«ALSENSE FLU-SARS» Real-Time PCR Kit for the qualitative detection of SARS-CoV‑2, Influenza A and B
«ALSENSE FLU-SARS» Real-Time PCR Kit is designed to detect RNA of SARS-CoV‑2 coronavirus, Influenza A & B by single-step real-time polymerase chain reaction with fluorescent detection in nucleic acid preparations. The kit works with nucleic acids isolated from clinical samples (nasopharyngeal and oropharyngeal swab, bronchoalveolar lavage, endotracheal swab, sputum).
The kit is designed to operate with 96 samples. Aliquoting is possible. Resists 2–3 defrost-thaw cycles. The kit is intended for in vitro use only.
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Application field: Research practice for the qualitative RNA determination of SARS-CoV‑2 coronavirus, Influenza A and B.
User qualification requirements: The kit is intended for use by qualified personnel trained in molecular diagnostic techniques.
The test principle is to perform the reverse transcription reaction and the polymerase chain reaction sequentially in the same tube. The reverse transcription reaction is performed to form complementary DNA (cDNA) on the target RNA matrix. In the PCR step, the resulting cDNA is amplified multiple times. Detection of amplification products is performed during the analysis. For this purpose, special fluorescent tags (probes) are added to the reaction mixture, along with primers and other reaction components. Each DNA probe carries a fluorescent tag and a fluorescence quencher. When a specific product is formed, the DNA probe is destroyed and the quenching agent stops acting on the fluorescent tag, which leads to an increase in the fluorescence level.
The reaction mixture includes oligonucleotides and probes for simultaneous amplification and detection of RNA sites of viral genomes and the human genomic DNA RNase P gene (endogenous internal control). The use of endogenous internal control makes it possible to:
- evaluate the correctness of biomaterial collection, its transportation and storage;
- control the efficiency of nucleic acid isolation and PCR study steps, absence of reaction inhibition.
Amplification results are recorded using the following fluorescence detection channels (Table 1):
Table 1 – Detection channels for amplification products
|SARS-CoV‑2||Human RNase P gene||Influenza A||Influenza B|
«ALSENSE FLU-SARS» Real-Time PCR Kit (AL-SARSFLU-96, for 96 samples):
- Reaction buffer – 1,4 ml × 1 vial;
- Enzyme mixture – 100 µl × 1 vial;
- Positive control – 100 µl × 1 vial;
- Negative control – 500 µl × 1 vial.