학술논문
PCR-RFLP and FTIR-based detection of high-risk human papilloma virus for cervical cancer screening and prevention.
Document Type
Academic Journal
Author
Alves Melo IM; Laboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento, Avenida Shishima Hifumi 2911, Urbanova, São José dos Campos, 12244-000, São Paulo, SP, Brazil.; Pereira Viana MR; Laboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento, Avenida Shishima Hifumi 2911, Urbanova, São José dos Campos, 12244-000, São Paulo, SP, Brazil.; Pupin B; Laboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento, Avenida Shishima Hifumi 2911, Urbanova, São José dos Campos, 12244-000, São Paulo, SP, Brazil.; Bhattacharjee TT; Sir John Walsh Research Institute, University of Otago, 310 Great King St, North Dunedin, Dunedin, 9016, New Zealand.; de Azevedo Canevari R; Laboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento, Avenida Shishima Hifumi 2911, Urbanova, São José dos Campos, 12244-000, São Paulo, SP, Brazil.
Source
Publisher: Elsevier B.V Country of Publication: Netherlands NLM ID: 101660999 Publication Model: eCollection Cited Medium: Internet ISSN: 2405-5808 (Electronic) Linking ISSN: 24055808 NLM ISO Abbreviation: Biochem Biophys Rep Subsets: PubMed not MEDLINE
Subject
Language
English
Abstract
Background: 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 time-consuming. 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.
Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(© 2021 The Authors. Published by Elsevier B.V.)
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
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.
Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(© 2021 The Authors. Published by Elsevier B.V.)