Equivocal Cytology Research Articles

Performance of fluorescence in situ hybridization in biliary brushings with equivocal cytology: an institutional experience

Biliary brushing (BB) cytology has a sensitivity of 15%-65% and specificity approaching 100% for detecting malignancy. Fluorescence in-situ hybridization (FISH) using the UroVysion probe set has been advocated to enhance the detection of malignancies with reported sensitivity of 43%-84%. We sought to evaluate the performance of FISH in BB with equivocal cytology at our institution. Patients with atypical and suspicious BB with concurrent diagnostic FISH performed at our institution from 2014 to 2021 were identified through a query of our pathology database. FISH (using UroVysion probe set containing centromere enumeration probes to chromosomes 3, 7, and 17) was positive if at least 5 cells demonstrated polysomy. Electronic medical records were reviewed for pathology results and outcomes. Patients were classified malignant if they had positive pathology or documented clinical impression of malignancy and benign if they had negative pathology and/or documented benign clinical course for at least 12 months. We identified 254 equivocal BB (238 atypical/16 suspicious) with concurrent FISH results from 191 patients (105 benign, 86 malignant). 12% (22/191) of patients were FISH positive. Twenty-four percent (21/86) of patients with malignancy had positive FISH but were nonspecific for pancreaticobiliary/ampullary adenocarcinomas. Almost all positive FISH were associated with malignancy (21/22; 95%). There was 1 positive FISH in a patient with primary sclerosing cholangitis who had a benign outcome. The small number of positive FISH results in BB with equivocal cytology raises the question of the optimal criteria for malignancy. Using only polysomy could result in lower sensitivity.

A single tertiary institution review of the international system for serous fluid cytopathology and the impact of cell block and ancillary studies on its performance.

We sought to assess the utility of the International System for Serous Fluid Cytopathology (TIS) in the context of our department's routine practice. We examined 1028 archived effusion cytology (pleural, peritoneal, and pericardial) cases from 2018 to 2019, and re-classified them along the international system into the following diagnostic categories: nondiagnostic (ND), negative for malignancy (NFM), atypia cells of undetermined significance (AUS), suspicious for malignancy (SFM), and malignant (MAL). The full distribution of the cases examined was as follows: ND 2.0%; NFM 66.1%; AUS 6.0%; SFM 4.7%; MAL 21.2%. Overall risk of malignancy for each category was calculated as: ND 30.0%; NFM 18.0%; AUS 61.9%; SFM 100%; MAL 94.4%. The overall performance attributes of TIS were as follows: sensitivity 57.1%; specificity 98.3%; positive predictive value 94.4%; negative predictive value 82.0%; diagnostic accuracy 84.5%. The new classification was simple and intuitive to use and our results appear to fall within the expected ranges of the new guidelines, with risk of malignancy and accuracy comparable to similar studies. The availability of a cell block allowed for refinement of the diagnosis in a majority of cases with equivocal cytology, though this was dependent on the cell yield.

The utility of claudin-4 versus MOC-31 and Ber-EP4 in the diagnosis of metastatic carcinoma in cytology specimens.

Claudin-4 is a sensitive and specific marker for carcinoma in effusion cytology. The authors examined the diagnostic use of claudin-4 versus MOC-31 and Ber-EP4 by comparing their sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in differentiating carcinoma from mesothelioma and benign/mesothelial hyperplasia in effusion specimens. This retrospective study comprised a cohort of 229 cytology specimens, including 211 effusion fluid and 18 fine-needle aspiration specimens. Cytologic categories included 134 carcinoma, 28 mesothelioma, 46 indefinite (suspicious and atypical), and 21 benign. Cell block sections were stained for claudin-4 and compared with those previously stained for MOC-31 and Ber-EP4. Indefinite cases were further reclassified based on clinical and pathologic findings into benign (26 cases), mesothelioma (11 cases), and carcinoma (nine cases). None of the mesotheliomas (0/39) or benign effusions (0/47) were positive for claudin-4, whereas 134 of the 143 carcinoma specimens were positive. Compared to MOC-31 and Ber-EP4, claudin-4 had the highest specificity and PPV (100% for each), followed by Ber-EP4. Claudin-4 showed high sensitivity (93.7%), albeit lower than MOC-31. MOC-31 had the lowest specificity and PPV but the highest sensitivity and NPV. Ber-EP4 had the lowest sensitivity (91.6%). Claudin-4 can be used as a single marker for carcinoma with high sensitivity and superior specificity compared with MOC-31 and Ber-EP4. Mesothelial lineage can be ruled out when claudin-4 is positive. In equivocal cytology samples with few scattered cells of interest, a panel of claudin-4 and Ber-EP4 results in the highest combined sensitivity and specificity.

