Herpes simplex virus type 1 (HSV1), Herpes simplex virus type 2 (HSV2) and Varicella zoster virus (VZV) PCR Kit CE/IVD
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RealCycler HHVA-
Catalog Number: RealCycler HHVA-
1 Intended use:
RealCycler HHVA is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of DNA from Herpesvirus type 1, Herpesvirus type 2 and Varicella-Zoster Virus simultaneously in clinical samples.
HSV1, HSV2 and VZV pathogens could be quantified if this product is used in combination with the corresponding calibrator (references RealCycler HV1Q, HV2Q orVZVQ respectively).
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 HSVT amplification, Herpesvirus type 1 is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 fluorophore and Herpesvirus type 2 is detected in the corresponding channel of TxR fluorophore.
In the VARI amplification, Varicella-Zoster virus is detected in the corresponding channel of FAM fluorophore and internal control is detected in the corresponding channel of Alx532 fluorophore.
3 Technical specifications
Sensitivity:
- Herpesvirus type 1: 10 copies/μL ,
- Herpesvirus type 2: copy/μL and
- Varicella-Zoster Virus: 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:
- Herpes virus type 11: US6 gene,
- Herpesvirus type 2: DNA polymerase gene and
- Varicella-Zoster Virus: Single-stranded DNA-binding protein gene (ORF 29)).
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler HHVA includes the HSVT AmpliMix and VARI AmpliMix, a HSVT DNA Positive Control, which contains a mixture of Herpes virus type 1 and Herpes virus type 2 DNA and a VZV DNA Positive Control.
All reagents are ready to use without adding or rebuilding any component.
RealCycler HHVA is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of DNA from Herpesvirus type 1, Herpesvirus type 2 and Varicella-Zoster Virus simultaneously in clinical samples.
HSV1, HSV2 and VZV pathogens could be quantified if this product is used in combination with the corresponding calibrator (references RealCycler HV1Q, HV2Q orVZVQ respectively).
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 HSVT amplification, Herpesvirus type 1 is detected in the corresponding channel of FAM fluorophore, CHIC is detected in the corresponding channel of Alx532 fluorophore and Herpesvirus type 2 is detected in the corresponding channel of TxR fluorophore.
In the VARI amplification, Varicella-Zoster virus is detected in the corresponding channel of FAM fluorophore and internal control is detected in the corresponding channel of Alx532 fluorophore.
3 Technical specifications
Sensitivity:
- Herpesvirus type 1: 10 copies/μL ,
- Herpesvirus type 2: copy/μL and
- Varicella-Zoster Virus: 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:
- Herpes virus type 11: US6 gene,
- Herpesvirus type 2: DNA polymerase gene and
- Varicella-Zoster Virus: Single-stranded DNA-binding protein gene (ORF 29)).
Specificity validation has been performed according to experimental assays and BLAST analysis (www.ncbi.nlm.nih.gov/blast).
4 Contents
RealCycler HHVA includes the HSVT AmpliMix and VARI AmpliMix, a HSVT DNA Positive Control, which contains a mixture of Herpes virus type 1 and Herpes virus type 2 DNA and a VZV DNA Positive Control.
All reagents are ready to use without adding or rebuilding any component.
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