Combining HPV DNA load with p16/Ki-67 staining to detect cervical precancerous lesions and predict the progression of CIN1–2 lesions

Abstract Background Human Papilloma Virus (HPV) DNA tests are highly sensitive and can triage women with mild lesions, improving the prognosis and diagnosis of cervical lesions. However, additional efficient strategies should be developed to improve the specificity of these tests. Methods This study aimed to evaluate the clinical value of HPV DNA load in improving the diagnosis and prognosis of cervical lesions by p16/Ki-67 testing. Histological samples were collected from 350 women with HR-HPV genotyping and analyzed by qRT-PCR. Immunohistochemical staining was used to assess p16 and Ki-67 expression and clinical performance characteristics were calculated. Results Of the cases, 271 had detectable HR-HPV infection, in which HPV-16 was most prevalent (52.0%), followed by HPV-58 (22.5%). P16/Ki-67-positivity increased with histological severity but not for HR-HPV infection. Amongst the 13 HR-HPV genotypes, only HPV-16 (P = 0.016) and HPV-58 (P = 0.004) viral loads significantly correlated with lesion severity. The P16/Ki-67/HPV DNA load co-test indicated an increased sensitivity for the detection of cervical intraepithelial neoplasia (CIN) lesions compared to p16/Ki-67 staining in HPV-16 and/or 58 positive cases. Viral load did not improve the sensitivity of p16/Ki-67 co-test in non-HPV-16 or 58 positive cases. The clinical performance of the p16/Ki-67/HPV DNA load co-test was limited for the prediction of the outcome of CIN1 lesions. However, amongst the 12 HPV-16 and/or 58 positive CIN2 cases in which return visit results were obtained, the behavior of the lesions could be predicted, with a sensitivity, specificity, positive prediction rate (PPV), and negative prediction rate (NPV) of 0.667, 1, 1 and 0.5, respectively. Conclusion Combination of the assessment of HPV DNA load with the intensity of p16 and Ki-67 staining could increase the sensitivity of CIN lesion diagnosis and predict the outcome of CIN2 in patients with a HPV-16 and/or 58 infection.

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State of HPV16 integration in Lithuanian women with cervical neoplasia

Open Medicine ◽

10.2478/s11536-011-0004-6 ◽

2011 ◽

Vol 6 (2) ◽

pp. 205-212

Author(s):

Agne Sepetiene ◽

Zivile Gudlevicienė ◽

Zana Bumbuliene ◽

Grazina Drasutiene ◽

Janina Didziapetriene

Keyword(s):

Cervical Cancer ◽

Hpv Infection ◽

Rapid Progression ◽

The European Union ◽

Cervical Lesions ◽

Hpv 16 ◽

Hpv Dna ◽

Cin Iii ◽

Cell Genome ◽

Virus Integration

AbstractCervical cancer morbidity and mortality in Lithuania is one of the biggest in the European Union. The main risk factor of cervical cancer is human papillomavirus (HPV). The deletion of the HPV E2 gene influences HPV DNA integration into the cell genome, as well as a rapid progression of cervical lesions. The purpose of this study is to determine HPV, its types, and HPV 16 integration in different grades of cervical intraepithelial neoplasias (CIN). 253 women with cytological lesions were involved in the study. After a histology, 31 women were diagnosed with CIN I, 35 with CIN II, and 51 with CIN III. The biggest prevalence of HPV infection was detected in women younger than 25 years old (69.7%) and in women with CIN II (90.9%). HPV 16 was detected in 67.8% of all cases, with the highest prevalence in CIN III (84.4%). A partial integration form was detected in 65.0% of HPV 16 infected women, a complete virus integration in 26.5%, and an episomal form in 8.4% of cases. Our study concludes that in all the cases confirmed using a histology, the partial virus integration form of CIN was identified the most. It was less frequently detected in CIN I cases (60.0%), but more frequently in CIN II and CIN III cases (72.8 and 69.3%, respectively).

