artus HAdV RG PCR Kit CE
For quantitative detection of human adenovirus (HAdV) DNA using real-time PCR
The artus HAdV RG PCR Kit is a real-time PCR test for the detection and quantification of human adenovirus (HAdV) specific DNA using real-time PCR on the Rotor-Gene Q.
The artus HAdV RG PCR Kit CE is intended for in vitro diagnostic use in Europe.
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.
The artus HAdV RG PCR Kit constitutes a ready-to-use system for the detection and quantification of HAdV-specific DNA using real-time PCR on Rotor-Gene Q instruments. The assay includes a heterologous amplification system (Internal Control) to identify possible PCR inhibition and to confirm the integrity of the kit reagents.
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.
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