MOLgen - SARS-CoV-2 Real Time RT-PCR Kit

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MOLgen SARS-CoV-2 Real Time RT-PCR Kit is a multiplex one-step RT-PCR assay 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, salivary specimens 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 East 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-CoV-2 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 a multiplex real-time one-step RT-PCR assay 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.

The primer and probe set is designed to detect 4 targets for SARS-CoV-2: 3 specific for SARS-CoV-2 (N gene, S Gene, E gene) and 1 common for sabercoviridae (RdRp gene).

The detection of amplified virus RNA fragments is performed in fluorimeter channels: FAM (RdRp gene), ROX/Texas Red (E gene), Cy5.5/Alexa Fluor (S gene) and CY5 (N gene).

The use of human beta actin as Internal Control (IC) on the HEX channel monitoring correct amplification and cellularity 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), Gentier 96E (Xi’an Tianlong).

The kit contains reagents required for 100 reactions.

1. MOLgen SARS-CoV-2  Master Mix Ready to use reagent. Contains reagents for reverse transcription and amplification.

2. MOLgen SARS-CoV-2 Probe Mix Ready to use reagent. Contains primers and  probes which are premixed to the working concentrations to identifying 3 specific targets to SARS-CoV-2 (N , S and E genes) 1 common target for sabercoviridae (RdRp gene) and 1 for human beta actin.

3. MOLgen SARS-CoV-2 Positive Control Ready to use control. Contains  plasmids  with SARS-CoV-2 sequences (RdRP, E, S and N genes) plus an Internal Control (IC). The Internal Control is a plasmid with sequence of human beta actin.

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.

MATERIALS AND DEVICES REQUIRED BUT NOT SUPPLIED

• Nucleic Acid extraction kit intended for isolation of RNA (MOLgen Universal Extraction Kit (Adaltis) Ref. ME188830)
• Real-time PCR cycler
• Biosafety Cabinet
• Refrigerator
• Microcentrifuge with rotor for 1,5 mL reaction tubes
• 1.5 mL microcentrifuge tube for extraction
• Proteinase K Qiagen a ≥ 20mg/mL  - ≥ 600 U/mg
  • code  19131   2 mL/vial
  • code 19132  10 mL/vial
• Saliva collectors: Salivette® Sarstedt   code 51.1534   or sterile polipropilene wide mouth centrifuge tubes with conical bottom · Centrifuge
• Thermoblock
• Plate shaker
• Calibrated adjustable pipettes
• Disposable pipette tips with aerosol filter
• Strip Tubes and Caps (0.2 mL) or PCR Tubes (0.2 mL) or 96 well plate with optical caps or films
• Disposable medical non-sterile powder-free gloves
• Magnetic rack for tubes · Biohazard waste container

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.

STORAGE AND TRANSPORTATION

• Transport the kit at -15°C…-25°С.
  
• Store the kit in the manufacturer’s packaging at -15°C…-25°С for the entire shelf life.
  
• Do not freeze the Kit more than 3 times.