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Human Papilloma Virus L1 Gene Methylation as a Potential Biomarker for Precancerous Cervical Lesion: a Preliminary Report

Indonesian Journal of Obstetrics and Gynecology ◽

10.32771/inajog.v5i2.532 ◽

2017 ◽

pp. 120

Author(s):

Suzanna P Mongan ◽

Andrijono Andrijono ◽

Hartono Tjahadi

Keyword(s):

Cervical Cancer ◽

Precancerous Lesions ◽

Bisulfite Conversion ◽

Cervical Lesions ◽

Gene Methylation ◽

Hpv 16 ◽

Hpv Dna ◽

Precancerous Cervical Lesions ◽

Hpv 18 ◽

Thin Band

Objective: To determine whether HPV L1 gene methylation can be used in triage of precancerous cervical lesions. The main objective is to determine the genotype of HPV in cervical precancerous lesions and to determine the percentage, the sensitivity, specificity, positive predictive value, negative predictive value, and likelihood ratio of DNA HPV L1 methylation in precancerous cervical lesions. Methods: A number of 57 samples of paraffin blocks (FFPE) from precancerous lesions and cervical cancer biopsies in the Department of Pathology Faculty of Medicine-Cipto Mangunkusumo General Hospital that had been re-evaluated by the pathologist, underwent extraction of HPV DNA. The genotypes of HPV DNA were examined using primers GP5 / 6 and specific HPV 16, HPV 18 and HPV 52 probes and analyzed by real time PCR. Sequencing was performed on samples with unknown HPV DNA type that were detected using the specific probes to determine the type of HPV. Bisulfite conversion procedure was then performed for the samples that met the inclusion criteria. Results: There were 30 samples (52.6%) with CIN 1, 12 samples (21.1%) CIN 2, 9 samples (15.8%) CIN 3 and 6 samples (10.5%) of cervical cancer. Most of the samples were 36-45 years (35.1%). Of the total 57 samples, 55 samples were successfully extracted and determined the DNA genotyping of HPV (96.5%). HPV 16 infections both in the form of single or multiple was found to be 76.36%. The samples were mostly dominated by co-infection of HPV16 and 18 (49.1%) followed by HPV 16 (24.6%) and HPV 18 (14.0%). Based on the sequencing results there were other types of high risk HPV infection found: HPV 33, HPV 35, HPV 58 and also undeterminate risk HPV 53 and low risk HPV 54. After several procedures of optimization for methylation examination of HPV DNA L1 there was thin band found in electrophoresis procedure in 8 of 42 samples (19%) of HPV 16 after bisulfite conversion but once it was purified there weren’t any band found so we can not proceed to the stage sequencing. Until now we are still in the stage of optimizing the methylation procedure. Conclusion: HPV 16 infection were most commonly found in the form of single or multiple. Co-infection of HPV 16 and 18 were found in the majority of the samples. There were no significant correlation between HPV type and the severity of cervical lesions. Until now, the examination of DNA methylation HPV L1 already obtained eight samples of HPV 16 with a thin band on electrophoresis but the result could not be concluded because it is still in the process of optimization. [Indones J Obstet Gynecol 2017; 5-2: 120-126] Keywords: HPV DNA genotype, L1 gene methylation, precancerous cervical lesions

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Prevalence of human papillomavirus genotype among Moroccan women during a local screening program

The Journal of Infection in Developing Countries ◽

10.3855/jidc.781 ◽

2010 ◽

Vol 4 (11) ◽

pp. 732-739 ◽

Cited By ~ 14

Author(s):

Zaitouna Alhamany ◽

Mohammed El Mzibri ◽

Aicha Kharbach ◽

Abderrahman Malihy ◽

Redouane Abouqal ◽

...