Combined Shear Wave Elastography and EU TIRADS in Differentiating Malignant and Benign Thyroid Nodules.

Although multimodal ultrasound approaches have been suggested to potentially improve the diagnosis of thyroid cancer; the diagnostic utility of the combination of SWE and malignancy-risk stratification systems remains vague due to the lack of standardized criteria. The purpose of the study was to assess the diagnostic value of the combination of grey scale ultrasound assessment using EU TIRADS and shear wave elastography. 121 patients (126 nodules-81 benign; 45 malignant) underwent grey scale ultrasound and SWE imaging of nodules between 0.5 cm and 5 cm prior to biopsy and/or surgery. Nodules were analyzed based on size stratifications: <1 cm (n = 43); 1-2 cm (n = 52) and >2 cm (n = 31) and equivocal cytology status (n = 52), and diagnostic performance assessments were conducted. The combination of EU TIRADS with SWE using the SD parameter; maintained a high sensitivity and significantly improved the specificity of sole EU TIRADS for nodules 1-2 cm (SEN: 72.2% vs. 88.9%, p > 0.05; SPEC: 76.5% vs. 55.9%, p < 0.01) and >2 cm (SEN: 71.4% vs. 85.7%, p > 0.05; SPEC: 95.8% vs. 62.5%, p < 0.01). For cytologically-equivocal nodules; the combination with the SWE minimum parameter resulted in a significant reduction in sensitivity with increased specificity (SEN: 60% vs. 80%; SPEC: 83.4% vs. 37.8%; all p < 0.05). SWE in combination with EU TIRADS is diagnostically efficient in discriminating nodules > 1 cm but is not ideal for discriminating cytologically-equivocal nodules.

The prognostic value of urinary cytology after trimodal therapy (TMT) for muscle-invasive bladder cancer.

Urine cytology and cystoscopy are routinely employed during follow-up of patients after trimodal therapy (TMT) for muscle-invasive bladder cancer (MIBC). The significance of positive or equivocal cytology without visible disease recurrence on cystoscopy during follow-up is unknown, and studies informing outcomes in this scenario are lacking. This study aims to investigate the temporal trends of positive/equivocal cytology in the absence of visible disease recurrence and the association with bladder cancer recurrence and survival outcomes. One hundred and twenty-nine patients with available post-TMT cytology data and negative cystoscopy from a single academic institution between 2002 and 2017 with a median follow-up of 3.4 (range 0.1-14.2) years were analyzed. Cytology results, first post-TMT cytology positive/equivocal (CP) and negative (CN), were evaluated for association with disease recurrence and survival. Kaplan. Meier and competing risks methods were used to assess time-to-negative cytology in CP patients with ≥2 interval post-TMT cytology results (n = 33), time-to-recurrence, and disease-specific mortality (DSM) stratified by first post-TMT cytology result. At first follow-up (6-8 weeks post-TMT completion), CP was observed in 41 (32%) and CN in 88 (68%) of patients. With further follow-up of CP patients with ≥2 interval post-TMT cytology results, the probability of developing negative cytology was 57% (95% CI 42, 77) at 6 months post-TMT, and the median time-to-negative cytology was 3.2 months (95% CI 2.99, 5.80). The median time-to-recurrence was reduced in CP patients compared to CN (24.3 vs. 78.1 months, p = 0.1), corresponding with an apparent increase in the cumulative incidence of recurrence rate at 3 years in the CP vs. CN group (62% vs. 42%, p = 0.1). No significant difference was observed in the 3-year DSM rates. On univariable analysis, the hazards of recurrence and DSM for patients with CP were 1.5 (95% CI 0.9, 2.5, p = 0.1) and 2.1 (95% CI 0.9, 4.7, p = 0.07) respectively. This is the first study to investigate the significance of a positive/equivocal cytology without visible disease following TMT for MIBC. Positive cytology is common and does not preclude subsequent negative cytology supporting a watchful waiting approach rather than proceeding immediately to biopsy. However, cytology that remains positive at subsequent follow-up may be associated with adverse recurrence and survival outcomes.

