR instrument, and RT-NC-qPCR thermal cycling parameters were recommended:
|
Reaction
|
Phase
|
Cycles Number
|
Temperature
|
Holding time
|
|
Reverse transcription
|
1
|
1
|
42 ° C
|
10 min
|
|
Inactivated reverse transcriptase
|
2
|
1
|
95 ° C
|
3 min
|
|
PCR amplification
|
3
|
40
|
95 ° C |
10 s
|
|
46 ° C
|
30 s
|
|
60 ° C
|
30 s
|
Phase I: RT incubation at 42 ° C for 10 minutes
Phase II: 95 ° C3 minutes
Phase III: Amplification of 40 cycles: 95 ° C for 10 s, 46 ° C for 30 s, 60 ° C for 30 s, fluorescence signals of FAM and TET are detected at 60 ° C , FAM corresponding to the novel coronavirus specific orf1ab and N genes, and TET corresponding to the human ACE2 gene.
3. Quality control
Negative control: no obvious amplification curve
Positive control: there is an obvious amplification curve
The above requirements should be met in the same experiment, otherwise, the experiment will be invalid and need to be conducted again.
[Limitations of Test Results]
1. This kit is only used for clinical in vitro diagnosis and should be operated by personnel with professional experience or qualified training.
2. The test results of this kit are for clinical reference only. The clinical diagnosis of covid-19 should be considered in combination with its symptoms, signs, medical history, travel history and contact history with confirmed cases.
3. Personal protection should be done before the experiment to detect the potential infectivity of the samples.
4. Polluted waste shall be disposed of in accordance with regulations and must conform to national environmental, health and safety regulations.
5. Please read the instruction carefully before use and use it within the validity period.
6. The kit should be stored at -20 ° C in cold and dark, avoid repeated freeze-thaw. After thawing, it can be stored for 7 days at 2~8 ° C .
[Performance Indicators]
1. Repeatability: the positive quality control (pseudovirus) was repeatedly measured for 5 times, and the Ct value should be within the range of mean±3SD.
2. Sensitivity: the test was carried out after dilution of the positive quality control, and the minimum detection limit should be 500 copies/ml.
[Notes]
1. This reagent is only used for clinical in vitro diagnosis, and should be operated by personnel with professional experience or qualified training.
2. The test results of this reagent are for clinical reference only. The clinical diagnosis of the patient should be considered in combination with its symptoms, signs, history, other laboratory tests and treatment response.
3. Personal protection should be done before the experiment to detect the potential infectivity of the samples.
4. The reagent should be covered and preserved in time every time it is used, so as to prevent contamination, so as not to affect the dyeing effect.
5. Polluted waste shall be disposed of in accordance with regulations and must conform to national environmental, health and safety regulations.
6. Please read the instruction carefully before use and use it within the validity period.
7. Avoid high and low temperatures when storing reagents, and keep them out of direct sunlight.
8. Please refer to the packaging box for batch number, production date and expiry date.
[Symbols on label and package]
[Manufacturer]
Manufacturer: Aicon Biotech Inc
Contact information: +1 5034305188
Production address:8194 SW Durham Rd, Tigard, OR 97224 OR USA
After-sales service unit: Aicon biotech inc
Contact information: +1 5034305188
SW Durham Rd, Tigard, OR 97224 OR USA
Email address: Info@aiconbiotech.com
[EU Authorised Representative]
Osmunda Medical Technology Service GmbH
Site: Von oppen-weg 15, 14476 Potsdam Germany
DIMDI code: DE / 0000047267
