TEST:

BD Onclarity™ HPV Assay

Extended HR-HPV genotyping using the BD Onclarity assay demonstrated a high prevalence of HR-HPV in this high-risk population, with multiple genotype infections commonly observed. While non-HPV16/18 genotypes predominated overall, HPV16 was more frequently associated with abnormal cytologic and high-grade histologic findings. Given the limited number of biopsy-confirmed high-grade lesions, these associations should be interpreted as descriptive and hypothesis-generating. Cytology remains central to anal cancer screening, with extended HPV genotyping serving as a complementary adjunct rather than a standalone stratification tool. Larger longitudinal studies with systematic histopathologic follow-up are needed to clarify the clinical significance of extended HPV genotyping in this population.

The results show that using self-collected urine to detect high-risk HPV using the INDICAID™ HPV Urine Test is comparable to using the BD Onclarity™ HPV Assay with clinician-collected vaginal samples. This demonstrates the accuracy of the novel INDICAID™ HPV Urine Test. The potential of urine-based screening as an alternative method is significant, particularly in enhancing participation in cervical cancer screening programs. This approach could be beneficial for subpopulations facing barriers to healthcare access.

Based on ten years of HPV-based cervical cancer screening data, this study demonstrates that genotype-specific HR-HPV information obtained through extended genotyping provides clinically relevant risk stratification when interpreted together with cytology and p16/Ki67 dual-stain results. These findings support an integrated screening approach combining molecular HPV testing, cytology, and immunocytochemical biomarkers to improve risk-based triage in cervical cancer screening.

Extended genotyping with HPV-16/18/33/31/39 achieved 100% sensitivity for CIN3 detection, but a moderate specificity. This strategy aligns with the international recommendation for colposcopy referral in women with a PPV of CIN3 larger than 4.0%.

HPV vaccination is significantly impacting both vaccine and non-vaccine type prevalence, reducing the incidence of vaccine types with concomitant increase of certain non-target types. This apparent dynamic equilibrium has important implications for cervical cancer screening and future incidence of cervical disease.

Dry self-samples from a Copan FLOQSwab taken in clinic are likely to have sufficient DNA quality and accuracy for HPV testing compared with wet self-samples and clinician-collected samples if resuspension takes place up to 2 weeks, with storage at room temperature, and using the BD Onclarity assay. Urine samples using the Colli-pee device are likely to be less sensitive for HPV detected from clinician-collected samples.

Analytical and diagnostic performance of samples collected by the three self-sampling devices resulted in similar quality and HPV detection. Hence the sample quality is not a determinant in the choice of sampling device and focus on other factors such as cost, women's preference or size and weight can take precedence.

Extended genotyping classified 11.5% of 1-year HPV-positive cases as new infections. As 1-year HPV positivity accounts for about 60% of first-level colposcopies, this corresponds to about 7% of colposcopies. Baseline single-channel positivity, age, and p16/ki67-negative results may influence this proportion.

Both clinician- and self-collected samples showed adequate cellularity, with potentially false negative HPV results from low cellular content appearing rare. Observed patterns suggest Ct <26 as optimal and Ct <28 as a minimum for program-level quality assurance with the BD Onclarity™ HPV Assay.

The findings suggest that individuals with HPV16, HPV33/58, and HPV31 infections could be referred immediately to colposcopy. Sending individuals with low-risk genotypes, HPV35/39/68, HPV51, or HPV56/59/66 infections with normal cytology back to routine screening could increase screening specificity.

By fulfilling all guideline requirements for clinical sensitivity, specificity, and reproducibility, Allplex can be considered clinically validated for primary cervical screening using clinician-collected SurePath samples. With this study, Allplex also meets the criteria for a second-generation HPV comparator test.

Extended HPV genotyping may support anal cancer screening strategies by providing a potential standalone tool for both screening and triage. Further studies are needed to confirm these findings in larger cohorts.