«NOVAprep® 16/18 HPV» Real-Time PCR Kit
«NOVAprep® 16/18 HPV» Real-Time PCR Kit is designed for testing on the presence of HPV 16 and 18 DNA and their differentiation in clinical samples by real-time polymerase chain reaction with hybridization-fluorescent detection in nucleic acid preparations. The kit works with nucleic acids isolated from clinical samples (the recommended type of sample for laboratory confirmation of papillomavirus infection is the scraped epithelial cell material from the urogenital tract, urine, ejaculate, prostate fluid, biopsy material).
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The kit is intended for in vitro use only.
Application field: Application in clinical laboratory diagnostics and in research practice for qualitative DNA determination of human papillomaviruses of types 16 and 18 and their differentiation.
User qualification requirements: The kit is intended for use by qualified personnel trained in molecular diagnostic techniques.
During PCR, in the presence of genetic material of the papillomavirus infectious agent, the DNA fragment is multiply amplified. 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 that is detected by the amplifier.
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:
|HPV 16||Human RNase P gene||HPV 18|
«NOVAprep® 16/18 HPV» Real-Time PCR Kit (for 100 samples):
- Reaction buffer – 1,5 ml × 1 vial;
- Enzyme mixture – 0,12 ml × 1 vial;
- Positive control – 0,1 ml × 1 vial;
- Negative control – 0,5 ml × 1 vial.