Adenovirus Rapid Test-Nasal Secretion

Intended Use:

SARS-CoV-2 Nucleic Acid Test kit is apply to the qualitative detection of nucleic acid of COVID-19 in nasopharyngeal swabs, oropharyngeal swabs and sputum. The test results provide molecular diagnosis basis for infection or suspected patients, and should not be used as the sole criterion for clinical diagnosis.

Materials:

Test Procedure:

A. Sample Preparation

Extracted RNA is the starting material for the SARS-CoV-2 Nucleic Acid Test kit.

Important: Each nucleic acid extraction procedure must be performed simultaneously with one Positive control(40 μL, dilute with sterile saline solutions/PBS to desired volume) and one Negative control(40 μL, dilute with sterile saline solutions/PBS to desired volume).

It is recommended to use the the following commercial nucleic acid extraction kits: PureLinkTM Viral RNA/DNA Mini Kit (Invitrogen); Viral RNA Mini kit (QIAGEN); TIANamp Hi-DNA/RNA Kit (TIANGEN);

B. Reaction Mix setup

1. All reagents and samples should be thawed completely, mixed completely and centrifuged briefly at room temperature (approximate range 18 to 25 °C) before use.

2. Prepare the required volume of the Reaction Mix in an appropriately sized polypropylene microcentrifuge tube.

3. Pipette 25 µL of the Reaction Mix into each required well of an appropriate optical 96-well reaction plate or an appropriate optical reaction tube.

C. Template addition

1. Add 5 μL of extracted RNA from Samples/Positive control/Negative control. The total volume is 30 μL.

2. Thoroughly mix the samples and controls with the Reaction Mix by pipetting up and down/Vortex.

3. Close the 96-well reaction plate with appropriate lids or optical adhesive film and the reaction tubes with appropriate lids.

4. Centrifuge the 96-well reaction plate in a centrifuge with a microtiter plate rotor for 30 seconds at approximately 2500rpm.

D. Programming the Real-Time PCR Instrument

For basic information regarding the setup and programming of the different real-time PCR instruments, please refer to the user manual of the respective instrument.The assay has been optimized for the ABI7500 Real-Time PCR system. The PCR run is performed with cycling conditions as described in the table below.

Define the fluorescence detectors

*Please set the fluorescence internal reference of the instrument to "None", for example: For ABI series instruments set "Passive Reference" to "None".

E. Data analysis

Please set the analysis parameters and analyze the data according to the operating instructions of the relevant instruments.

Take ABI 7500 as an example:

The Baseline should be set to a range that eliminates the background fluorescence found in the early cycles of amplification, but which does not overlap the area where amplifications signals rise above the background(Start value:3~10; End value: 10-20). The Cycle Threshold (Ct) should be in the base of the exponential phase. Click "Analysis" to obtain the analysis result automatically, and read the detection result in the "Report" window.

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