Multiplex Probe Amplification (MPA) is a patented real-time PCR-based technology allowing detection and genotyping of up to 20 different targets in a single closed-tube reaction, thus significantly increases throughput capability. Papilloplex™ HR HPV is a CE IVD marked product for qualitative detection and differentiation of all 14 high-risk HPV types in a single analysis.
In the present study, we carried out a comparative analysis of the performance of Papilloplex™ HR HPV test with other well established assays on clinical samples. Analytical specificity of the assay was also interrogated using the WHO HPV LabNet proficiency panel.
The Papilloplex™ HR HPV test was applied to 500 disease enriched cervical liquid based cytology samples obtained from the Scottish HPV Archive, Edinburgh, with known concurrent pathology results. Samples were also tested using the Abbott RealTime High Risk HPV assay, the Qiagen digene HC2 HPV DNA Test, the Diamex Optiplex HPV Genotyping kit and Roche Linear Array® HPV Genotyping Test. Concordance between the comparator assays and Papilloplex™ HR HPV was performed using both binomial and McNemar’s test.
The Papilloplex™ HR HPV was able to detect and genotype high risk types 16, 18, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68a and 68b in a single reaction. The limit of detection for the assay was up to 5 genome copy numbers for HPV 16 and 18 in WHO LabNet samples with 100% accuracy for genotyping. No significant difference in the qualitative detection of high risk HPV was observed between the Papilloplex™ HR HPV and the four assays described above. Type-specific concordance was also high.
In conclusion, this study shows that the Papilloplex™ HR HPV is efficient in combined screening and genotyping of HPV DNA. The current commercially available probe-based methods are limited to detection of only one target sequence per fluorescence channel. The MPA technology overcomes this limitation, allowing 14 targets to be detected and quantified in a single closed-tube reaction.
These data indicate that the analytical performance of Papilloplex™ HR HPV is comparable to established HPV assays at the level of generic high risk HPV detection and at the type-specific level. The assay shows potential promise from both disease management and epidemiological perspectives.