P08-14Comparison of Seegene Anyplex II HPV 28 detection and Abbott Realtime High Risk HPV test on NOVAprep liquid-based cytology media.

08. HPV testing
S. Hantz 1, C. Deback 2, C. Gaudy-Graffin 3, J. Darreye 1, G. Darreye 4, S. Alain 1.
1Laboratory of Bacteriology-Virology-Hygiene, CHU Limoges, Limoges (France), 2Laboratory of Virology, CHU Paul Brousse, Villejuif (France), 3Laboratory of Virology, CHU Tours, Tours (France), 4Laboratory of Pathology, Limoges (France)

Background / Objectives

High Risk (HR) HPV DNA testing is a highly sensitive method to screen women at risk of CIN2+ lesions. But many assays and various media for cervical sample collection are available. So, we aimed to compare the Seegene AnyplexTM II HPV 28 Detection and the Abbott RealTime High Risk HPV test on cervical samples collected on NOVAprep® liquid based cytology media (Novacyt).


Methods

Samples were collected on NOVAprep® media from July 2016 to February 2017. Cytology was performed with NOVAprep® liquid-based cytology platform for cervical cancer screening. Samples with ASCUS cytology were routinely tested with the Abbott assay for HPV testing (14 HR HPV detected:16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68). Among them, 162 samples were randomly selected for testing with the Seegene assay after extraction on Easymag (Biomérieux) following manufacturer’s recommendations. Anyplex™ II HPV28 Detection simultaneously detects 19 HR HPV (16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 69, 73, 82) and 9 low-risk HPV (6,11, 40, 42, 43, 44, 54, 61, 70), with semi-quantitative analysis (+, ++ or +++). Discordant results were confirmed by testing again both assays at the same time and then will be sequenced by NGS


Results

All the 162 samples have interpretable results as internal control was valid in both assays. Global HR HPV prevalence was more elevated with Seegene assay (49.38%) than Abbott assay (37.65%). Regarding the 14 HR genotypes detectable by both assays, overall agreement between both assays was very good (93%; kappa 0.86). All discordant results were confirmed in second runs (Seegene and Abbott), showing excellent reproducibility of each assay. Among the 10 Abbott HR HPV negative samples detected as HR HPV by Seegene, analysis of Abbott amplification curves showed that HR HPV (other than HPV16 or HPV18) were detected in 2 samples after 32 Ct (detection threshold determined by Abbott). For both samples, semi-quantification from Seegene assay found a small amount of HR HPV (only one +). Among these discordant results, HPV 68 is the less efficiently detected HPV by Abbott assay (6/10). Small amounts of HR HPV (one +) were also found in three of the four other discordant results. Only one HR positive with Abbott assay was not detected by Seegene assay. Complete analysis of discordant results by NGS is ongoing.


Conclusion

NOVAprep® medium previously validated with Hybrid Capture assay has demonstrated again strong performances in terms of detection of HR HPV and stability of the patient samples with Abbott RealTime High Risk HPV test and Seegene AnyplexTM II HPV 28 Detection assays.


References

1: Guenat D, Launay S, Riethmuller D, Mougin C, Prétet JL. Validation of Novaprep(®) HQ+ liquid-based cytology medium for high-risk human papillomavirus detection by hc2. Infect Agent Cancer. 2016;11:41.

2: Khiri H, Camus C, Portugal M, Pénaranda G, Boyer S, Halfon P. [Cytological and virological medium performance and stability assessment using the Cobas 4800 HPV  test (Roche Diagnostics) used in France]. Ann Biol Clin (Paris). 2014;72:213-23

3: Prétet JL, Vidal C, Le Bail Carval K, Ramanah R, Carcopino X, Cartier I, Labouyrie E, Kantelip B, Coumes-Marquet S, Riethmuller D, Mougin C. Novaprep(®) Vial Test is a suitable liquid-based cytology medium for high risk human papillomavirus testing by Hybrid Capture 2. J Clin Virol. 2010;49:286-9.