scholarly journals Evaluation of DNA extraction protocols from liquid-based cytology specimens for studying cervical microbiota

2020 ◽  
Author(s):  
Takeo Shibata ◽  
Mayumi Nakagawa ◽  
Hannah N. Coleman ◽  
Sarah M. Owens ◽  
William W. Greenfield ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Dna Extraction ◽  
Cervical Cancer Screening ◽  
Intraepithelial Neoplasia ◽  
Extraction Protocol ◽  
Hpv Genotyping ◽  
Liquid Based Cytology

AbstractBackgroundCervical microbiota (CM) are considered an important factor affecting the progression of cervical intraepithelial neoplasia (CIN) and are implicated in the persistence of human papillomavirus (HPV). Collection of liquid-based cytology (LBC) samples is routine for cervical cancer screening and HPV genotyping, and can be used for long-term cytological biobanking. Herein, we investigate the feasibility of leveraging LBC specimens for use in CM surveys by amplicon sequencing. As methodological differences in DNA extraction protocols can potentially bias the composition of microbiota, we set out to determine the performance of four commonly used DNA extraction kits (ZymoBIOMICS DNA Miniprep Kit; QIAamp PowerFecal Pro DNA Kit; QIAamp DNA Mini Kit; and IndiSpin Pathogen Kit) and their ability to capture the diversity of CM from LBC specimens.ResultsLBC specimens from 20 patients (stored for 716 ± 105 days) with cervical intraepithelial neoplasia (CIN) 2/3 or suspected CIN2/3 were each aliquoted for the four kits. We observed that, regardless of the extraction protocol used, all kits provided equivalent accessibility to the cervical microbiome, with some minor differences. For example, the ZymoBIOMICS kit appeared to differentially increase access of several microbiota compared to the other kits. Potential kit contaminants were observed as well. Approximately 80% microbial genera were shared among all DNA extraction protocols. The variance of microbial composition per individual was larger than that of the DNA extraction protocol used. We also observed that HPV16 infection was significantly associated with community types that were not dominated by Lactobacillus iners.ConclusionsCollection of LBC specimens is routine for cervical cancer screening and HPV genotyping, and can be used for long-term cytological biobanking. We demonstrated that LBC samples, which had been under prolonged storage prior to DNA extraction, were able to provide a robust assessment of the CM and its relationship to HPV status, regardless of the extraction kit used. Being able to retroactively access the CM from biobanked LBC samples, will allow researchers to better interrogate historical interactions between the CM and its relationship to CIN and HPV. This alone has the potential to bring CM research one-step closer to the clinical practice.

scholarly journals Evaluation of DNA extraction protocols from liquid-based cytology specimens for studying cervical microbiota

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0237556
Author(s):  
Takeo Shibata ◽  
Mayumi Nakagawa ◽  
Hannah N. Coleman ◽  
Sarah M. Owens ◽  
William W. Greenfield ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Cancer Screening ◽  
Dna Extraction ◽  
Intraepithelial Neoplasia ◽  
Microbial Composition ◽  
Community Types ◽  
Liquid Based Cytology ◽  
Microbial Dna

Cervical microbiota (CM) are considered an important factor affecting the progression of cervical intraepithelial neoplasia (CIN) and are implicated in the persistence of human papillomavirus (HPV). Collection of liquid-based cytology (LBC) samples is routine for cervical cancer screening and HPV genotyping and can be used for long-term cytological biobanking. We sought to determine whether it is possible to access microbial DNA from LBC specimens, and compared the performance of four different extraction protocols: (ZymoBIOMICS DNA Miniprep Kit; QIAamp PowerFecal Pro DNA Kit; QIAamp DNA Mini Kit; and IndiSpin Pathogen Kit) and their ability to capture the diversity of CM from LBC specimens. LBC specimens from 20 patients (stored for 716 ± 105 days) with CIN values of 2 or 3 were each aliquoted for each of the four kits. Loss of microbial diversity due to long-term LBC storage could not be assessed due to lack of fresh LBC samples. Comparisons with other types of cervical sampling were not performed. We observed that all DNA extraction kits provided equivalent accessibility to the cervical microbial DNA within stored LBC samples. Approximately 80% microbial genera were shared among all DNA extraction protocols. Potential kit contaminants were observed as well. Variation between individuals was a significantly greater influence on the observed microbial composition than was the method of DNA extraction. We also observed that HPV16 was significantly associated with community types that were not dominated by Lactobacillus iners.

