Positive hrHPV Test Research Articles

Population-based cervical cancer screening using high-risk HPV DNA test and liquid-based cytology in northern Thailand.

Northern Thailand is a region with a high cervical cancer incidence. Combined high-risk HPV (hrHPV) DNA testing and cytology (co-testing) has increasingly gained acceptance for cervical cancer screening. However, to our knowledge, data from a population-based screening using co-testing have not been available in this region. This study therefore aimed to evaluate the performance of cytology and hrHPV test in women in northern Thailand. Cervical samples were collected for hybrid capture 2 (HC2) testing and liquid-based cytology from women aged 30 to 60 years who were residents in 3 prefectures of Chiang Mai in northern Thailand between May and September 2011. Women with positive cytology were referred to colposcopy, while women with positive for HC2 only were followed for 2 years. Of 2,752 women included in this study, 3.0% were positive in both tests, 4.1% for HC2 only, and 1.3% had positive cytology only. At baseline screening, positive HC2 was observed in 70.6% among cytology-positive women compared with 4.3% among cytology-negative women. The prevalence of positive HC2 or cytology peaked in the age group 35-39 years and was lowest in the age group 55-60 years. High-grade squamous intraepithelial lesion or worse lesions (HSIL+) were histologically detected in 23.5% of women with positive baseline cytology and in 9.8% of women with positive baseline HC2 only on follow-up. All women with histologic HSIL+ had positive baseline HC2. The hrHPV test is superior to cytology in the early detection of high-grade cervical epithelial lesions. In this study, the prevalence of histologic HSIL+ on follow-up of women with positive hrHPV test was rather high, and these women should be kept under careful surveillance. In northern Thailand, hrHPV testing has a potential to be used as a primary screening test for cervical cancer with cytology applied as a triage test.

Cervical cancer screening by high risk HPV testing in routine practice: results at one year recall of high risk HPV-positive and cytology-negative women

Cervical cancer screening by human papillomavirus (HPV) testing requires the use of additional triage and follow-up analyses. We evaluated women's compliance with and the performance of this strategy in a routine setting. Five cervical service screening programmes in North-East Italy. Eligible women aged 25-64 invited for a new screening episode underwent HPV testing for high risk types (hrHPV by Hybrid Capture 2) and cytology triage. Women with positive HPV and cytology results were referred for colposcopy; women with positive HPV but negative cytology results were referred to 1-year repeat hrHPV testing. Of 46,694 women screened by HPV testing up to December 2011, 3,211 (6.9%) tested hrHPV positive; 45% of these had a positive triage cytology. Those with negative cytology were invited for 1-yr repeat testing. Compliance with invitation was 61.6% at baseline and 85.3% at 1-yr repeat. Rate of persistent hrHPV positivity was 58% (830/1,435). Colposcopy performed in women with a positive hrHPV test at 1-yr repeat accounted for 36% of all colposcopies performed within the screening programmes. Cumulatively, a histological high-grade lesion was detected in 276 women (5.9‰ detection rate), 234 at baseline (85%), and 42 (15%) at 1-yr repeat. Compliance with hrHPV-based screening programmes was high both at baseline and at 1-yr repeat. Compared with the randomized trials, a higher proportion of triage cytology was read as positive, and only a small number of high-grade lesions were detected among the group of hrHPV positive cytology negative women who repeated testing 1-yr after baseline.

Hr-HPV testing in the follow-up of women with cytological abnormalities and negative colposcopy

