Influenza A H1 + H3 Seasonal PCR Kit CE/IVD
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RealCycler HIFL-
Catalog Number: RealCycler HIFL-
1 Intended use:
RealCycler HIFL is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of Influenza A H1N1 pdm09, Seasonal Influenza A H1 and Seasonal Influenza A H3 in clinical samples.
The system includes an internal control CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition.
2 Principle of the test
The polymerase chain reaction (PCR) is based on the amplification of a specific region of the DNA/RNA by using complementary primers to the target sequence. Real-time PCR uses marked probes with fluorophores that emit fluorescence in the case of amplification. The cycle of the PCR protocol in which appears significant fluorescence is proportional to the DNA/RNA quantity present in the sample. This value is called Cycle Threshold (Ct) or Cycle Quantification (Cq).
In the HINI amplification, H1N1 pdm09 is detected in the corresponding channel of FAM fluorophore and the internal control is detected in the corresponding channel of Alx532 or HEX fluorophore.
In the FLUS amplification, Influenza A H1 is detected in the corresponding channel of TxR fluorophore, Influenza A H3 is detected in the corresponding channel of Alx647 fluorophore and the internal control is detected in the corresponding channel of Alx532 or HEX fluorophore.
3 Technical specifications
Sensitivity:
-Influenza A H1N1 pdm09: 500 copies/µL.
-Seasonal Influenza A H1: 500 copies/µL.
-Seasonal Influenza A H3: 10 copies/µL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity:
- Influenza A H1N1 pdm09: hemagglutinin gene.
- Seasonal Influenza A H1: hemagglutinin gene.
- Seasonal Influenza A H3: hemagglutinin gene.
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
This technical specifications has been verified using QCMD 2011 Influenza virus Haemagglutinin Typing.
4 Contents
RealCycler HIFL includes the HINI AmpliMix and FLUS AmpliMix, a retrotranscriptase, a FluA H1N1 RNA Positive Control and FLUS RNA Positive Control which contains a RNA mixture of Seasonal Influenza A H1 and H3.
All reagents are ready to use without adding or rebuilding any component.
RealCycler HIFL is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of Influenza A H1N1 pdm09, Seasonal Influenza A H1 and Seasonal Influenza A H3 in clinical samples.
The system includes an internal control CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition.
2 Principle of the test
The polymerase chain reaction (PCR) is based on the amplification of a specific region of the DNA/RNA by using complementary primers to the target sequence. Real-time PCR uses marked probes with fluorophores that emit fluorescence in the case of amplification. The cycle of the PCR protocol in which appears significant fluorescence is proportional to the DNA/RNA quantity present in the sample. This value is called Cycle Threshold (Ct) or Cycle Quantification (Cq).
In the HINI amplification, H1N1 pdm09 is detected in the corresponding channel of FAM fluorophore and the internal control is detected in the corresponding channel of Alx532 or HEX fluorophore.
In the FLUS amplification, Influenza A H1 is detected in the corresponding channel of TxR fluorophore, Influenza A H3 is detected in the corresponding channel of Alx647 fluorophore and the internal control is detected in the corresponding channel of Alx532 or HEX fluorophore.
3 Technical specifications
Sensitivity:
-Influenza A H1N1 pdm09: 500 copies/µL.
-Seasonal Influenza A H1: 500 copies/µL.
-Seasonal Influenza A H3: 10 copies/µL.
The analytical sensitivity has been determined by limit dilution. This sensitivity has been showed in repeated assays with reproducibility over 95%.
Specificity:
- Influenza A H1N1 pdm09: hemagglutinin gene.
- Seasonal Influenza A H1: hemagglutinin gene.
- Seasonal Influenza A H3: hemagglutinin gene.
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
This technical specifications has been verified using QCMD 2011 Influenza virus Haemagglutinin Typing.
4 Contents
RealCycler HIFL includes the HINI AmpliMix and FLUS AmpliMix, a retrotranscriptase, a FluA H1N1 RNA Positive Control and FLUS RNA Positive Control which contains a RNA mixture of Seasonal Influenza A H1 and H3.
All reagents are ready to use without adding or rebuilding any component.
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