scholarly journals Effect of introducing human papillomavirus genotyping into real-world screening on cervical cancer screening in China: a retrospective population-based cohort study

2021 ◽  
Vol 13 ◽  
pp. 175883592110109
Author(s):  
Binhua Dong ◽  
Huachun Zou ◽  
Xiaodan Mao ◽  
Yingying Su ◽  
Hangjing Gao ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cohort Study ◽  
Human Papillomavirus ◽  
Cancer Screening ◽  
Cervical Cancer Screening ◽  
Cox Regression ◽  
Intraepithelial Neoplasia ◽  
Population Based ◽  
Hpv Genotyping ◽  
Cytology Screening

Background: China’s Fujian Cervical Pilot Project (FCPP) transitioned cervical cancer screening from high-risk human papillomavirus (HR-HPV) nongenotyping to genotyping. We investigated the clinical impact of this introduction, comparing performance indicators between HR-HPV genotyping combined with cytology screening (HR-HPV genotyping period) and the previous HR-HPV nongenotyping combined with cytology screening (HR-HPV nongenotyping period). Methods: A retrospective population-based cohort study was performed using data from the FCPP for China. We obtained data for the HR-HPV nongenotyping period from 1 January 2012 to 31 December 2013, and for the HR-HPV genotyping period from 1 January 2014 to 31 December 2016. Propensity score matching was used to match women from the two periods. Multivariable Cox regression was used to assess factors associated with cervical intraepithelial neoplasia of grade 2 or worse (CIN2+). The primary outcome was the incidence of CIN2+ in women aged ⩾25 years. Performance was assessed and included consistency, reach, effectiveness, adoption, implementation and cost. Results: Compared with HR-HPV nongenotyping period, in the HR-HPV genotyping period, more CIN2+ cases were identified at the initial screening (3.06% versus 2.32%; p < 0.001); the rate of colposcopy referral was higher (10.87% versus 6.64%; p < 0.001); and the hazard ratio of CIN2+ diagnosis was 1.64 (95% confidence interval, 1.43–1.88; p < 0.001) after controlling for health insurance status and age. The total costs of the first round of screening (US$66,609 versus US$65,226; p = 0.293) were similar during the two periods. Higher screening coverage (25.95% versus 25.19%; p = 0.007), higher compliance with age recommendations (92.70% versus 91.69%; p = 0.001), lower over-screening (4.92% versus 10.15%; p < 0.001), and reduced unqualified samples (cytology: 1.48% versus 1.73%, p = 0.099; HR-HPV: 0.57% versus 1.34%, p < 0.001) were observed in the HR-HPV genotyping period. Conclusions: Introduction of an HR-HPV genotyping assay in China could detect more CIN2+ lesions at earlier stages and improve programmatic indicators. Evidence suggests that the introduction of HR-HPV genotyping is likely to accelerate the elimination of cervical cancer in China.

Real time quantitative methylation detection of PAX1 gene in cervical cancer screening

2020 ◽  
Vol 30 (10) ◽  
pp. 1488-1492
Author(s):  
Haifeng Liu ◽  
Xia Meng ◽  
Jingyi Wang
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Real Time ◽  
Cervical Cancer Screening ◽  
Invasive Cervical Cancer ◽  
Intraepithelial Neoplasia ◽  
Cancer Group ◽  
Polymerase Chain ◽  
Folic Acid Receptor

