P27-03COMPARISON OF ANYPLEX II HPV 28 AND SPF10/DEIA/LiPA25 IN FFPE OROPHARYNGEAL CANCER SAMPLES

Human Papillomavirus (HPV) infection has emerged as a major etiological factor with a prognostic significance in oropharyngeal squamous cell carcinomas (OSCC). The identification of HPV-related tumors in the clinical setting is still mainly based on the p16^INK4a overexpression. However, a recently published meta-analysis (Prigge ES, IJC 2017) shows that the combination between p16 overexpression and HPV DNA PCR testing increases the specificity to distinguish HPV-related OSCC. Several techniques focused on HPV DNA detection and genotyping, which have been developed and used for prognosis purposes, could also be used in the near future for treatment decision-making. Here, we compare the results obtained analyzing formalin-fixed paraffin-embedded (FFPE) OSCC tissue blocks using the Anyplex II HPV 28 test and two different extraction methods with the results previously obtained using the SPF10/DEIA/LiPA25 combination for HPV DNA detection and genotyping.

FFPE samples were selected from an international collection including 1090 OSCCs subjected to histopathological evaluation, HPV-DNA, HPV E6*I mRNA and p16^INK4a detection (Castellsagué/Alemany, JNCI, 2016). A subset of 95 FFPE samples were randomly selected from strata specified (40 HPV DNA negative; 40 HPV DNA positive + mRNA or p16 positive; 12 HPV DNA negative+p16 positive; 3 HPV DNA positive + mRNA/p16 negative). DNA extraction for HPV DNA detection (SPF10/DEIA/LiPA25) was performed using a Proteinase K digestion method as previously described. In the present study, samples have been retested using Anyplex II HPV 28 test and two different DNA extraction methods. Proteinase K digestion method and DNA extraction using the MagCore automatic station were compared. We calculate the agreement between the SPF10/DEIA/LiPA25 and Anyplex II HPV 28 test.

Preliminary data obtained after the analysis of 32 samples show 100% agreement between SPF10/DEIA/LiPA25 and Anyplex II HPV 28 tests.  Anyplex II HPV 28 test was positive for 58% and negative for 42% of samples. All HPV positive samples showed the HPV16 type. A complete agreement in HPV detection and genotyping was obtained using Proteinase K digestion method and the MagCore protocol.

Preliminary data have shown a high agreement between SPF10/DEIA/LiPA25 and Anyplex II HPV 28 tests for HPV detection and typing in FFPE oropharyngeal samples. This is an on-going study and complete results will be presented in EUROGIN 2017 congress.

^{-Prigge ES, Arbyn M, von Knebel Doeberitz M, Reuschenbach M. Diagnostic accuracy of p16(INK4a) immunohistochemistry in oropharyngeal squamous cell carcinomas: A systematic review and meta-analysis. Int J Cancer. 2017 Mar 1;140(5):1186-1198.}

^{-Castellsagué X, Alemany L, Quer M, Halec G, Quirós B, Tous S, Clavero O, Alòs L, Biegner T, Szafarowski T, Alejo M, Holzinger D, Cadena E, Claros E, Hall G, Laco J, Poljak M, Benevolo M, Kasamatsu E, Mehanna H, Ndiaye C, Guimerà N, Lloveras B, León X, Ruiz-Cabezas JC, Alvarado-Cabrero I, Kang CS, Oh JK, Garcia-Rojo M, Iljazovic E, Ajayi OF, Duarte F, Nessa A, Tinoco L, Duran-Padilla MA, Pirog EC, Viarheichyk H, Morales H, Costes V, Félix A, Germar MJ, Mena M, Ruacan A, Jain A, Mehrotra R, Goodman MT, Lombardi LE, Ferrera A, Malami S, Albanesi EI, Dabed P, Molina C, López-Revilla R, Mandys V, González ME, Velasco J, Bravo IG, Quint W, Pawlita M, Muñoz N, de Sanjosé S, Xavier Bosch F; ICO International HPV in Head and Neck Cancer Study Group. HPV Involvement in Head and Neck Cancers: Comprehensive Assessment of Biomarkers in 3680 Patients. J Natl Cancer Inst. 2016 Jan 28;108(6).}