artus HHV-6 RG PCR Kit CE

• Compliance with EU IVD Directive 98/79/EC
• Detection and differentiation of HHV-6A and HHV-6B DNA
• High reliability using the Internal Control
• Highly sensitive and specific
• Covers a broad linear range

The specific performance characteristics of the artus HHV-6 RG PCR Kit were determined using HHV-6A-specific DNA (strain GS) and HHV-6B-specific DNA (strain Z-29) of known concentrations.

Analytical sensitivity

The analytical sensitivity of the artus HHV-6 RG PCR Kit, determined by probit analysis, for detection of HHV-6A-specific DNA is 1.46 copies/µl eluate [95% confidence interval (CI): 0.72–4.57 copies/µl] and the analytical sensitivity for detection of HHV-6B-specific DNA is 2.58 copies/µl eluate (95% CI: 1.44–6.30 copies/µl).

Analytical specificity

The analytical specificity of the artus HHV-6 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 HHV-6 genotypes are detected. In addition, the specificity of the artus HHV-6 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 HHV-6 RG PCR Kit was evaluated by analyzing a logarithmic dilution series of HHV-6A- and HHV-6B-specific DNA using concentrations ranging from 10⁸ copies/µl to 1 copy/µl. At least six replicates per dilution were analyzed. The linear range of the artus HHV-6 RG PCR Kit extends over an interval of at least seven orders of magnitude for HHV-6A- and HHV-6B-specific DNA.

Diagnostic evaluation

The diagnostic sensitivity and specificity of the artus HHV-6 RG PCR Kit are regularly evaluated by analyzing reference and diagnostic samples previously analyzed with reference methods.

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 HHV-6A- and HHV-6B-specific DNA using the Rotor-Gene Q.

The HHV-6 RG Master A and HHV-6 RG Master B contain reagents and enzymes for the specific amplification of target regions within the HHV-6A and HHV-6B 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 HHV-6 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 HHV-6A DNA are labeled with the fluorophore FAM, while probes specific for HHV-6B 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 HHV-6A- and HHV-6B-specific DNA as well as detection of the Internal Control in the corresponding channels of the Rotor-Gene Q instrument.

The artus HHV-6 RG PCR Kit is for in vitro diagnostic use for detection, differentiation and quantification of Human Herpesvirus 6A (HHV-6A) and Human Herpesvirus 6B (HHV-6B) specific DNA.