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General description
Test Principle:
The Amplifluor ID Influenza A, H5N1 Virus Reagent was designed to detect conserved nucleic acid regions contained within Influenza A virus genomes that encode for the hemagglutinin (H5) and neuraminidase (N1) genes. Since the Since the Infleunza A Virus (H5N1) contains an RNA genome, initial amplification reactions are performed using the reverse transcription (RT) primer for 1st strand cDNA synthesis, followed by Polymerase Chain Reaction (PCR) amplification utilizing the forward Amplifluor hairpin primer and reverse primer to produce target-specific fluorescent amplicons. The Amplifluor hairpin primer is labeled with Donor (fluorescein) and Quencher (DABSYL) fluorophores, which produce the high signal to noise ratio observed in the Amplifluor assay. Upon incorporation into new amplicons, the Amplifluor primer is fluorescent (reported in the instrument′s FAM layer), while the fluorescein signal of the unincorporated Amplifluor primer is quenched. When performed on a thermal cycler capable of real-time fluorescence detection, the amplification products of the PCR reaction can be monitored directly in a quantitative fashion, allowing evaluation of template detection in a single, closed-tube system with no additional assay procedure. Where required, template standard curves may be employed to quantitatively determine viral load.
The Amplifluor ID Influenza A, H5N1 Virus Reagent was designed to detect conserved nucleic acid regions contained within Influenza A virus genomes that encode for the hemagglutinin (H5) and neuraminidase (N1) genes. Since the Since the Infleunza A Virus (H5N1) contains an RNA genome, initial amplification reactions are performed using the reverse transcription (RT) primer for 1st strand cDNA synthesis, followed by Polymerase Chain Reaction (PCR) amplification utilizing the forward Amplifluor hairpin primer and reverse primer to produce target-specific fluorescent amplicons. The Amplifluor hairpin primer is labeled with Donor (fluorescein) and Quencher (DABSYL) fluorophores, which produce the high signal to noise ratio observed in the Amplifluor assay. Upon incorporation into new amplicons, the Amplifluor primer is fluorescent (reported in the instrument′s FAM layer), while the fluorescein signal of the unincorporated Amplifluor primer is quenched. When performed on a thermal cycler capable of real-time fluorescence detection, the amplification products of the PCR reaction can be monitored directly in a quantitative fashion, allowing evaluation of template detection in a single, closed-tube system with no additional assay procedure. Where required, template standard curves may be employed to quantitatively determine viral load.
Application
The TM=["Amplifluor"] ID H5N1 Kit is intended for the quantitative or qualitative evaluation of Avian Influenza A H5N1 RNA.
For Research Use Only
For Research Use Only
Components
25X Amplifluor® ID Influenza A, H5 Forward Primer, 100 reactions. Light sensitive. Store at or below -20°C. 25X Amplifluor® ID Influenza A H5 Reverse Primer, 100 reactions. Store at or below -20°C. 25X Amplifluor® ID Influenza A N1 Forward Primer, 100 reactions. Light sensitive, Store at or below -20°C. 25X Amplifluor® ID Influenza A N1 Reverse Primer, 100 reactions. Store at or below -20°C. Positive Control for H5 quantitation, 100 reactions. Store at or below -20°C. Positive Control for N1 quantitation, 100 reactions. Store at or below -20°C.
Storage and Stability
The Amplifluor® ID primers must be stored in the dark at -20°C (or at -80°C for long term storage). Amplifluor® primers are light sensitive. Amplifluor® primers may be divided into convenient aliquots and wrapped in foil to minimize light exposure. Warnings and Precautions: • Handle all specimens and materials coming in contact with them as potentially infectious materials. Decontaminate with 0.05% sodium hypochlorite (1:100 dilution of household bleach). • Do not use reagents beyond expiration date. • Nucleic acids are subject to degradation by nucleases found in the environment and on human surfaces. Clean and cover work surfaces with disposable coverings and wear powder free gloves during all procedures. • Use pipetting techniques that will deliver correct volumes of reagents in all steps, including pre-wetting Aerosol® resistant pipette tips. Repeating pipettors should be primed and voided of all air bubbles. • With nucleic acid amplification, the potential for carry-over contamination is high. Work in a unidirectional flow from workstation to workstation and always use dedicated equipment and Aerosol®-barrier pipette tips.
Legal Information
Aerosol is a registered trademark of Cytec Technology Corp.
Amplifluor is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Certificates of Analysis (COA)
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