FC 05-09HPV DNA GENOTYPE AGREEMENT AND CLINICAL PERFORMANCE IN FIRST-VOID URINE AND CERVICAL SAMPLES IN A REFERRAL POPULATION IN BELGIUM

08. HPV testing
S. Van Keer 1, J. Pattyn 1, S. Biesmans 1, X. Van Ostade 2, M. Ieven 3, W. Tjalma 4, P. Van Damme 1, A. Vorsters 1.
1Centre for the Evaluation of Vaccination (CEV); Vaccine & Infectious Disease Institute (VAXINFECTIO); Faculty of Medicine and Health Sciences; University of Antwerp (Belgium), 2Proteomics; Proteinscience, Proteomics & Epigenetic Signaling (PPES); Faculty of Pharmaceutical, Biomedical and Veterinary Sciences; University of Antwerp (Belgium), 3Laboratory of Medical Microbiology (LMM); Vaccine & Infectious Disease Institute (VAXINFECTIO); Faculty of Medicine and Health Sciences; University of Antwerp (Belgium), 4Multidisciplinary Breast Clinic, Unit Gynaecologic Oncology; Department of Obstetrics and Gynaecology; Molecular Imaging, Pathology, Radiotherapy, Oncology (MIPRO); Faculty of Medicine and Health Sciences; Antwerp University Hospital (UZA) - University of Antwerp (Belgium)

Background / Objectives

This study reports on the performance and acceptability of optimized HPV detection and genotyping in self-collected first-void urine (FvU) versus cervical (Cx) samples for the detection of high-risk (hr)HPV DNA and cervical precancerous lesions in a referral population in Belgium.


Methods

Women between 25-64 years old (median age: 36.00 ± 10.08 year) referred for colposcopy at the Antwerp University Hospital (UZA) collected a FvU sample (Colli-PeeTM, Novosanis) (NCT02714127) prior to their visit with the gynecologist. HPV DNA genotyping was performed on paired FvU (after in-house DNA extraction (1)) and Cervex-Brush® (Rovers Medical Devices) collected Cx samples in PreservCyt® (Hologic) with the Riatol qPCR HPV genotyping assay (Belgium) (2). Histology on biopsies (when indicated) was investigated at the pathology laboratory (UZA). Data regarding acceptability of different sampling methods were gathered through questionnaires. Statistics was performed using IBM SPSS24.


Results

HrHPV DNA was detected in 69.09 (n=76/110) and 66.36% (n=73/110) of FvU and Cx samples respectively, with HPV16 the most prevalent genotype (n=23/110 versus n=21/110). A good agreement for hrHPV DNA in FvU and Cx samples of 86.36% (Cohen’s kappa: 0.688; 95% CI: 0.543-0.833) was found. On individual genotype level, excellent agreement for HPV16 (96.36%; Cohen’s kappa: 0.886; 95% CI: 0.776-0.996) and HPV18 (99.09%; Cohen’s kappa: 0.918; 95% CI: 0.759-1.077) was obtained. Moreover, significant positive correlations of HPV16 and 18 DNA copies per µl DNA extract were found between both sample types (Spearman rho HPV16: 0.570, p-value: 0.009; and HPV18: 0.829, p-value: 0.042). For 33 out of 110 samples with histological reference, a relative sensitivity for CIN2+/CIN3+ and specificity for <CIN2 of HPV16/18 detection in FvU versus Cx samples of 1.00 (both CIN2+ and CIN3+) and 0.93 (<CIN2) was acquired.

Preference of using a female urination device upon FvU collection compared to use of a standard urine cup, a pap smear taken by a clinician, or use of a vaginal self-sampling device was respectively 91.75 (n=89/97), 77.36 (n=82/106), and 91.67% (n=11/12).


Conclusion

Good to excellent agreement between FvU and Cx samples on hrHPV DNA and HPV16/18 genotype level was obtained, with significant positive correlations in HPV16/18 copies per µl DNA between both samples. These results furthermore demonstrate that FvU self-sampling is highly preferred among 25-64 year old women referred to colposcopy in Belgium.


References

1. Vorsters A, Van den Bergh J, Micalessi I, Biesmans S, Bogers J, Hens A, et al. Optimization of HPV DNA detection in urine by improving collection, storage, and extraction. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology. 2014;33(11):2005-14.
2. Micalessi IM, Boulet GAV, Bogers JJ, Benoy IH, Depuydt CE. High-throughput detection, genotyping and quantification of the human papillomavirus using real-time PCR. Clin Chem Lab Med. 2012;50(4):655-61.