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Hpv Infection ◽

Dna Testing ◽

Cervical Lesions ◽

Abnormal Cytology ◽

Hpv 16 ◽

Hpv Dna ◽

Hpv Dna Testing ◽

Moroccan Women

Introduction: Many studies have indicated a causal relationship between genital human papillomavirus (HPV) infections and cervical cancer. This study aimed to determine the prevalence and genotypes of six high-risk oncogenic human papillomaviruses in cervical lesions from Moroccan women with normal and abnormal cytology. Methodology: The study included 938 women from the Children's and Mothers' Pathology Department of Ibn Sina Hospital, Rabat. Cytopathology examination was done by routine PAP smear testing. HPV DNA testing was conducted using DNA amplification by Polymerase Chain Reaction with subsequent typing by hybridization with specific probes for HPV types 16, 18, 31, 33, 35 and 45. Results: Cytopathology testing showed that only 16.3 % had an abnormal cytology, with a predominance of atypical squamous cell of undetermined significance (ASCUS) cases. The overall HPV prevalence was 15.7%. According to the cytology results, HPV infection was detected in 15.8% of normal and 14.38% of abnormal cases. Specific HPV genotyping showed a predominance of HPV 16 and 18. Double infection (HPV 16 + 18) was found in two cases whereas multiple infections (HPV 16+18+31) were detected in only one case. Evaluation of the relationship between HPV status and some environmental risk factors, including individual, socio-economic, and hygiene status, showed a significant association between HPV infection and oral contraceptive use. Conclusion: Based on these data, a combination of cytology and HPV DNA testing allows for identification of patients with a high risk of developing high-grade cervical lesions and improves cervical cancer prevention.

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Evaluation of Dynamic Thiol-Disulfide Homeostasis on Hpv Positive-Women in Progression to Cervical Intraepithelial Lesion

10.21203/rs.3.rs-559471/v1 ◽

2021 ◽

Author(s):

Recep Erin ◽

Yeşim Bayoğlu Tekin ◽

Hatice Küçük ◽

Özcan Erel

Keyword(s):

Precancerous Lesions ◽

Study Groups ◽

Systemic Circulation ◽

Hpv Infection ◽

Cross Sectional Study ◽

Cervical Lesion ◽

Cervical Lesions ◽

Cross Sectional ◽

Hpv Dna ◽

Total Thiol

Abstract PURPOSE: Dynamic thiol disulfide homeostasis (TDH) is critical in cervical carcinogenesis at HPV infection as a sign of antioxidant consumption native and total thiol levels decrease in progress to cervical intraepithelial lesions. TDH is the main actor in signaling pathways, apoptosis, antioxidant and detoxification reactions. In this study, we aimed to evaluate the effect of TDH intraepithelial progression of cervical precancerous lesions on HPV positive women.METHODS: This was a prospective cross-sectional study. Subjects were selected from newly diagnosed high risk HPV DNA-positive patients. TDH results were calculated as the levels of disulfide, native and total thiol, the ratios of disulfide/total thiol (SS/SH+SS), disulfide/native thiol (SS/SH) and native thiol/total thiol (SH/SH+SS).RESULTS: A total of 146 women were included in the study. Study groups were as group one; control included 66 participants, group two; HPV DNA-positive women without preinvasive cervical lesion included 30 participants and group three; HPV DNA-positive women with preinvasive cervical lesion included 50 participants. Native and total thiol levels were elevated on HPV-positive women without preinvasive cervical lesions. There were no significant differences between groups related to the ratios of SS/SH, SS/ Total SH, SH/ Total SH levels. CONCLUSIONS: HPV infection related to oxidative stress has effects on oxidant/antioxidant balance and could be demonstrated in systemic circulation by TDH parameters. Consumption of thiol substances play role in the cervical neoplastic process, replacement with antioxidants would be a treatment option for HPV infections.

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HPV 16 Is Related to the Progression of Cervical Intraepithelial Neoplasia Grade 2: A Case Series

Obstetrics and Gynecology International ◽

10.1155/2013/328909 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5 ◽

Cited By ~ 3

Author(s):

Maria Gabriela Loffredo D’Ottaviano ◽

Michelle Garcia Discacciati ◽

Maria Antonieta Andreoli ◽

Maria Cecília Costa ◽

Lara Termini ◽

...