Role of colposcopy in the management of women with abnormal cytology.

Cytology identifies the women who are at higher risk of harboring high-grade cervical premalignant lesions or invasive cancer. However, a diagnostic test such as colposcopy is crucial for women with abnormal cytology for localization of the abnormality, confirmation of diagnosis, and appropriate management. To standardize this subjective technique and to minimize the interobserver variations, Swede scoring system was introduced. The revised colposcopic nomenclature of the International Federation of Cervical Pathology and Colposcopy in 2011 included various normal and abnormal colposcopic findings and gives a description of colposcopic features which improves its accuracy over the colposcopic indices. There is consensus agreement that cytology indicative of high-grade lesions (ASC-H and HSIL in the Bethesda system) should engender immediate referral for colposcopy and biopsy. The management of women who have equivocal or borderline cytology of low-grade abnormalities (ASCUS/LSIL) is still under deliberation. It is generally agreed to have an HPV triage for women with equivocal cytology. Based on the latest recommendations, the current chapter provides an extensive overview of the role of colposcopy in the management of women with various abnormalities reported on Pap smear.

Low reproducibility of equivocal categories of the Bethesda System for Reporting Thyroid Cytology makes the associated risk of malignancy specific to the diagnostic center

PurposeEquivocal categories (III, IV, V) of the Bethesda System for Reporting Thyroid Cytology (BSRTC) are characterized by high variability of the estimated risk of malignancy. The aim of the study was to analyze the reproducibility of classification of nodules into an equivocal category and the frequency of malignancy (FoM) observed in such categories.MethodsFive experienced cytopathologists from three centers (A, B, C) independently performed reclassification of smears obtained from 213 thyroid nodules with equivocal routine cytology and known results of the postoperative histopathological examination.ResultsThe interobserver agreement among all cytopathologists was poor, with a Krippendorff’s alpha coefficient equaling 0.34. The intra-center agreement was higher than the inter-center (fair vs poor). Pathologists of the center A classified smears into categories II and III significantly less often and categories IV and V more often than pathologists of centers B and C. The joint FoM of nodules classified into any of categories IV–VI (regarded as an indication for surgery) was different among centers (A: 40.0%, B: 66.7%, C: 80.6%). The FoM of category III nodules with features of nuclear atypia (AUS) in center B and C was two times higher than that of other nodules of category III (FLUS), while in center A the FoM was similar.ConclusionsThe use of published data on the risk of malignancy in nodules of particular BSRTC categories without concern for the uniqueness of the diagnostic center may lead to erroneous conclusions.

Primary HPV screening for cervical cancer: Results after two screening rounds in a regional screening program in Finland.

Since 2012, cervical cancer screening has been conducted with a primary high-risk human papillomavirus (hrHPV) test and conventional cytology triage in the city of Tampere, Finland. The women who were screened with the hrHPV test in 2012 were invited to participate in the second screening round in 2017. The aim of the present report was to compare the number of colposcopy referrals and the number of histological high-grade squamous intraepithelial lesion (HSIL)+ (cervical intraepithelial neoplasia [CIN2+]) lesions between the first and second screening rounds of women of a specific age group who were screened twice with the hrHPV test. The primary hrHPV test used was the RealTime hrHPV PCR assay by Abbott. Women with a positive hrHPV test and cytology triage equal to or worse than low-grade squamous intraepithelial lesion or atypical glandular cells, favor neoplasia, were directly referred to colposcopy, whereas hrHPV-positive women with a negative or equivocal cytology triage were re-screened after approximately 12-16months. hrHPV-negative women were scheduled for re-screening after 5years. The present report focuses on the cohort of women who were screened twice with the hrHPV test, who were 35-55years old in 2012, and 40-60years old in 2017. In all, 8076 women were invited for HPV screening in 2012 and 8331 women were invited for the second round 5years later, with attendance rates of 70% and 71%, respectively. Of the women who were screened in 2012, 4571 (69%) belonged to the 35- to 55-year age cohort. In 2017, 4807 (73%) of the women aged 40-60years participated in the screening. In this cohort, 185 (4.0%) colposcopies were performed in the first screening round, compared with 139 (2.9%) in the second round, and the colposcopy rate was 29% smaller in the second round (P=.002). The number of histological HSIL+ cases was 38 (0.8%) during the first screening round and 29 (0.6%) during the second round (P=.220). In the setting of routine organized cervical cancer screening, the initially high colposcopy rate associated with primary HPV screening seems to level off at the second screening round in women who were screened twice with an hrHPV test.