Background:The follow-up after abnormal Pap smear and negative colposcopy is not clearly defined. This study aimed at investigating the role of hr-HPV testing in the management of abnormal Pap test and negative colposcopy for Cervical Intraepithelial Neoplasia grade 2 or worse (CIN2+).Methods:The study enroled 1029 women with abnormal screening cytology (years 2006–2010) and negative colposcopy for CIN2+, which subsequently performed a hr-HPV test. Incident CIN2+ lesions were identified through linkage with cancer registry, hospital discharge records, neoplastic pathology reports and the archive of screening programme (2006–2011).Results:During the follow-up, the cohort developed 133 CIN2+ lesions; only one among hr-HPV-negative women. The probability of developing CIN2+ on follow-up time was 0.44% (95% confidence interval (CI) 0.1–3.1) and 41.8% (95% CI 31.8–53.5) for hr-HPV-negative women and hr-HPV-positive women, respectively. A woman with a positive hr-HPV test had about 105 times higher probability of developing a CIN2+ lesion than a woman with a negative hr-HPV test (hazard ratio (HR)=104.5, 95% CI 14.5–755.1), adjusted for index Pap test result, age and cervix squamocolumnar junction visualisation.Conclusion:Our results confirm that hr-HPV testing is able to select the real group of women at risk of developing CIN2+ lesions in the follow-up of abnormal cytology and first negative colposcopy.

HPV DNA testing in population-based cervical screening (VUSA-Screen study): results and implications.

Background:Human papillomavirus (HPV) testing is more sensitive than cytology for detecting high-grade cervical intraepithelial neoplasia (CIN). We evaluated the performance of high-risk HPV (hrHPV) testing in routine screening.Methods:In all, 25 871 women (29–61) enrolled in our population-based cohort study were offered both cytology and hrHPV testing. High-risk HPV-positive women with normal cytology and an age-matched subcohort of hrHPV-negative women with normal cytology were invited for repeat testing after 1 and/or 2 years and were referred for colposcopy if they presented with abnormal cytology and/or a positive hrHPV test. The hrHPV-positive women with borderline or mild dyskaryosis (BMD) and all women with moderate dyskaryosis or worse (>BMD) were directly referred for colposcopy. Women with BMD and an hrHPV-negative test were advised to repeat cytology at 6 and 18 months and were referred for colposcopy if the repeat cytology test was abnormal. The main outcome measure was CIN grade 3 or worse (CIN3+). Results were adjusted for non-attendance at repeat testing.Results:The hrHPV-positive women with abnormal cytology had a CIN3+ risk of 42.2% (95% confidence interval (CI): 36.4–48.2), whereas the hrHPV-positive women with normal cytology had a much lower risk of 5.22% (95% CI: 3.72–7.91). In hrHPV-positive women with normal cytology, an additional cytology step after 1 year reduced the CIN3+ risk to only 1.6% (95% CI: 0.6–4.9) if the repeat test was normal. The CIN3+ risk in women with hrHPV-positive normal cytology was higher among women invited for the first time (29–33 years of age) (9.1% 95% CI: 5.6–14.3) than among older women (3.0% 95% CI: 1.5–5.5).Conclusion:Primary hrHPV screening with cytology triage in women aged ⩾30 years is an effective way to stratify women on CIN3+ risk and seems a feasible alternative to cytological screening. Repeat cytology after 1 year for hrHPV-positive women with normal cytology is however necessary before returning women to routine screening.

Factors predicting the outcome of conservatively treated adenocarcinoma in situ of the uterine cervix: An analysis of 166 cases

ObjectiveThe present study assessed the clinical outcome of patients conservatively treated for cervical adenocarcinoma in situ (AIS) and their predictive factors using univariate and multivariate population averaged (PA) generalized estimating equation (GEE) model in a longitudinal setting. MethodsA series of 166 consecutive women (mean age 39.8yrs; range 23–63yrs) underwent conservative treatment of AIS as the primary treatment and were followed-up (mean 40.9mo) using colposcopy, PAP-smear, biopsy and HPV-testing with Hybrid Capture 2. ResultsHysterectomy was performed as part of the primary management in 47 patients, who were excluded from the follow-up (FU) analysis. Out of 119 women closely followed-up, additional therapeutic procedures were performed in 69. At study conclusion, 7 patients (5.9%) showed persistent disease, while 8 (6.7%) had progressed to invasive adenocarcinoma (AC). Positive HR-HPV test was the only independent predictor of disease recurrence (adjusted OR=2.72; 95%CI 1.08–6.87), and together with free cone margins (OR=0.20; 95%CI 0.04–0.92), HR-HPV positivity was also the single most powerful predictor of disease progression to AC, with OR=3.74; 95%CI 1.84–7.61 (p=0.0001) in multivariate PA-GEE. ConclusionsThese results suggest that testing HR-HPV positive at any time point during FU is the most significant independent predictor of progressive disease, while showing free margins in cone has a significant protective effect against progression to AC. Furthermore, because 4.3% women with persistent, recurrent or progressive disease experienced a late (5th and 6th FU) diagnosis of HG-CGIN or microinvasive AC, a close surveillance should be scheduled for at least three years in conservatively treated AIS patients.

