Human Papillomavirus DNA Detection by Droplet Digital PCR in Formalin-Fixed Paraffin-Embedded Tumor Tissue from Oropharyngeal Squamous Cell Carcinoma Patients

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Schiavetto, Camila Marques ^[1] ; de Abreu, Priscila Marinho ^[2] ; von Zeidler, Sandra Ventorin ^[2] ; de Jesus, Lais Machado ^[1] ; Carvalho, Raiany Santos ^[3] ; Cirino, Maria Thereza ^[1] ; Carloni, Adriana Cruvinel ^[1] ; Oliveira, Cristina ^[1] ; Scapulatempo-Neto, Cristovam ^{[1, 4]} ; de Almeida, Gisele Caravina ^{[5, 4]} ; de Menezes, Nei Soares ^[5] ; Carvalho, Andre Lopes ^[1] ; Reis, Rui Manuel ^{[6, 7, 1]} ; de Carvalho, Ana Carolina ^[1]

Afiliação do(s) autor(es)

^[1] Barretos Canc Hosp, Mol Oncol Res Ctr, Rua Antenor Duarte Vilela 1331, BR-14784400 Barretos, SP - Brazil

^[2] Univ Fed Espirito Santo, Programa Posgrad Biotecnol, Ctr Ciencias Saude, Vitoria, ES - Brazil

^[3] Barretos Canc Hosp, Res Support Ctr, Sao Paulo - Brazil

^[4] Diagnost Amer DASA, Pathol & Mol Diagnost Serv, Sao Paulo - Brazil

^[5] Barretos Canc Hosp, Dept Pathol, Sao Paulo - Brazil

^[6] ICVS 3Bs PT Govt Associate Lab, Braga - Portugal

^[7] Univ Minho, Sch Med, Life & Hlth Sci Res Inst ICVS, Braga - Portugal

• Tipo de documento: Artigo Científico
• Fonte: MOLECULAR DIAGNOSIS & THERAPY; v. 25, n. 1 NOV 2020.
• Citações Web of Science: 0

Resumo

Introduction High-risk human papillomavirus infection impacts staging and prognosis of oropharyngeal squamous cell carcinomas (OPSCCs). Determination of HPV status in tumor tissue by p16-immunohistochemistry (p16-IHC) can be challenging; therefore, complementary methodologies could be useful in a clinical setting. Objective To test for accuracy and clinical relevance of HPV-DNA detection in formalin-fixed and paraffin-embedded (FFPE) tumor samples by droplet digital PCR (ddPCR). Materials and Methods Fifty OPSCCs were tested for p16-IHC status followed by HPV-16/18 DNA detection/quantification in FFPE-recovered DNA using ddPCR. Accuracy for HPV status determination and association with patient information were also evaluated. Results 32.0% (16/50) of the cases were p16-IHC positive (p16 +), 42.0% (21/50) had detectable levels of HPV-16 DNA, and none were positive for HPV-18 DNA. A higher median viral load of HPV-16 DNA was observed in p16 + cases (p < 0.0001). Concordance between p16-IHC and HPV-16 DNA ranged from 78.0 to 86.0% and accuracy rates were between 78.0 and 86.0%. P16-IHC and HPV-16 DNA detection was associated with gender, smoking status, and tumor subsite, while only HPV-16 DNA was associated with cT stage. The combination of HPV positivity by p16-IHC and ddPCR showed higher overall survival rates in comparison with p16 + /HPV-DNA- and p16 - /HPV-DNA- results. Conclusions Type-specific HPV-DNA detection by ddPCR is highly specific but moderately sensitive for the determination of HPV status and showed clinical relevance, mainly when associated with p16-IHC status. Results highlight the importance of performing HPV-DNA testing in combination with p16-IHC for proper identification of HPV-associated OPSCC and to improve clinical management of OPSCC patients. (AU)

• Processo FAPESP: 15/01286-0 - Caracterização de alterações genéticas e epigenéticas em genes específicos em tumores HPV-positivos e HPV-negativos de pacientes com carcinoma epidermóide de cabeça e pescoço
• Beneficiário: André Lopes Carvalho
• Modalidade de apoio: Auxílio à Pesquisa - Regular