Usability of EU-TIRADS in the Diagnostics of Hürthle Cell Thyroid Nodules with Equivocal Cytology.

The aim of this study was to compare the diagnostic effectiveness of EU-TIRADS in two groups of nodules with equivocal cytology (categories III-V of Bethesda system), with and without Hürthle cells (HC and non-HC). The study included 162 HC and 378 non-HC nodules with determined histopathological diagnosis (17.9% and 15.6% cancers). In both groups calculated and expected risk of malignancy (RoM) for high, intermediate and benign risk categories of EU-TIRADS were concordant. RoM for low risk category was higher than expected in both groups, but especially in HC (HC: 13.9%, non-HC: 7.0%, expected: 2–4%). The majority of cancers in HC of that category were follicular thyroid carcinomas (FTC) and Hürthle cell thyroid carcinoma (HTC) (60.0% vs. non-HC: 16.7%). The diagnostic efficacy of EU-TIRADS was lower in HC (the area under the receiver operating characteristics curve (AUC): 0.621, sensitivity (SEN): 44.8%, specificity (SPC): 78.9% for high risk threshold) than in non-HC (AUC: 0.711, SEN: 61.0%, SPC: 77.7%). AUC was the highest for category V (AUC > 0.8, both groups) and the lowest for category IV (inefficient, both group). If intermediate risk category was interpreted as an indication for surgery, 25% of cancers from category III and 21.4% from category IV would not be treated in the HC group (0.0% and 7.4% from non-HC group, respectively). EU-TIRADS does not aid making clinical decisions in patients with cytologically equivocal HC nodules, particularly those classified into category IV of Bethesda System for Reporting Thyroid Cytopathology (BSRTC).

Performance of novel non-invasive urine assay UroSEEK in cohorts of equivocal urine cytology.

Urine cytology is an essential element of the diagnostic work up of hematuria. A significant proportion of cases continue to be placed in the "atypical" or "suspicious" categories of the Paris system for urine cytology, posing difficulty in patient management. We report on the performance of our recently described urine-based assay "UroSEEK" in cases with equivocal diagnosis in patients who are investigated for bladder cancer. Urine samples were collected from two cohorts. The first consisted of patients who presented with hematuria or lower urinary tract symptoms (early detection cohort) and the second of patients that are in follow-up for prior bladder cancer (surveillance cohort). Urine samples were analyzed for mutations in 11 genes and aneuploidy. In the early detection setting, we found high sensitivity and specificity (96% and 88%, respectively) and a strong negative predictive value of 99%. The assay performance was less robust in the surveillance cohort (sensitivity of 74%, specificity of 72%, and negative predictive value of 53%). UroSEEK demonstrated a notable lead time to cancer diagnosis. Seven cases in the early detection cohort and 71 surveillance cases were detected at least 6months prior to clinical diagnosis. Our results suggest a potential role for UroSEEK assay in guiding management of patients with atypical urine cytology if confirmed in future prospective trials.

Diagnostic performance of a stepwise cytological algorithm for biliary malignancy in primary sclerosing cholangitis.

