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      A Low-Cost HPV Immunochromatographic Assay to Detect High-Grade Cervical Intraepithelial Neoplasia

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          Abstract

          Objective

          To evaluate the reproducibility and accuracy of the HPV16/18-E6 test.

          Methods

          The study population was comprised of 448 women with a previously abnormal Pap who were referred to the Barretos Cancer Hospital (Brazil) for diagnosis and treatment. Two cervical samples were collected immediately before colposcopy, one for the hr-HPV-DNA test and cytology and the other for the HPV16/18-E6 test using high-affinity monoclonal antibodies (mAb). Women with a histologic diagnosis of cervical intraepithelial neoplasia grade 2 or 3 were considered to be positive cases. Different strategies using a combination of screening methods (HPV-DNA) and triage tests (cytology and HPV16/18-E6) were also examined and compared.

          Results

          The HPV16/18-E6 test exhibited a lower positivity rate compared with the HPV-DNA test (19.0% vs. 29.3%, p<0.001) and a moderate/high agreement (kappa = 0.68, 95%CI: 0.60–0.75). It also exhibited a significantly lower sensitivity for CIN2+ and CIN3+ detection compared to the HPV-DNA test and a significantly higher specificity. The HPV16/18-E6 test was no different from cytology in terms of sensitivity, but it exhibited a significantly higher specificity in comparison to ASCH+. A triage test after HPV-DNA detection using the HPV16/18-E6 test exhibited a significantly higher specificity compared with a triage test of ASCH+ to CIN2+ (91.8% vs. 87.4%, p = 0.04) and CIN3+ (88.6% vs. 84.0%, p = 0.05).

          Conclusion

          The HPV16/18-E6 test exhibited moderate/high agreement with the HPV-DNA test but lower sensitivity and higher specificity for the detection of CIN2+ and CIN3+. In addition, its performance was quite similar to cytology, but because of the structural design addressed for the detection of HPV16/18-E6 protein, the test can miss some CIN2/3+ lesions caused by other high-risk HPV types.

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          Most cited references33

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          Natural history of cervical neoplasia and risk of invasive cancer in women with cervical intraepithelial neoplasia 3: a retrospective cohort study.

          The invasive potential of cervical intraepithelial neoplasia 3 (CIN3; also termed stage 0 carcinoma) has been poorly defined. At the National Women's Hospital, Auckland, New Zealand, treatment of CIN3 was withheld from a substantial number of women between 1965 and 1974 as part of an unethical clinical study. The resulting variation in management allows comparison of the long-term risk of invasive cancer of the cervix in women whose lesion was minimally disturbed with those who had adequate initial treatment followed by conventional management. We aimed to estimate the long-term risk of invasive cancer in these two groups of women. A judicial inquiry referred for independent clinical review in 1988 all women for whom there remained doubt about the adequacy of their management. Between February, 2001, and December, 2004, medical records, cytology, and histopathology were reviewed for all women with CIN3 diagnosed between 1955 and 1976, whose treatment was reviewed by judicial inquiry and whose medical records could be located, and linkages were done with cancer and death registers and electoral rolls. To take into account the probability that the CIN3 lesion had been completely removed, we classified adequacy of treatment by type of procedure, presence of CIN3 at the excision margin, and subsequent cytology. The primary outcome was cumulative incidence of invasive cancer of the cervix or vaginal vault. Follow-up continued until death or Dec 31, 2000, whichever came first. Analyses accounted for procedures during follow-up. 1229 women whose treatment was reviewed by the judicial inquiry in 1987-88 were included. Of these, 48 records (4%) could not be located and 47 women (4%) did not meet the inclusion criteria. At histopathological review, a further 71 (6% of 1134) women were excluded because the review diagnosis was not CIN3. We identified outcomes in the remaining 1063 (86% of 1229) women diagnosed with CIN3 at the hospital in 1955-76. In 143 women managed only by punch or wedge biopsy, cumulative incidence of invasive cancer of the cervix or vaginal vault was 31.3% (95% CI 22.7-42.3) at 30 years, and 50.3% (37.3-64.9) in the subset of 92 such women who had persistent disease within 24 months. However, cancer risk at 30 years was only 0.7% (0.3-1.9) in 593 women whose initial treatment was deemed adequate or probably adequate, and whose treatment for recurrent disease was conventional. This study provides the most valid direct estimates yet available of the rate of progression from CIN3 to invasive cancer. Women with untreated CIN3 are at high risk of cervical cancer, whereas the risk is very low in women treated conventionally throughout.
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            Mechanisms of human papillomavirus-induced oncogenesis.