IntroductionDNA methylation is currently found to be associated with the progression of cervical intraepithelial neoplasia and the development of cervical cancer. The aim of this study was to analyze the role of real time quantitative methylation detection of the PAX1 gene in cervical cancer screening.MethodsAll eligible patients who underwent multiple detections for cervical cancer were assigned to the normal cervical group (n=21), cervical intraepithelial neoplasia I group (n=7), cervical intraepithelial neoplasia II+III group (n=12), or invasive cervical cancer group (n=14) based on pathological gradings. The methylation level of the PAX1 gene was detected using the real time quantitative methylation specific polymerase chain reaction assay and assessed by △Cp value. The diagnostic performance of PAX1 methylation detection was compared with folic acid receptor mediated diagnosis, the Thinprep cytology test, and human papilloma virus (HPV) testing.ResultsThe △Cp value in the invasive cervical cancer group was (6.15±4.07), significantly lower than that in the other groups (F=26.45, p<0.001). The area under the curve (AUC) of PAX1 methylation detection was 0.902 (95% confidence interval (CI) 0.817–0.986; p<0.001), and sensitivity and specificity were 92.30% and 78.60% when the cut-off value of △Cp was 13.28. The AUC of PAX1 methylation detection was notably larger compared with 0.709 for folic acid receptor mediated diagnosis (95% CI 0.568–0.849, p=0.009), 0.702 for the Thinprep cytology test (95% CI 0.559–0.844, p=0.015), and 0.655 for HPV testing (95% CI 0.508–0.802, p=0.014).ConclusionThrough quantitative methylation specific polymerase chain reaction assay characterized by rapid screening and simple operation, the methylation detection of the PAX1 gene exhibited a higher diagnostic performance and may be a promising method for cervical cancer screening.

scholarly journals Guidelines for HPV-DNA Testing for Cervical Cancer Screening in Brazil

2018 ◽  
Vol 40 (06) ◽  
pp. 360-368 ◽  
Author(s):  
Luiz Zeferino ◽  
Joana Bastos ◽  
Diama Vale ◽  
Rita Zanine ◽  
Yara Melo ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Cervical Cancer Screening ◽  
Residual Disease ◽  
Intraepithelial Neoplasia ◽  
Low Grade ◽  
Hpv Dna ◽  
Published Evidence ◽  
Hpv Dna Tests

AbstractEvidence-based clinical guidelines ensure best practice protocols are available in health care. There is a widespread use of human papillomavirus deoxyribonucleic acid (HPV-DNA) tests in Brazil, regardless of the lack of official guidelines. On behalf of the Brazilian Association for the Lower Genital Tract Pathology and Colposcopy (ABPTGIC, in the Portuguese acronym), a team of reviewers searched for published evidence and developed a set of recommendations for the use of HPV-DNA tests in cervical cancer screening in Brazil. The product of this process was debated and consensus was sought by the participants. One concern of the authors was the inclusion of these tests in the assessment of women with cytologic atypia and women treated for cervical intraepithelial neoplasia (CIN). Testing for HPV is recommended in an organized screening scenario to identify women with precursor lesions or asymptomatic cervical cancer older than 30 years of age, and it can be performed every 5 years. It also has value after the cytology showing atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesions (LSILs) as a triage test for colposcopy, in the investigation of other cytological alterations when no abnormal findings are observed at colposcopy, seeking to exclude disease, or, further, after treatment of high-grade cervical intraepithelial neoplasia, to rule out residual disease.

scholarly journals The Vaginal Microbiome and Cervical Cancer Screening in Low- and Middle-Income Countries

2019 ◽  
Vol 5 (Supplement_1) ◽  
pp. 6-6
Author(s):  
Supriya D. Mehta
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Cervical Cancer Screening ◽  
Intraepithelial Neoplasia ◽  
Vaginal Microbiome ◽  
Middle Income ◽  
Middle Income Countries ◽  
Cancer Pathogenesis ◽  
Low And Middle Income