Keyword(s):

Blot Hybridization ◽

Case Series ◽

Intraepithelial Neoplasia ◽

Hpv Infection ◽

Low Grade ◽

Cervical Smear ◽

Cervical Intraepithelial Neoplasia Grade ◽

Hpv 16 ◽

Hpv Dna ◽

Hpv Test

Purpose. To describe the acquisition, persistence, and clearance of HPV infection in women with CIN 2 followed up for 12 months.Methods. Thirty-seven women with CIN 2 biopsy, who have proven referral to cervical smear showing low-grade squamous intraepithelial lesions or atypical squamous cells of undetermined significance and tested for HPV, were followed up for one year with cervical smear, colposcopy, and HPV test every three months. HPV DNA was detected by the polymerase chain reaction and genotyping by reverse line blot hybridization assay.Results. CIN 2 regression rate was 49% (18/37), persistence as CIN 1 or CIN 2 was 22% (8/37), and progression to CIN 3 was 29% (11/37). Multiple HPV types were observed at admission in 41% (15/37) of cases. HPV 16 was detected at admission in 58% (11/19) of the cases that persisted/progressed and in 39% (7/18) of the cases that regressed. HPV 16 was considered possibly causal in 67% (10/15) of the cases that persisted or progressed and in 10% (1/10) of the cases that regressed (P=0.01).Conclusion. Multiple HPV infections were frequently detected among women with CIN 2 at admission and during the followup. The CIN 2 associated with HPV 16 was more likely to persist or to progress to CIN 3.

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Epidemiologic correlates of antibody response to human papillomavirus among women at low risk of cervical cancer

International Journal of STD & AIDS ◽

10.1258/095646203321264872 ◽

2003 ◽

Vol 14 (4) ◽

pp. 258-265 ◽

Cited By ~ 21

Author(s):

B Nonnenmacher ◽

J Pintos ◽

M C Bozzetti ◽

I Mielzinska-Lohnas ◽

A T Lorincz ◽

...

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Humoral Response ◽

Hpv Infection ◽

Low Risk ◽

Enzyme Linked Immunosorbent Assay ◽

Serological Response ◽

Cervical Lesions ◽

Hpv 16 ◽

Hpv Dna

A population at low risk for developing cervical cancer in Southern Brazil was studied to identify the main determinants of serological response to human papillomavirus (HPV). Enzyme-linked immunosorbent assay tests were performed in 976 women to detect serum IgG antibodies against HPV 16 L1 virus-like particles (VLPs) and HPVs 16, 18, 6 and 11 L1 VLPs as a mixture of antigens. Women with four or more sexual partners were more likely to be seropositive than women with one partner (HPV 16 serology odds ratio [OR]=3.06, 95% confidence interval [CI]: 2.0-4.8; HPV 6/11/16/18 serology OR=4.64, 95% CI: 3.0-7.2). HPV DNA and both serological responses were associated. Those positives to HPV 16 serology were twice as likely to have a cytological diagnosis of squamous intraepithelial lesions (SILs) than seronegatives (OR=2.07; 95% CI: 1.0-4.5, and OR=1.73; 95% CI: 0.8-3.8). Seropositivity to HPV 16 and HPV 6/11/16/18 antigens seem to be better markers of past sexual activity than current HPV infection, and humoral response to HPV 16 or HPV 6/11/16/18 may not be a strong indicator of cervical lesions in populations at low risk for cervical lesions.

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Preliminary Study on the Expression of Testin, p16 and Ki-67 in the Cervical Intraepithelial Neoplasia

Biomedicines ◽

10.3390/biomedicines9081010 ◽

2021 ◽

Vol 9 (8) ◽

pp. 1010

Author(s):

Aneta Popiel ◽

Aleksandra Piotrowska ◽

Patrycja Sputa-Grzegrzolka ◽

Beata Smolarz ◽

Hanna Romanowicz ◽

...