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              Human papillomavirus testing in the prevention of cervical cancer.

              Strong evidence now supports the adoption of cervical cancer prevention strategies that explicitly focus on persistent infection with the causal agent, human papillomavirus (HPV). To inform an evidence-based transition to a new public health approach for cervical cancer screening, we summarize the natural history and cervical carcinogenicity of HPV and discuss the promise and uncertainties of currently available screening methods. New HPV infections acquired at any age are virtually always benign, but persistent infections with one of approximately 12 carcinogenic HPV types explain virtually all cases of cervical cancer. In the absence of an overtly persistent HPV infection, the risk of cervical cancer is extremely low. Thus, HPV test results predict the risk of cervical cancer and its precursors (cervical intraepithelial neoplasia grade 3) better and longer than cytological or colposcopic abnormalities, which are signs of HPV infection. The logical and inevitable move to HPV-based cervical cancer prevention strategies will require longer screening intervals that will disrupt current gynecologic and cytology laboratory practices built on frequent screening. A major challenge will be implementing programs that do not overtreat HPV-positive women who do not have obvious long-term persistence of HPV or treatable lesions at the time of initial evaluation. The greatest potential for reduction in cervical cancer rates from HPV screening is in low-resource regions that can implement infrequent rounds of low-cost HPV testing and treatment.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                20 October 2016
                2016
                : 11
                : 10
                : e0164892
                Affiliations
                [1 ]Molecular Oncology Research Center, Barretos Cancer Hospital/Pio XII Foundation, Barretos, Sao Paulo, Brazil
                [2 ]Pathology Department, Barretos Cancer Hospital/Pio XII Foundation, Barretos, Sao Paulo, Brazil
                [3 ]Cancer Prevention Department, Barretos Cancer Hospital/Pio XII Foundation, Barretos, Sao Paulo, Brazil
                [4 ]Department of Radiology and Oncology, Faculty of Medicine, University of Sao Paulo, Sao Paulo, Brazil
                [5 ]Virology Laboratory, LIM 52.The Institute of Tropical Medicine of the University of Sao Paulo, Sao Paulo, Brazil
                [6 ]Laboratory of Medical Research Laboratory, LIM 14.Department of Pathology, Faculty of Medicine, University of Sao Paulo, Sao Paulo, Brazil
                [7 ]Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal
                [8 ]ICVS/3B’s–PT Government Associate Laboratory, Braga/Guimaraes, Portugal
                Bharathidasan University, INDIA
                Author notes

                Competing Interests: The senior author (JHF) performed all analyses and states that the company did not interfere in the analyses or interpretation of the data or manuscript writing. The authors have declared that no competing interests exist.

                • Conceptualization: ATL CSN MDS JCPR MA LLV JEL ALF JHTGF.

                • Data curation: VSM JHTGF.

                • Formal analysis: VSM JHTGF.

                • Investigation: VSM ATL MDS JCPR MA.

                • Methodology: VSM ATL CSN MDS JCPR MA LLV JEL ALF JHTGF.

                • Project administration: JHTGF.

                • Supervision: JHTGF.

                • Writing – original draft: VSM ATL LLV ALF JHTGF.

                • Writing – review & editing: VSM ATL CSN MDS JCPR MA LLV JEL ALF JHTGF.

                Article
                PONE-D-16-25908
                10.1371/journal.pone.0164892
                5072685
                27764154
                3a9b9585-3a3f-41de-8f14-2aef4ea3bae9
                © 2016 Mariano et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 28 June 2016
                : 3 October 2016
                Page count
                Figures: 1, Tables: 6, Pages: 12
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100001807, Fundação de Amparo à Pesquisa do Estado de São Paulo;
                Award ID: 2012/51221-4
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001807, Fundação de Amparo à Pesquisa do Estado de São Paulo;
                Award ID: 2008/57889-1
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100003593, Conselho Nacional de Desenvolvimento Científico e Tecnológico;
                Award ID: 573799/2008-3
                Award Recipient :
                FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo to JHGF 2012/51221-4 and to LLV - 2008/57889-1, for HPV-DNA tests—Cobas test) and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico to LLV - 573799/2008-3). Arbor Vita Corporation donated all OncoE6 tests and no role in decision to publish.
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