PCR-RFLP and FTIR-based detection of high-risk human papilloma virus for cervical cancer screening and prevention

dc.contributor.authorMelo, Igor Martins Alves
dc.contributor.authorViana, Magda Rogéria Pereira
dc.contributor.authorPupin, Breno
dc.contributor.authorBhattacharjee, Tanmoy Tapobrata
dc.contributor.authorCanevari, Renata de Azevedo
dc.date.accessioned2025-04-22T13:26:39Z
dc.date.available2025-04-22T13:26:39Z
dc.date.issued22021-03-25
dc.description.abstractBackground: Approximately 70% of cervical carcinoma cases show the presence of high-risk Human Papilloma Virus (HPV), especially HPV-16 and HPV-18, and can be used to stratify high risk patients from low risk and healthy. Currently, molecular biology techniques such as polymerase chain reaction (PCR) are used to identify the presence of virus in patient samples. While the methodology is highly sensitive, it is labor intensive and timeconsuming. Alternative techniques, such as vibrational spectroscopy, has been suggested as a possible rapid alternative. Therefore, in this study, we evaluate the efficiency of cervical fluid Fourier Transform Infrared spectroscopy (FTIR) in patient risk stratification informed by PCR. Methods: Cervical fluid samples (n = 91) were obtained from patients who have undergone routine Papanicolaou (Pap) test. Viral genome was identified and classified as high/low-risk by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP). FTIR spectra were acquired from samples identified by PCR-RFLP as No-HPV (n = 10), high-risk HPV (n = 7), and low-risk HPV (n = 7). Results: Of the 91 samples, was detected the viral genome by PCR in 36 samples. Of these 36 samples, nine samples were identified to contain high-risk HPV (HR-HPV) and nine samples were found to have low-risk HPV (LR-HPV). The FTIR spectra acquired from No-HPV, LR-HPV, and HR-HPV showed differences in 1069, 1437, 1555, 1647, 2840, 2919, and 3287 cm-1 bands. Principal Component Analysis (PCA) showed distinct clusters for No-HPV and HR-HPV and No-HPV and LR-HPV, but there was significant overlap in the clusters of HR-HPV and LR-HPV. PCA-Linear Discriminant Analysis (PC-LDA) after Leave One Out Cross Validation (LOOCV) classified No-HPV from HR-HPV and No-HPV from LR-HPV with 100% efficiency in the 1400-1800 cm-1 spectral range. LOOCV classifications for LR-HPV and HR-HPV from each other were 71 and 75%, respectively, in the 2800-3400 cm-1 spectral range. Conclusions: The results highlight the high sensitivity of PCR-RFLP in HPV identification and show that FTIR can classify samples identified as healthy, low, and high-risk samples by PCR-RFLP. General significance: We show the possibility of using FTIR for initial cervical cancer risk stratification followed by detailed PCR-RFLP investigations for suspect cases.
dc.description.physical6 p.
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.uriFAPESP: Project 2013/14464–9; CNPq: Project 800985/2018–4.
dc.format.mimetypepdf
dc.identifier.affiliationLaboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento
dc.identifier.affiliationLaboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento
dc.identifier.affiliationLaboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento
dc.identifier.affiliationSir John Walsh Research Institute, University of Otago
dc.identifier.affiliationLaboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento
dc.identifier.bibliographicCitationALVES MELO, I. M. et al. PCR-RFLP and FTIR-based detection of high-risk human papilloma virus for cervical cancer screening and prevention. Biochemistry and Biophysics Reports, [s. l.], v. 26, p. 100993, 2021. Disponível em: https://linkinghub.elsevier.com/retrieve/pii/S240558082100087X. A
dc.identifier.doihttps://doi.org/10.1016/j.bbrep.2021.100993
dc.identifier.urihttps://repositorio.univap.br/handle/123456789/861
dc.language.isoen
dc.publisherElsevier
dc.rights.holderIgor Martins Alves Melo
dc.rights.holderMagda Rogéria Pereira Viana
dc.rights.holderBreno Pupin
dc.rights.holderTanmoy Tapobrata Bhattacharjee
dc.rights.holderRenata de Azevedo Canevari
dc.subject.keywordVírus do papiloma humano (HPV)
dc.subject.keywordCarcinoma cervical
dc.subject.keywordEspectroscopia de infravermelho
dc.subject.keywordFTIR
dc.titlePCR-RFLP and FTIR-based detection of high-risk human papilloma virus for cervical cancer screening and prevention
dc.typeArtigos de Periódicos

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