Correlation between hybrid capture II high‐risk human papillomavirus DNA test chemiluminescence intensity from cervical samples with follow‐up histologic results

The Hybrid Capture II high-risk human papillomavirus (hrHPV) DNA test is a US Food and Drug Administration-approved nucleic acid hybridization assay using chemiluminescence for the semiquantitative detection of hrHPV in cervical samples. Patient samples and controls are used to calculate results as negative for hrHPV if <1.0, positive for hrHPV if >2.5, and "equivocal" if between 1.0 and 2.5. The authors reported on the cervical histologic results of 209 patients demonstrating "equivocal" results for hrHPV from SurePath (204 patients) or ThinPrep (5 patients) vials, and compared patients in this cohort with atypical squamous cells of undetermined significance (ASC-US) cytology on the index cervical Papanicolaou (Pap) test (Group 1; n = 148 patients) with a patient cohort demonstrating unequivocal positive hrHPV test results (Group 2; n = 148 patients). The chemiluminescence intensity of hrHPV tests from patients in Group 2 were correlated with the presence and severity of dysplasia on subsequent histologic results, and patients were thereby stratified for their subsequent risk of cervical intraepithelial neoplasia (CIN) types II/III. Approximately 97% of hrHPV tests demonstrating "equivocal" results were found to be positive at the time of retesting, and 15% of biopsied cases demonstrated CIN II or III. Results of follow-up histology after an ASC-US diagnosis, expressed as a percentage of the biopsied cohort, were: CIN II/III: 16.5% in Group 1 and 22.4% in Group 2; CIN I: 27% in Group 1 and 23.5% in Group 2; and negative: 56.5% in Group 1 and 54.1% in Group 2. Chemiluminescence intensity did not appear to be correlated with the severity of dysplasia. The percentage of high-grade CIN in the "equivocal" hrHPV cohort is highly significant and therefore the management of these patients should be similar to the unequivocally positive population. After an unequivocal positive hrHPV test, the hrHPV chemiluminescence intensity does not appear to further predict the rate of high-grade CIN.

Significance of High-Risk Human Papillomavirus DNA Detection in Women 50 Years and Older With Squamous Cell Papanicolaou Test Abnormalities

Data on cytologic screening and follow-up disproportionately reflect findings from frequently screened younger women, and data from screened women 50 years and older using newer screening technologies remain limited. To better understand the utility of adjunctive high-risk human papillomavirus (hrHPV) testing for disease risk stratification in women 50 years and older with a range of liquid-based cytology, abnormal, squamous cell Papanicolaou test results. Liquid-based cytology cases interpreted as high-grade squamous intraepithelial lesion (HSIL); low-grade squamous intraepithelial lesion (LSIL); atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H); and atypical squamous cells of undetermined significance (ASC-US) at Magee-Womens Hospital (Pittsburgh, Pennsylvania) were retrospectively identified for a 36-month period between July 1, 2005, and June 30, 2008, from women 50 years and older who also had hrHPV DNA test results. Histopathologic follow-up diagnoses were analyzed. During the study period, 4855 women 50 years and older had HSIL, LSIL, ASC-H, or ASC-US Papanicolaou test results and hrHPV testing. In 89.3% of HSIL cases, 71.0% of LSIL cases, 38.9% of ASC-H cases, and 14.2% of ASC-US cases, hrHPV test results were positive. The positive predictive value of a positive hrHPV test for histopathologic cervical intraepithelial neoplasia 2/3(+) detection was 90.5% with HSIL, 15% with ASC-H, 9.8% with LSIL, and 3.2% with ASC-US. A negative hrHPV test result had a 100% negative predictive value for histopathologic cervical intraepithelial neoplasia 2/3(+) in both LSIL and ASC-H cases. In women 50 years and older, a positive hrHPV test result significantly increased the likelihood of follow-up histopathologic diagnoses of cervical intraepithelial neoplasia 2 /3(+) in patients with HSIL, LSIL, and ASC-H Papanicolaou test results compared with women with negative hrHPV test results. No cervical intraepithelial neoplasia 2/3(+) diagnoses were documented in women 50 years and older with LSIL or ASC-H Papanicolaou test results and negative hrHPV test results.

