artus HSV-1/2 Quant RG PCR Kit CE

• Compliance with EU IVD Directive 98/79/EC
• Detection and quantification of HSV-1 and HSV-2 DNA
• High reliability using the Internal Control
• Highly sensitive and specific
• Covers a broad linear range

The specific performance characteristics of the artus HSV-1/2 Quant RG PCR Kit were determined using HSV-1-specific DNA (ATCC number: VR-1493) and HSV-2-specific DNA (ATCC number: VR-540) of known concentrations.

Analytical sensitivity

The analytical sensitivity of the artus HSV-1/2 Quant RG PCR Kit, determined by probit analysis, for detection of HSV-1-specific DNA is 0.33 copies/µl eluate (95% confidence interval [CI]: 0.16–1.3 copies/µl) and the analytical sensitivity for detection of HSV-2-specific DNA is 1.2 copies/µl eluate (95% CI: 0.7–3.5 copies/µl).

Analytical specificity

The analytical specificity of the artus HSV-1/2 Quant RG PCR Kit is ensured by careful selection of the oligonucleotides (primers and probes). The oligonucleotides are checked by sequence comparison analysis against publically available sequences to ensure that all relevant HSV genotypes are detected. In addition, the specificity of the artus HSV-1/2 Quant RG PCR Kit was evaluated by testing a panel of genomic DNA/RNA extracted from other herpesviruses and pathogens that are relevant to immunocompromised patients. No cross-reactivity was shown.

Linear range

The linear range of the artus HSV-1/2 Quant RG PCR Kit was evaluated by analyzing a logarithmic dilution series of HSV-1- and HSV-2-specific DNA using concentrations ranging from 10⁸ copies/µl to 10 copies/µl (HSV-1) and 10⁷ to 10 copies/µl (HSV-2). At least six replicates per dilution were analyzed. The linear range of the artus HSV-1/2 Quant RG PCR Kit extends over an interval of at least seven orders of magnitude for HSV-1- and over an interval of at least six orders of magnitude for HSV-2-specific DNA.

Sample DNA is first extracted in a separate process e.g., using the QIAamp DNA Mini Kit (recommended extraction method). This is followed by detection of HSV-1- and HSV-2-specific DNA using the Rotor-Gene Q.

The HSV-1/2 RG Master A and HSV-1/2 RG Master B contain reagents and enzymes for the specific amplification of target regions within the HSV-1 and HSV-2 genomes and for the direct detection of the specific amplicon in fluorescence channels Cycling Green and Cycling Red of Rotor-Gene Q instruments.

In addition, the artus HSV-1/2 RG PCR Kit contains a heterologous amplification system (Internal Control) to identify potential failures during the assay process. The Internal Control is measured in the fluorescence channel Cycling Yellow of Rotor-Gene Q instruments.

Probes specific for HSV-1 DNA are labeled with the fluorophore FAM, while probes specific for HSV-2 DNA are labeled with a fluorophore that displays the same characteristics as Cy5. The probe specific for the Internal Control is labeled with the fluorophore JOE. The use of probes labeled with spectrally distinguishable fluorophores enables simultaneous detection and quantification of HSV-1- and HSV-2-specific DNA, as well as detection of the Internal Control in the corresponding channels of the Rotor-Gene Q instrument.

The artus HSV-1/2 Quant RG PCR Kit is for in vitro diagnostic use for detection and quantification of Herpes Simplex Virus 1 (HSV-1) and Herpes Simplex Virus 2 (HSV-2) specific DNA.