Kemogen Covid-19 qPCR Diagnostic Kit

Diagnostics » Genetic

Kemogen Covid 19 qPCR diagnostic kit is a multiplex-based real-time qPCR kit to detect the presence of relevant target regions of SARS-CoV-2 RNA in respiratory tract samples such as nasopharyngeal, oropharyngeal, sputum and bronchoalveolar lavage. The kit contains labeled oligonucleotides designed specifically for the "N" and "Orf1ab" genes specific to SARS-CoV-2 and the human "RNaseP" (Ribonuclease P) gene as a positive control. Kemogen Covid 19 diagnostic kit performs cDNA synthesis and qPCR reaction for the detection of SARS-CoV-2 RNA in the same tube.

KIT STORAGE CONDITIONS
All kit contents should be stored in the dark between -15 o C and -25 o C. The kit should be consumed before the expiration date written on the box. If the kit content is exposed to more than 5 freezing-thawing processes, the sensitivity of the kit may be impaired, stability and quality are not guaranteed.

DEVICES COMPATIBLE WITH THE KIT
Kemogen Covid 19 qPCR diagnostic kit is compatible with BioRad CFX96, Qiagen Rotor-GeneQ, LightCycler 480 (Roche), ABI Prism StepOne Plus devices with FAM and HEX channels. The kit does not contain ROX dye. For this reason, the ROX option should be turned off in devices that require the use of passive dye (ABI Prism StepOne Plus).

PROTOCOL SAMPLE PREPARATION
The protocols of commercial kits used for RNA isolation from respiratory tract samples should be applied and the resulting RNA should be used as a direct template nucleic acid for the reaction.

The kit contents should be thawed on ice before use. The tubes can be mixed by briefly and gently vortexing. Mixtures of reactants should be prepared under the conditions specified below, including one positive and one negative control for each run, in accordance with the number of samples. The prepared mixture should be aliquoted as 15 μl into PCR tubes suitable for the relevant devices, and 5 μl of the RNA isolated from the samples should be added to it. All work must be carried out on a cold plate. Kit contents and reaction mixtures should not be exposed to direct light for a long time.

DATA ANALYSIS BIORAD DEVICE
Automatic threshold value should be used for FAM and HEX.

ROTORGENEQ DEVICE

1. “New run wizard” tab
2. On the screen that opens with “Gain optimization”
3. “Green” and “Yellow” should be added in the channel list.
4. If not, default values ​​should be added with the "Optimized acquiring" option. The “Perform optimization before 1st acquisition” option must also be selected. After the reaction, analyzes should be performed by following the following steps: 1. "Quantification analysis" should be performed with the "Cycling Green" and "Cycling Yellow" tabs.
5. “Dynamic tube” should be selected.
6. “Ignore first” should be set to “10”.
7. “Outlier removal” should be set to “5%”.
8. The “threshold” in the “CT Calculation” tab should be set to “0.03” and “Eliminate cycles before” should be set to “0”.