Colposcopic Characteristics of High-Risk Human Papillomavirus-Related Cervical Lesions

Because high-risk human papillomavirus (hrHPV) is the necessary factor in the development of high-grade cervical lesions, knowing the colposcopic differences between hrHPV-positive and hrHPV-negative lesions could be of value. We have evaluated whether there are colposcopic differences between lesions testing hrHPV-positive or hrHPV-negative. This study was conducted as a retrospective case-control study. We designed a scoring system for colposcopic criteria that might be of relevance to distinguish hrHPV-positive from hrHPV-negative lesions. Colposcopic images were analyzed from patients at the VU University Medical Center Amsterdam, the Netherlands, in whom a GP5+/6+ polymerase chain reaction hrHPV test had been performed within a month of the colposcopic examination (n = 507). Visibility of the transformation zone (corrected for age), a (very) coarse and irregular punctation pattern, and a large lesion (>25% of the visible cervix) were more often associated with a positive hrHPV test status (p =.001, odds ratio [OR] = 2.29, 95% CI = 1.41-3.73; p =.036, OR 2.37, 95% CI = 1.08-5.19; and p =.044, OR = 1.78, 95% CI = 1.08-2.94, respectively). After correction for histologic diagnosis, the difference between hrHPV-positive and hrHPV-negative lesions for visibility of the transformation zone and lesion size remained statistically significant (OR = 2.44, 95% CI = 1.35-4.41 and OR = 1.92, 95% CI = 1.04-3.54, respectively). This study indicates that visibility of the transformation zone, (very) coarse punctation pattern, and larger lesion size were the main colposcopic features associated with a hrHPV-positive test status.

Validation of high-risk HPV tests for primary cervical screening

From a clinical point of view, testing for a broad spectrum of high-risk human papillomavirus (hrHPV) is only useful when a positive hrHPV test result is informative about the presence of high-grade cervical intraepithelial neoplasia or cervical cancer (CIN 2 or worse). Two hrHPV tests, i.e. HC2 and GP5+/6+, have shown in large clinical trials that they perform better in the detection of CIN 2+/CIN 3+ lesions than cytology and thus have been clinically validated. Consequently these tests are now considered as alternative screening tools for cytology in cervical screening. Candidate hrHPV tests to be used for cervical screening should have a similar balance between sensitivity and specificity for CIN 2+ lesions as these two clinically validated hrHPV tests in order to prevent redundant or excessive follow-up procedures for women with transient hrHPV infections or hrHPV-positive women without cervical lesions. The data from these large prospective clinical studies can be used to set standards for the clinical performance and characteristics of the candidate hrHPV test. To prevent costly validation trajects of candidate hrHPV tests and based on the available data from large clinical studies we demonstrate how guidelines for hrHPV test requirements and guidelines for clinical validation of candidate hrHPV tests have been developed, and how these guidelines should be used in cervical screening. It is expected that the use of these guidelines will facilitate implementation of hrHPV testing in primary cervical screening.

