Comparison of four different human papillomavirus genotyping methods in cervical samples: Addressing method-specific advantages and limitations
Juliana D. Siqueira,
Brunna M. Alves,
Adriana B.C. Castelo Branco,
Kristiane C.D. Duque,
Maria Teresa Bustamante-Teixeira,
Esmeralda A. Soares,
José Eduardo Levi,
Gulnar Azevedo e Silva,
Marcelo A. Soares
Affiliations
Juliana D. Siqueira
Programa de Oncovirologia, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brazil
Brunna M. Alves
Programa de Oncovirologia, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brazil
Adriana B.C. Castelo Branco
Programa de Oncovirologia, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brazil
Kristiane C.D. Duque
Diretoria de Ensino, Pesquisa e Extensão, Instituto Federal de Santa Catarina, Joinville, SC, Brazil
Maria Teresa Bustamante-Teixeira
Núcleo de Assessoria, Treinamento e Estudos em Saúde, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
Esmeralda A. Soares
Programa de Oncovirologia, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brazil
José Eduardo Levi
Instituto de Medicina Tropical de São Paulo Medical School, Universidade de São Paulo, São Paulo, Brazil; Pesquisa e Desenvolvimento, Dasa Laboratories, Barueri, SP, Brazil
Gulnar Azevedo e Silva
Departamento de Epidemiologia, Instituto de Medicina Social, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
Marcelo A. Soares
Programa de Oncovirologia, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brazil; Departamento de Genética, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil; Corresponding author. Instituto Nacional de Câncer Rua André Cavalcanti, 37 – 4o andar 20231-050, Rio de Janeiro, Brazil.
Since human papillomavirus (HPV) is recognized as the causative agent of cervical cancer and associated with anogenital non-cervical and oropharyngeal cancers, the characterization of the HPV types circulating in different geographic regions is an important tool in screening and prevention. In this context, this study compared four methodologies for HPV detection and genotyping: real-time PCR (Cobas® HPV test), nested PCR followed by conventional Sanger sequencing, reverse hybridization (High + Low PapillomaStrip® kit) and next-generation sequencing (NGS) at an Illumina HiSeq2500 platform. Cervical samples from patients followed at the Family Health Strategy from Juiz de Fora, Minas Gerais, Brazil, were collected and subjected to the real-time PCR. Of those, 114 were included in this study according to the results obtained with the real-time PCR, considered herein as the gold standard method. For the 110 samples tested by at least one methodology in addition to real-time PCR, NGS showed the lowest concordance rates of HPV and high-risk HPV identification compared to the other three methods (67–75 %). Real-time PCR and Sanger sequencing showed the highest rates of concordance (97–100 %). All methods differed in their sensitivity and specificity. HPV genotyping contributes to individual risk stratification, therapeutic decisions, epidemiological studies and vaccine development, supporting approaches in prevention, healthcare and management of HPV infection.