Keyword(s):

Cervical Cancer ◽

Precancerous Lesions ◽

Clinical Stage ◽

Intraepithelial Neoplasia ◽

Hpv Infection ◽

Control Group ◽

Ki 67 ◽

Cervical Precancerous Lesions ◽

P16 Expression

Cervical cancer is one of the most common malignant cancers in women worldwide. The 5-year survival rate is 65%; nevertheless, it depends on race, age, and clinical stage. In the oncogenesis of cervical cancer, persistent HPV infection plays a pivotal role. It disrupts the expression of key proteins as Ki-67, p16, involved in regulating the cell cycle. This study aimed to identify the potential role of testin in the diagnosis of cervical precancerous lesions (CIN). The study was performed on selected archival paraffin-embedded specimens of CIN1 (31), CIN2 (75), and CIN3 (123). Moderate positive correlation was observed between testin and Ki-67 as well as testin and p16 expression in all dysplastic lesions (r = 0.4209, r = 0.5681; p < 0.0001 for both). Statistical analysis showed stronger expression of the testin in dysplastic lesions vs. control group (p < 0.0001); moreover, expression was significantly higher in HSIL than LSIL group (p < 0.0024). In addition, a significantly stronger expression of testin was observed in CIN3 vs. CIN1 and CIN3 vs. CIN2. In our study, expression of Ki-67, p16, and testin increased gradually as the lesion progressed from LSIL to HSIL. The three markers complemented each other effectively, which may improve test sensitivity and specificity when used jointly.

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Determination of papilloma-virus infection reproductive phase in algorithm of monitoring patients with cervical displasia

Journal of obstetrics and women s diseases ◽

10.17816/jowd91125 ◽

2021 ◽

Vol 51 (4) ◽

pp. 61-63

Author(s):

S. A. Selkov ◽

G. N. Vedeneeva ◽

I. A. Baskakova ◽

S. R. Baur

Keyword(s):

Intraepithelial Neoplasia ◽

Hpv Infection ◽

Rt Pcr ◽

Chain Reaction ◽

Hpv 16 ◽

Papilloma Virus ◽

Hpv Dna ◽

Virus Activity ◽

Polymerase Chain

HPV 16 and 18 are known to be the main cause of cervical intraepithelial neoplasia (CIN) and cervical cancer. The terms of HPV persistence in the host and, coordinately, the risk of cervical neoplasia development and progression are determined in much extent by virus activity. The purpose of this investigation was the detection of HPV DNA presence in cervical epithelium as well as confirmation of its activity by means of immunocytochemistry and reverse transcriptase polymerase chain reaction. The level of HPV inf ection by oncogenic and nononcogenic types in 181 women with different cervical pathology was 55,8%. The active stage of HPV infection was confirmed in 27,5% of HPV-inf ected women mainly with low grades of CIN. The proof of reproductive general HPV infection was more informative with RT PCR just as for HPV 16 and 18 immunocytochemistry and RT PCR completed each anothe.

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Efficacy of HPV vaccination among seropositive, DNA negative cohorts.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.10552 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. 10552-10552

Author(s):

Colm Mac Eochagain ◽

Robert Power ◽

Imelda Parker

Keyword(s):

