artus HAdV RG PCR Kit CE

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

As there is no international standard available for adenovirus, quantitative performance evaluation of the artus HAdV RG PCR Kit was carried out using genomic DNA from a characterized HAdV-3 isolate (species B).

Analytical sensitivity

The analytical sensitivity of the artus HAdV RG PCR Kit, determined by probit analysis, for detection of HAdV-specific DNA is 1.07 copies/µl (95% confidence interval [CI]: 0.58–2.99 copies/μl).

Analytical specificity

The analytical specificity of the artus HAdV 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 adenovirus genotypes are detected. In addition, the specificity of the artus HAdV RG PCR Kit was evaluated by testing a panel of genomic DNA/RNA extracted from other pathogens causing similar symptoms as adenovirus infections and by testing human genomic DNA. No cross-reactivity was shown.

Linear range

The linear range of the artus HAdV RG PCR Kit was evaluated by analyzing a logarithmic dilution series of quantified genomic HAdV-2 DNA (species C) using concentrations ranging from 1 x 10⁹ to 0.1 copies/µl. Six replicates per dilution were analyzed. The linear range of the artus HAdV RG PCR Kit extends over an interval of at least eight orders of magnitude for HAdV-specific DNA.

Diagnostic evaluation

The diagnostic sensitivity and specificity of theartus HAdV RG PCR Kit are regularly evaluated by analyzing reference and diagnostic samples previously analyzed with reference methods (i.e., in-house PCR, DFA, shell vial culture, electron microscopy, Luminex technology). So far, 223 specimens derived from smears, nasopharyngeal aspirates, bronchial secretions, stool samples, urine samples, plasma or eye smears collected in different laboratories were tested for determining the diagnostic sensitivity and specificity of the artus HAdV RG PCR Kit. Out of these 223 specimens, 50 were predicated to be HAdV positive and 173 were predicated to be HAdV negative by reference methods.
Four samples tested positive for HAdV (C_T values 35.2, 36.8, 40.0 and 37.9) with the artus HAdV RG PCR Kit that previously tested negative with an in-house PCR test. All 50 specimens predicted to contain HAdV DNA were confirmed as HAdV positive by analysis with the artus HAdV RG PCR Kit.

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 HAdV-specific DNA using the Rotor-Gene Q.

The HAdV RG Master A and HAdV RG Master B contain reagents and enzymes for the specific amplification of target regions within the HAdV genome and for the direct detection of the specific amplicon in fluorescence channel Cycling Green of Rotor-Gene Q instruments.

In addition, the artus HAdV 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 HAdV DNA are labeled with the fluorophore FAM. 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 HAdV DNA as well as detection of the Internal Control in the corresponding channels of the Rotor-Gene Q instrument.

The artus HAdV RG PCR Kit is for in vitro diagnostic use for detection and quantification of human adenovirus (HAdV) specific DNA.