Contribution of DNA ploidy image cytometry to the management of ASC cervical lesions

The Bethesda system classifies smears that suggest an underlying cervical intraepithelial neoplasia (CIN) as ASC (atypical squamous cell) smears. ASC smears are subdivided into ASCUS (of undetermined significance) and ASCH (cannot exclude a high-grade lesion). Today the management of ASCUS is a triage with HR-HPV testing and colposcopy is recommended for ASCH. The aim was to conduct a study on ASC smears to determine DNA ploidy measurement for the detection of CIN2+. The link between a suspect DNA ploidy assessed by image cytometry and/or a positive HR-HPV testing was analyzed on 69 ASCUS and 82 ASCH smears, and the presence of CIN2+ within 12 months after ASC diagnosis. The ploidy was suspect in case of aneuploidy, multiploidy, or in the presence of cells with a DNA content >5c or >9c. Every woman who had a CIN2+ had a suspect DNA profile in the ASCUS smears and every woman except 1 was HR-HPV-positive. The link between a positive HR-HPV test or a suspect DNA profile or both and a CIN2+ was high (P = .019, .023, and .008, respectively). The presence of >9c cells was particularly linked to CIN2+ (P = .0031). In all, 90.9% and 87.9% of the ASCH smears with CIN2+ were, respectively, HR-HPV positive or had a suspect ploidy (P = .0000 and P = .0043), and the presence of >9c cells was also linked to CIN2+ (P = .003). HR-HPV testing and determination of the ploidy profile with special attention to 9c-exceeding cells could be accurate for a better management of ASC smears.

Alternating high-risk human papillomavirus infection

Nearly every Dutch woman will be exposed to genital human papillomavirus (HPV) at least once during her lifetime, and most likely several times. In the current study, the authors investigated the prevalence of high-risk-HPV (HR-HPV) infection and the likelihood of progression to cervical intraepithelial neoplasia (CIN). In this study, the course of HR-HPV infection in 703 women was observed. From a database of 720,016 negative cytology smears, the authors selected 703 women based on the availability of at least 2 HR-HPV polymerase chain reaction tests. The authors database stores not only the HPV data but also all other cytologic and histologic data, allowing the detection of women who progressed from negative cytology to CIN within a period of 10 years. Of the 703 selected women, 159 were found to have alternating HR-HPV infection (change from a negative HR-HPV test to a positive test or vice versa), 40 had a persistently positive HR-HPV test, and 504 women had a persistently negative HR-HPV test. The percentage of alternating HPV infection declined over time from 37% to 7%. Of the women age older than 40 years, 17% had an alternating HR-HPV infection, 2 of whom developed CIN. These findings led the authors to conclude that all the women in the current study with an increased risk of developing type 2 or 3 CIN were identified using 2 HPV tests. Women age older than 40 years still have a significant risk of acquiring a HR-HPV. In light of the current study findings, the authors believe it is worth considering the inclusion of women age 40 years and older who have negative cytology for HPV testing as part of the Dutch national screening program.

Routine cervical screening with primary HPV testing and cytology triage protocol in a randomised setting

The role of high-risk human papillomavirus (hrHPV) testing in primary cervical screening has not been established. We generated a randomised evaluation design ultimately to clarify whether primary hrHPV testing implemented into routine screening can bring increase in the programme effectiveness. The aim of the present report on first-year results was to assess the cross-sectional relative validity parameters for routine hrHPV screening, in comparison with conventional screening. An equal number of women invited to routine screening was randomly allocated to primary hrHPV screening (n=7060) and to cytological screening (n=7089). In the hrHPV screening arm, after a single positive hrHPV test result, the need of colposcopy referral was determined by a cytological triage test. Compared with the conventional arm, more colposcopy referrals were made in the hrHPV screening arm (relative risk 1.51, confidence interval 95% 1.03–2.22). Specificity of the primary screening with sole hrHPV test (91.5–92.1%) was much lower than that with the cytology triage (98.7–99.3%), which was not quite as specific as screening with conventional cytology (99.2–99.6%). Compared with conventional cytology, primary screening with hrHPV test results in increased cross-sectional relative sensitivity at the level of all positive lesions at the cost of substantial loss in specificity. With cytology triage, the specificity improves to the level of conventional cytology.