Detection of early cholangiocarcinoma (CCA) in primary sclerosing cholangitis (PSC) is challenging. The aim of this study was to evaluate the diagnostic accuracy of a stepwise approach to biliary brush cytology with sequential use of fluorescence in-situ hybridization (FISH) for the detection of biliary malignancy in PSC. We retrospectively studied consecutive patients with PSC who underwent biliary brushings at Karolinska University Hospital between 2009 and 2015 (n=208). Brush samples were categorized as benign, equivocal (atypical or suspicious) and malignant. Equivocal cases were further analysed with FISH. Samples with a malignant cytology or positive FISH were considered positive. The diagnosis was determined after 12months of follow-up. The diagnosis CCA was confirmed in 15 patients (7%), high-grade dysplasia in three patients, and low-grade dysplasia in five patients at follow-up. Using the diagnostic algorithm, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) for a diagnosis of CCA were 80% (95%CI 52%-96%), 96% (95%CI 92%-98%), 60% (95%CI 36%-81%) and 98% (95% CI 95%-100%). In patients with equivocal cytology (n=61), the sensitivity for CCA diagnosis increased to 100% (95%CI 72%-100%) with a lower PPV of 58% (95%CI 34%-78%). The diagnostic accuracy for detection of CCA in all patients was 95% (95%CI 91%-97%). Biliary brush cytology with sequential use of FISH in equivocal cases seems to be a highly predictive diagnostic test for CCA in PSC. These results support the use of FISH when cytology is equivocal for detection of biliary malignancy in PSC.

Clinical Utility of CINtec PLUS Triage in Equivocal Cervical Cytology and Human Papillomavirus Primary Screening.

While cervical cytology is accepted triage for human papillomavirus (HPV)-positive women, the efficiency of cervical screening could be improved by exploiting disease markers with higher specificity. CINtec PLUS triage alone and combined with HPV 16/18 genotyping was performed on ThinPrep samples from HPV-positive women. Clinical performance and the potential to reduce or expedite colposcopy referrals were evaluated. The 2-year sensitivity and specificity for cervical intraepithelial neoplasia grade 3 or worse (CIN3+) were 90% and 42%, respectively. Specificity was improved over HPV testing in equivocal cytology and could cut the referral rate by about 40%. When combined with HPV 16/18 genotyping, CINtec PLUS triage of the 12 other high-risk HPV genotypes generally demonstrated better sensitivity for CIN3+ than separate triage of non-type-specific HPV-positive women. This strategy could reduce colposcopy referrals by 31%. These findings highlight the potential of CINtec PLUS to improve management pathways in HPV-positive women. CINtec PLUS cytology represents a sensitive and efficient triage in HPV-positive women. The clinical performance of the dual-stain was most notable in women younger than 25 years and could potentially improve management pathways.

Editorial Comment

The appeal of urinary biomarkers is an obvious one. The potential to diagnose a disease earlier, direct treatment more efficiently, and assess for progression or recurrence could all be game-changing tools in our armamentarium. Indeed, PCA3 has already been FDA-approved for prostate cancer assessment and the AUA/SUO Joint Guideline on non-muscle invasive bladder cancer established an Expert Opinion that UroVysion® FISH and ImmunoCyt™ may be used to assess response to intravesical BCG and adjudicate equivocal cytology. 1 Chang S.S. Boorjian S.A. Chou R. et al. Diagnosis and treatment of non-muscle invasive bladder cancer: AUA/SUO guideline. http://www.auanet.org/education/guidelines/non-muscle-invasive-bladder-cancer.cfmDate: 2016 Google Scholar Other biomarkers for both types of cancers are well into development and testing.

Urinary NID2 and TWIST1 methylation to augment conventional urine cytology for the detection of bladder cancer

Abnormal methylation of urinary TWIST1 and NID2 conferred high sensitivity and specificity for the detection of urothelial carcinoma. We examine the performance of the urine-based TWIST1/NID2 methylation assay with the addition of urine cytology for the detection of urothelial carcinoma. A prospective multi-institutional study was conducted to assess the performance of a methylation assay for patients with hematuria or under surveillance for non-muscle invasive bladder cancer (NMIBC). All patients underwent cystoscopy, a methylation assay, and cytology. Receiver operator characteristic (ROC) curves were constructed for cytology alone, the methylation assay alone, and a combined model. Areas under the curve (AUC) were compared using likelihood ratio tests. A total of 172 patients were enrolled (37% for hematuria and 63% NMIBC). The AUC for cytology alone with equivocal cytologies positive was 0.704, and improved to 0.773 with the addition of the DNA methylation assay (p < 0.001). When the equivocal cytologies were considered negative, the AUC improved from 0.558 to 0.697 with the addition of the DNA methylation assay (p = 0.003). Addition of a TWIST1/NID2-based DNA methylation assay adds diagnostic value to urine cytology and the model is sensitive to the classification of equivocal cytology.

