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|Line||Method & Sample||Product||Package Info|
|MOLgen||Nasopharyngeal swabs,Oropharyngeal swabs,Bronchoalveolar lavage fluid (BALF)||SARS-CoV-2 Real Time RT-PCR Kit||Tests per Package: 100|
|Respiratory||Detection of SARS-CoV-2 Virus||Code: MESARS-CoV-2||Package Format: Liquid Reagents|
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MOLgen SARS-CoV-2 Real Time RT-PCR Kit is used for the qualitative detection of Novel Coronavirus (SARS-CoV-2), by Reverse Transcription (RT) and Real Time Polymerase Chain Reaction (PCR) from RNA extracted from human respiratory specimens such as nasopharyngeal swabs, oropharyngeal swabs, sputum and bronchoalveolar lavage fluid (BALF).
Coronaviruses are a large family of respiratory viruses, some causing illness in human and others circulating among animals such as camels, cats and bats from common cold to severe diseases such as Middle Est Respiratory Syndrome (MERS-CoV) and Severe Acute Respiratory Syndrome (SARS-CoV).
SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) is a new strain of coronavirus emerged from Wuhan, China in December 2019.
The infection with SARS CoV2 virus causes Coronavirus disease (COVID-19) where "CO" stands for crown, "VI" for viruses, "D" for disease and “19'" indicates the year in which it occurred.
COVID-19 symptoms are fever, cough , pneumonia and Severe Acute Respiratory Syndrome (SARS).
MOLgen SARS-CoV-2 Real Time RT-PCR Kit is intended for the detection of the New Coronavirus SARS-CoV-2 RNA using reverse transcription of viral RNA and real-time polymerase chain reaction (RT-PCR) method with fluorescence detection of amplified product.
Real-time PCR is based on the detection of the fluorescence produced by a reporter molecule, which increases as the reaction proceeds.
Reporter molecule is a dual-labeled DNA-probe that specifically binds to the target region of the pathogen’s cDNA. Fluorescence signal increases due to the separation of fluorescence dye and quencher owing to Taq DNA-polymerase exonuclease activity during amplification. PCR consists of repeated cycles: temperature denaturation of cDNA, primer annealing and complementary chain synthesis.
Threshold cycle value (Ct) is a cycle number, at which the fluorescence generated within a reaction crosses the threshold and the fluorescence signal rises significantly above the background. The increased signal is due to the use of a DNA hybridization probe that is specific for the given cDNA sequence: it binds to the cDNA in the course of reaction and provides additional specificity of the method. The DNA probe consists of a fluorescence dye at the 5’-end and a fluorescence quencher at the 3’-end that significantly reduces the fluorescence intensity. During the polymerase synthesis of the complementary strand, the probe is cleaved from the 5’-end due to the 5’-3’ nuclease activity of Taq DNA polymerase, the quencher and the dye become separated thus increasing the fluorescence signal due to accumulation of the reaction product.
The detected fluorescence intensity depends on the initial quantity of pathogen’s cDNA template in the sample.
The primer and probe set is designed to detect SARSCoV2 N gene, E gene and RdRP gene.
The detection of amplified virus RNA fragment is performed in fluorimeter channel FAM, ROX and CY5. The use of Internal Control (IC) on HEX channel ensure the monitoring correct amplification of the samples.
The results of PCR analysis are taken into account in complex diagnostics of disease. The kit is validated for use with: AMPLIlab (Adaltis), CFX96 (Bio-Rad), RotorGene Q (Qiagen).
The kit contains reagents required for 100 reactions.
|Component Description||Nr Vial||Volume
|MESARSCoV2/MM||MOLgen SARS-CoV-2 Master Mix||1||1000||Green|
|MESARSCoV2/PM||MOLgen SARS-CoV-2 Probe Mix||1||500||Yellow|
|MESARSCoV2/PC||MOLgen SARS-CoV-2 Positive Control||1||50||Red|
|MESARSCoV2/NC||MOLgen SARS-CoV-2 Negative Control||1||50||Clear|
|Number of reactions (rxns)||1||100|
|Reference Kit Code||1||MESARS-CoV-2|
1. MOLgen SARS-CoV-2 Master Mix
Ready to use reagent.
It contains reagents for reverse transcription and amplification.
2. MOLgen SARS-CoV-2 Probe Mix
Ready to use reagent.
It contains primers and probes which are premixed to the working concentrations.
3. MOLgen SARS-CoV-2 Positive Control
Ready to use control.
It contains plasmids with SARS-CoV2 sequences (RdRP, E and N genes) plus an Internal Control (IC).
4. MOLgen SARS-CoV-2 Negative Control
Ready to use control.
Negative Control should be included at every run as it indicates that reagents have not been contaminated.
E. MATERIALS AND DEVICES REQUIRED BUT NOT SUPPLIED
N. STORAGE AND TRANSPORTATION
NOTE: The extraction protocol with MOLgen Universal Extraction Kit (Adaltis) Ref. ME188830 and the PCR set-up procedure can be done in automated mode using the Adaltis EXTRAlab Instrument by executing the extraction and PCR set-up protocol preloaded in the instrument user interface