POBASCAM, a population‐based randomized controlled trial for implementation of high‐risk HPV testing in cervical screening: Design, methods and baseline data of 44,102 women

Cytological cervical screening is rather inefficient because of relatively high proportions of false negative and false positive smears. To evaluate the efficiency of high-risk human papillomavirus (hrHPV) testing, by GP5+/6+ PCR-enzyme immunoassay (EIA), in conjunction with cytology (Intervention Group) to that of the classical cytology (Control Group), we initiated the Population Based Screening Study Amsterdam (POBASCAM). POBASCAM is a population-based randomized controlled trial for implementation of hrHPV testing in cervical screening. The outcome measure is the proportion of histologically confirmed > or =CIN3 lesions in each study arm up to and including the next screening round after 5 years. We present the design, methods and baseline data of POBASCAM. When, in the next 5 years, the follow-up will be completed, the data obtained will be used in model studies, including a cost-effectiveness study, to advise the Dutch Ministry of Public Health in deciding whether cervical screening should be based on combined hrHPV and cytology testing instead of cytology alone. Between January 1999 and September 2002, 44,102 women (mean age = 42.8 years; range = 29-61) that participated in the regular Dutch screening program were included in our study. In the Intervention Group the distribution of cytology and hrHPV by cytology class was as follows: normal cytology 96.6% (3.6% hrHPV positive); borderline and mild dyskaryosis (BMD) 2.5% (34.6% hrHPV positive); and moderate dyskaryosis or worse (>BMD) 0.8% (88.3% hrHPV positive), i.e., 0.4% moderate dyskaryosis (82.9% hrHPV positive), 0.3% severe dyskaryosis (92.5% hrHPV positive), 0.1% carcinoma in situ (95.2% hrHPV positive), <0.1% suspected for invasive cancer (hrHPV positive 100.0%). In the Control Group 96.5% of the women had normal cytology, 2.4% BMD and 0.8% >BMD, i.e., 0.4% moderate dyskaryosis, 0.3% severe dyskaryosis, 0.1% carcinoma in situ, <0.1% suspected for invasive cancer. The presence of hrHPV was age-dependent, decreasing from 12.0% at 29-33 years to 2.4% at 59-61 years. Among women with a positive hrHPV test, the prevalence of BMD was age-dependent ranging from 20.2% at 29-33 years to 7.8% at 54-58 years. In contrast, the risk of >BMD of 13.7% among women with a positive hrHPV test was not age-dependent. Our study indicates that large-scale hrHPV testing by GP5+/6+ PCR-EIA in the setting of population-based cervical screening is practically feasible, is accepted by both participating women and general practitioners and yields highly reproducible results.

Actualisation de la conduite à tenir en cas de lésions à papillomavirus humains (HPV)

Human papillomaviruses (HPV) are the principal cause of cervical carcinomas. It is now well accepted that HPV testing is particularly useful in the triage of ASCUS. HPV testing can also be used as a primary screening tool. In our experience using the Hybrid Capture II® assay in 10 569 women, the sensitivity of HPV testing for detecting a histologically proven HGSIL was 100 % and 97,5 %, higher than that of conventional cytology (72.7 %) and liquid-based (80,3 %). The low specificity of 87 % of HPV testing was slightly increased to 89,5 % if HPV testing was reserved to women > 30 years old. The quantitative approach provided by the HC-II assay for the assessment of the viral load could not be used for predicting a HGSIL. The negative predictive value of the HPV testing was close to 100 %. HR-HPV testing could be proposed in primary screening of HGSIL in association with cytology. It significantly improves the detection of HGSIL with conventional cytology. A positive HR-HPV test selects population of women at high risk for developing HGSIL. Considering the high specificity of liquid-based cytology in our series (94,8 %), cytology on the same sample could be only performed in this population. In another way, a negative HR-HPV test selects populations at low risk who could benefit a different algorithm of screening of 2 to 3 years.