Thyroid cancer and nodular goiter of thyroid: An analysis of patients in rural South Tamil Nadu

Background: The aim of the study was to analyze the profile of nodular goiters of thyroid presenting with thyroid cancers in a tertiary hospital in rural South India. Materials and Methods: This study was conducted for 2 years since 2010 in patients with thyroid enlargements. Patients with multinodular or solitary thyroid enlargements with ultrasonogram suspicious of malignancy, fine-needle aspirate suspicious of malignancy, or persons with clinical suspicious thyroid enlargements with equivocal ultrasonogram or cytology with or without symptoms of mass effect were selected for the study. The clinical, operative, cytological, and histological data were tabulated and statistically analyzed. Results: Of 522 patients analyzed, 91.57% were females and 67.62% were between 21 and 40 years. Nearly 17.04% presented with hypothyroidism, 15.13% with hyperthyroidism, and 67.82% patients were euthyroid. Of the 522 patients, 425 (81.42%) had nodular thyroid enlargements, of whom 224 (42.91%) had multinodular goiters (MNGs), 201 (38.51%) had solitary thyroid nodules (SNT), and 97 (18.58%) had diffuse thyroid enlargements. The malignant tumors were present in 105 patients with papillary carcinoma, 20 patients with follicular carcinoma, and 6 with medullary carcinoma, of whom 76 (33.93%) had MNG and 54 (26.87%) had SNT. Conclusion: Thyroid cancers are best diagnosed through concerted clinical, radiological, cytological, and histological analyses. All nodules of thyroid, whether solitary or multinodular, should be sampled in fine-needle aspiration under radiological assistance and similarly all nodules need to be studied for malignancy in histopathology. In this study, we observed that the incidence rates of malignancy were higher than that of the existing Indian data and these patients were from the coastal regions of Tirunelveli and Thoothukudi districts. We also observed that multinodular goiters had microscopic malignant foci.

Diagnostic yield of cytopathology in evaluating pericardial effusions: Clinicopathologic analysis of 419 specimens.

Pericardial effusions can cause considerable morbidity and potentially may lead to mortality. Malignant pericardial effusions are uncommon, and data on malignancies encountered in pericardial effusion cytology specimens are limited. Relevant records of all pericardial effusions from January 2008 to September 2014 were examined and compared with pericardial biopsy results when performed. Discrepant cases were reviewed to determine the cause of the disagreement. In total, 419 pericardial effusion specimens obtained from 364 patients were examined. Cytologic diagnostic categories included: negative for malignancy (332 specimens; 79%), equivocal (25 specimens; 6%), and positive (62 specimens from 51 patients; 15%). Forty-seven patients who had positive effusions were known to have malignancy. The most common primary malignancies were breast (39.3%) and lung (39.3%) cancers in women and lung cancer (47.4%) in men. A concurrent pericardial biopsy was performed in 46% of patients. Excluding equivocal cytologic diagnoses, cytology and biopsy were concordant in 153 of 173 paired samples (88.4%). The sensitivity of cytology in diagnosing malignancy was 92.1% compared with 55.3% for pericardial biopsy. Cytologic examination has significant diagnostic utility in the evaluation of pericardial effusions and exhibits a lower false-negative rate compared with pericardial biopsy. Submission of pericardial biopsy alongside effusion cytology is associated with increased sensitivity for detecting malignancy and may be especially useful in the setting of low-volume pericardial effusion. Cancer Cytopathol 2017;125:128-137. © 2016 American Cancer Society.

Relevance of HPV Screening for Triaging Equivocal Cytology Findings in the Pap II-p, Pap III and Pap IIID Groups - Results of Two Long-Term Studies.