Persistent Infection ◽

Meta Analysis ◽

Hpv Vaccination ◽

Intraepithelial Neoplasia ◽

Hpv Infection ◽

Eligibility Criteria ◽

Hpv 16 ◽

Hpv Dna ◽

Dna Evidence ◽

Relative Risks

10552 Background: Prophylactic HPV vaccination of naïve cohorts is known to be highly effective in the prevention of incident HPV infection and HPV-associated cervical premalignancy. Conversely, vaccination of women with active (DNA+) HPV infections has been demonstrated to be ineffective. Vaccine efficacy among previously exposed, but currently uninfected women, i.e. those who have serological evidence of a prior HPV infection without corresponding detectable HPV DNA, remains incompletely defined. This meta-analysis assessed the serotype-specific efficacy of prophylactic HPV vaccination against HPV16/18 persistent infection (PI) and cervical intraepithelial neoplasia (CIN) among seropositive, DNA negative (SPDN) women enrolled to RCTs of HPV L1-based vaccines. Methods: The study protocol was prospectively registered on PROSPERO (CRD42020206888). Searches were conducted on 08/16/20 on MEDLINE, EMBASE, SCOPUS and CENTRAL. RCTs of L1-based prophylactic bivalent or quadrivalent HPV vaccines, reporting serotype-specific clinical efficacy endpoints in the HPV16/18 seropositive, DNA-negative populations were included. Two authors independently screened studies, extracted data and assessed for bias. Data for SPDN women were extracted from subgroup analyses within primary and secondary publications, publication supplements, and manufacturers' clinical study reports. Relative risks (RR) of 6-month persistent infection (6mPI), 12-month persistent infection (12mPI), CIN1+ and CIN2+ were pooled using a random-effects model. Results: A total of 1727 citations were reviewed. 8 studies, with a total of 9569 SPDN participants, met all eligibility criteria. The relative risk of 6mPI (RR: 0.22, 95% CI 0.08-0.61, p = 0.018), 12mPI (RR: 0.20, 95% CI 0.05-0.80, p = 0.035), CIN1+ (RR:0.13, 95% CI 0.05-0.30, p = 0.003) and CIN2+ (RR: 0.15, 95% CI 0.04-0.59, p = 0.022) was significantly reduced in the vaccinated compared to the unvaccinated group. The number needed to vaccinate (NNV) to prevent one case of CIN1+ and CIN2+ was 152 and 208 respectively. Conclusions: Our findings suggest high serotype-specific efficacy for HPV vaccination among cohorts of women with evidence of prior HPV 16/18 infections. Women without DNA evidence of active infection may be seronegative, implying naïve status and > 99% efficacy; or seropositive, implying SPDN status and 87% efficacy against CIN1+ for HPV 16/18. Women without DNA evidence of HPV 16/18 infection should be offered prophylactic HPV vaccination regardless of prior exposure history.

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Comparison of Onclarity Human Papillomavirus (HPV) Assay with Hybrid Capture II HPV DNA Assay for Detection of Cervical Intraepithelial Neoplasia Grade 2 and 3 Lesions

Journal of Clinical Microbiology ◽

10.1128/jcm.00246-15 ◽

2015 ◽

Vol 53 (7) ◽

pp. 2109-2114 ◽

Cited By ~ 11

Author(s):

F. Bottari ◽

M. Sideri ◽

C. Gulmini ◽

S. Igidbashian ◽

A. Tricca ◽

...

Keyword(s):

Human Papillomavirus ◽

Cervical Intraepithelial Neoplasia ◽

Clinical Performance ◽

Absolute Risk ◽

Intraepithelial Neoplasia ◽

Cervical Lesions ◽

Cervical Intraepithelial Neoplasia Grade ◽

Hybrid Capture ◽

Dna Test ◽

Hpv Dna

Analytical and clinical performance validation is essential before introduction of a new human papillomavirus (HPV) assay into clinical practice. This study compares the new BD Onclarity HPV assay, which detects E6/E7 DNA from 14 high-risk HPV types, to the Hybrid Capture II (HC2) HPV DNA test, to concurrent cytology and histology results, in order to evaluate its performance in detecting high-grade cervical lesions. A population of 567 women, including 325 with ≥ASCUS (where ASCUS stands for atypical cells of undetermined significance) and any HC2 result and 242 with both negative cytology and negative HC2 results, were prospectively enrolled for the study. The overall agreement between Onclarity and HC2 was 94.6% (95% confidence intervals [CI], 92.3% to 96.2%). In this population with a high prevalence of disease, the relative sensitivities (versus adjudicated cervical intraepithelial neoplasia grades 2 and 3 [CIN2+] histology endpoints) of the Onclarity and HC2 tests were 95.2% (95% CI, 90.7% to 97.5%) and 96.9% (95% CI, 92.9% to 98.7%), respectively, and the relative specificities were 50.3% (95% CI, 43.2% to 57.4%) for BD and 40.8% (95% CI, 33.9%, 48.1%) for HC2. These results indicate that the BD Onclarity HPV assay has sensitivity comparable to that of the HC2 assay, with a trend to an increased specificity. Moreover, as Onclarity gives the chance to discriminate between the different genotypes, we calculated the genotype prevalence and the absolute risk of CIN2+: HPV 16 was the most prevalent genotype (19.8%) with an absolute risk of CIN2+ of 77.1%.

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