P12-17EVALUATION OF THE ROCHE COBAS®HPV ASSAY WITH COLLI-PEETM COLLECTED, UCM PRESERVED URINE.

10. HPV testing
A. Vorsters 1, S. Van Keer 1, S. Biesmans 1, V. Vankerckhoven 2, M. De Koeijer 2, K. Beyers 2, L. Ceelen 3, A. Van Roost 4, P. Van Damme 1.
1University of Antwerp, VAXINFECTIO (Belgium), 2Novosanis (Belgium), 3Anacura (Belgium), 4Roche (Belgium)

Background / Objectives

HPV testing in urine has been proposed for monitoring impact of vaccination, follow-up of treatment and/or reaching women not participating in a cervical cancer screening programme.  The use of Colli-PeeTM (Novosanis, Belgium) and UCM (Urine Collection Medium, UAntwerp, Belgium) has enhanced the analytical detection of HPV DNA in female urine. The Roche Cobas®HPV assay has been validated for clinical sensitive during the ATHENA trial enrolling > 47000 women on cervical material.

The aim of this study was to determine if the Roche Cobas®HPV assay is compatible with Colli-PeeTM collected, UCM preserved urine. 


Methods

Forty-four Colli-PeeTM collected, UCM preserved urine samples were analysed. Thirty-two of these samples originated from a cohort of women participating in a therapeutic HPV vaccination trial.  These samples were collected by the participants at home and were send uncooled by mail to the University of Antwerp. All samples were characterised by an in-house HPV type specific (TS) qPCR method (UAntwerp, Belgium) and or by the Optiplex HPV genotyping kit (Diamex). 


Results

All samples were positive for the Roche beta-globin internal control.

If we look for HPV16 or HPV18 positivity, 20 samples were concordant negative, 14 concordant positive,  and 10 were Diamex positive and Cobas®HPV negative. However, the discrepant results were obtained in samples with low MFI values, and low HPV copy numbers. In addition, the Diamex assay and the in-house PCR are performed on DNA extracts obtained after ultra-filtration of the urine sample, therefore these assay are run with DNA enriched samples compared to the Cobas®HPV.

A correlation between the Ct (cycle threshold) values obtained with Cobas®HPV and the in-house TS qPCR is observed for human DNA and HPV DNA. This further confirms the compatibility of the Roche assay and the Colli-PeeTM collected, UCM preserved urine. 


Conclusion

This pilot study demonstrates that the Roche Cobas®HPV assay is compatible with the Colli-PeeTM collected, UCM preserved urine samples. These results are very encouraging to further investigate possible applications of first void urine in combination with Cobas®HPV assay.


References