PURPOSE Globally, cervical cancer affects more than a half-million women each year, with disproportionate incidence and mortality for women in low- and middle-income countries. Early detection through cervical cancer screening saves lives but is hampered by poor coverage, suboptimal detection accuracy, and lack of access to and delays in effective treatment. METHODS Emerging evidence that indicates how the vaginal microbiome can modify progression of human papillomavirus (HPV) infection and cervical cancer pathogenesis is surveyed. This presentation features a discussion of how the vaginal microbiome may affect cervical cancer screening and how cervical cancer screening may incorporate vaginal microbiome health in low- and middle-income countries. RESULTS Vaginal dysbiosis as a clinical syndrome may be called bacterial vaginosis (BV), a condition that represents a shift from a Lactobacillus-dominant vaginal microbiome to one that is polymicrobial and often associated with increased mucosal inflammation. Meta-analyses and prospective studies demonstrate an association between vaginal dysbiosis and increased risk of HPV incidence and persistence and high-grade lesions and cancer. Increasing vaginal microbiome diversity is associated with progression of cervical intraepithelial neoplasia. Vaginal microbiota that are associated with greater likelihood of HPV detection in molecular studies are also commonly associated with BV. There are numerous challenges to incorporating microbiome measurement in population-level cervical cancer screening and unanswered research questions on its immediate utility. BV may serve as a measure of vaginal microbiome health, although there are no guidelines or recommendations for regular BV screening and treatment. CONCLUSION Ongoing and planned longitudinal studies should evaluate BV screening in association with high-risk HPV, results of cervical cancer screening, and progression of cervical intraepithelial neoplasia to assess the utility of BV screening and treatment as an adjunct to cervical cancer screening and potential intervention.

scholarly journals Expanding the upper age limit for cervical cancer screening: a protocol for a nationwide non-randomised intervention study

BMJ Open ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. e039636
Author(s):  
Mette Tranberg ◽  
Lone Kjeld Petersen ◽  
Klara Miriam Elfström ◽  
Anne Hammer ◽  
Jan Blaakær ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Cancer Screening ◽  
Intervention Study ◽  
Reference Group ◽  
Intervention Group ◽  
Intraepithelial Neoplasia ◽  
Cervical Cytology ◽  
Cervical Intraepithelial Neoplasia Grade

IntroductionCervical cancer screening ceases between the ages of 60 and 65 in most countries. Yet, a relatively high proportion of cervical cancers are diagnosed in women above the screening age. This study will evaluate if screening of women aged 65–69 years may result in increased detection of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) compared with women not invited to screening. Invited women may choose between general practitioner (GP)-based screening or cervico-vaginal self-sampling. Furthermore, the study will assess if self-sampling is superior to GP-based screening in reaching long-term unscreened women.Methods and analysisThis population-based non-randomised intervention study will include 10 000 women aged 65–69 years, with no record of a cervical cytology sample or screening invitation in the 5 years before inclusion. Women who have opted-out of the screening programme or have a record of hysterectomy or cervical amputation are excluded. Women residing in the Central Denmark Region (CDR) are allocated to the intervention group, while women residing in the remaining four Danish regions are allocated to the reference group. The intervention group is invited for human papillomavirus-based screening by attending routine screening at the GP or by requesting a self-sampling kit. The reference group receives standard care which is the opportunity to have a cervical cytology sample obtained at the GP or by a gynaecologist. The study started in April 2019 and will run over the next 4.5 years. The primary outcome will be the proportion of CIN2+ detected in the intervention and reference groups. In the intervention group, the proportion of long-term unscreened women attending GP-based screening or self-sampling will be compared.Ethics and disseminationThe study has been submitted to the Ethical Committee in the CDR which deemed that the study was not notifiable to the Committee and informed consent is therefore not required. The study is approved by the Danish Data Protection Regulation and the Danish Patient Safety Authority. Results will be disseminated in peer-reviewed journals.Trial registration numberNCT04114968.

scholarly journals Co-infection with trichomonas vaginalis increases the risk of cervical intraepithelial neoplasia grade 2-3 among HPV16 positive female: a large population-based study.

