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).
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).
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.
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.
The diagnostic sensitivity and specificity of the artus 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 (CT 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.
| ||Negative || Positive |
| Negative || 169 || 4* |
| Positive || 0 || 50 |