2020 ◽  
Author(s):  
Mei Yang ◽  
Lin Li ◽  
Chunfan Jiang ◽  
Xiaomin Qin ◽  
Min Zhou ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Cervical Cancer Screening ◽  
Large Population ◽  
Intraepithelial Neoplasia ◽  
Central China ◽  
Cervical Lesions ◽  
Hpv 16 ◽  
Hpv 18

Abstract Background: Evidence suggested that vaginal microbiome played a functional role in the progression of cervical lesions in female infected by HPV. This study aimed at evaluating the influence of common vaginal infection on the carcinogenicity of hr-HPV. Methods: From January 15, 2017 to December 31, 2017, 310,545 female aged at least 30 years old had been recruited for cervical cancer screening from 9 clinical research centers in Central China. All the recruited participants received hr-HPV genotyping for cervical cancer screening and vaginal microenvironment test by a high vaginal swab. Colposcopy-directed biopsy was recommended for female who were infected with HPV 16 and HPV 18, and other positive hr-HPV types through test had undertaken triage using liquid-based cytology, cases with the results ≥ ASCUS among them were referred to colposcopy directly, and cervical tissues were taken for pathology examination to make clear the presence or absence of other cervical lesions.Results: Among 310,545 female, 6,067 (1.95%) were tested with positive HPV 16 and HPV 18, 18,297 (5.89%) were tested with other positive hr-HPV genotypes, cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3 and invasive cervical cancer (ICC) were detected in 861 cases, 377 cases, 423 cases, and 77 cases, respectively. Candida albicans and Gardnerella were not associated with the detection of cervical lesions. Positive trichomonas vaginitis (TV) was correlated with hr-HPV infection (p<0.0001). Co-infection with TV increased the risk of CIN 1 among female infected with hr-HPV (OR 1.18, 95% CI: 1.42-2.31). Co-infection with TV increased the risk of CIN 2-3 among female infected with HPV 16 (OR 1.71, 95% CI: 1.16-2.53).Conclusions: Co-infection of TV and HPV 16 is a significant factor for the detection of cervical lesions.

scholarly journals Co-infection with trichomonas vaginalis increases the risk of cervical intraepithelial neoplasia grade 2-3 among HPV16 positive female: a large population-based study.

2020 ◽  
Author(s):  
Mei Yang ◽  
Lin Li ◽  
Chunfan Jiang ◽  
Xiaomin Qin ◽  
Min Zhou ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Cervical Cancer Screening ◽  
Large Population ◽  
Intraepithelial Neoplasia ◽  
Central China ◽  
Cervical Lesions ◽  
Hpv 16 ◽  
Hpv 18

Abstract Background: Evidence suggested that vaginal microbiome played a functional role in the progression of cervical lesions in female infected by HPV. This study aimed at evaluating the influence of common vaginal infection on the carcinogenicity of high risk HPV (hr-HPV). Methods: From January 15, 2017 to December 31, 2017, 310,545 female aged at least 30 years old had been recruited for cervical cancer screening from 9 clinical research centers in Central China. All the recruited participants received hr-HPV genotyping for cervical cancer screening and vaginal microenvironment test by a high vaginal swab. Colposcopy-directed biopsy was recommended for female who were infected with HPV 16 and HPV 18, and other positive hr-HPV types through test had undertaken triage using liquid-based cytology, cases with the results ≥ ASCUS among them were referred to colposcopy directly, and cervical tissues were taken for pathology examination to make clear the presence or absence of other cervical lesions.Results: Among 310,545 female, 6,067 (1.95%) were tested with positive HPV 16 and HPV 18, 18,297 (5.89%) were tested with other positive hr-HPV genotypes, cervical intraepithelial neoplasia (CIN) 1, CIN 2, CIN 3 and invasive cervical cancer (ICC) were detected in 861 cases, 377 cases, 423 cases, and 77 cases, respectively. Candida albicans and Gardnerella were not associated with the detection of cervical lesions. Positive trichomonas vaginitis (TV) was correlated with hr-HPV infection (p<0.0001). Co-infection with TV increased the risk of CIN 1 among female infected with hr-HPV (OR 1.18, 95% CI: 1.42-2.31). Co-infection with TV increased the risk of CIN 2-3 among female infected with HPV 16 (OR 1.71, 95% CI: 1.16-2.53).Conclusions: Co-infection of TV and HPV 16 is a significant factor for the detection of cervical lesions.

Sign in / Sign up

Export Citation Format

Share Document