Introduction: The use of HPV screening for the triage of ASC-US (atypical squamous cells of undetermined significance) cytology results has been established as a sound standard by international trials whereas the data for other cytology findings are in part contradictory. There is a lack of long-term studies on the use of HPV triage in Germany. Materials and Methods: For the present study data from a primary HPV screening project involving women aged over 30 years, ongoing since 2006, and an epidemiological study on women aged between 20 and 27 years, ongoing since 2009, were used. Upon recruitment, all women underwent a smear test for cytology and screening for "high-risk" HPV using Hybrid Capture 2 (HC2). If both tests were positive or if there were persisting remarkable cytology findings or a positive HPV test, then clarification by colposcopy was performed. Results: Altogether, among 282 women with Pap II-p (ASC-US), Pap III (ASC-H) or Pap IIID (LSIL + CIN2) and negative HPV test there was no case of CIN3+. Among the women under 30 years of age, however, 69 % (ASC-US) to 85 % (LSIL + CIN2) of the remarkable findings were HPV positive, also among the older women with Pap IIID, the 71 % prevalence of HPV was too high for a triage and even without triage there was a 23 % risk for CIN3+. On the other hand, of the women over 30 years old with ASC-US (Pap II-p) findings, only 21 % were positive for HPV and the risk for CIN3+ in this group was high at 29 %. Also for ASC-H (Pap III) findings in the age group of over 30 years with an HPV prevalence of 56 % there was an efficient triage for CIN3+. Discussion: In summary, the HPV triage of ASC-US (Pap II-p) findings in women aged over 30 years was found to be efficient; in contrast, LSIL + CIN2 (Pap IIID) findings in this age group justified an immediate referral to colposcopy whereas cytology control appeared to be sufficient for younger women.

Correlation of Cobas HPV Testing and Follow-Up Biopsies in Women with Negative Pap Tests

High risk HPV (hrHPV) testing is commonly used as co-testing in cervical cancer screening or as a reflex test in cases with equivocal cytology. Recently the FDA approved the Cobas HPV test as a screening option in women above 25 years. Clinical data regarding the use of Cobas HPV testing in women with negative Pap tests is limited. Utilizing a large series of cases, this study analyzed the correlation of hrHPV tests and histologic biopsies in women with negative Pap tests. INTRODUCTION RESULTS MATERIALS AND METHODS

A study of HPV typing for the management of HPV-positive ASC-US cervical cytologic results

BackgroundIn US cervical screening, immediate colposcopy is recommended for women with HPV-positive ASC-US (equivocal) cytology. We evaluated whether partial typing by Onclarity™ (BD) might identify HPV-positive women with low enough CIN3+ risk to permit 1-year follow-up instead. MethodsThe NCI-Kaiser Permanente Northern California Persistence and Progression cohort includes a subset of 13,890 women aged 21+ with HC2 (Qiagen)-positive ASC-US at enrollment; current median follow-up is 3.0years. Using stratified random sampling, we typed 2079 archived enrollment specimens including 329 women subsequently diagnosed with CIN3+, 563 with CIN2, and 1187 with <CIN2. Adjusting for sampling, we computed 3-year cumulative CIN3+ risks for each Onclarity typing channel, using Kaplan–Meier methods. ResultsThe 3-year CIN3+ risk for all HC2-positive women with ASC-US was 5.2%; this establishes the “benchmark” risk for colposcopic referral. Hierarchically, 3-year cumulative risks for each typing channel were 16.0% for HPV16, 7.4% for HPV18, 7.0% for HPV31, 7.1% for grouped HPV33/58, 4.3% for HPV52, 3.9% for HPV45, 2.7% for HPV51, 1.6% for HPV39/68/35, and 1.3% for HPV59/56/66. DiscussionASC-US linked to HPV16, HPV18, HPV31, or HPV33/58 warrants immediate colposcopy. Optimal management of women with HPV52 or HPV45 is uncertain. Risk of women with only HPV51, HPV39/68/35, or HPV59/56/66 might be low enough to recommend 1-year retesting permitting viral clearance. This strategy would defer colposcopy for 40% of women with HPV-positive ASC-US, half of whom would be cotest-negative at 1-year return. Approximately 10% of those with CIN3 diagnosable at enrollment would be delayed 1year instead. Cost-effectiveness analyses are needed.

Comparison of HPV genotyping and methylated ZNF582 as triage for women with equivocal liquid-based cytology results.

IntroductionThe interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). Recently, methylated zinc finger protein 582 (ZNF582) (ZNF582m) was reported to be highly associated with cervical cancer. In this study, we compared the performance of ZNF582m detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H).Case descriptionTwo hundred and forty-two subjects with equivocal papanicolaou smear (Pap smear) results were recruited in this hospital-based and case-controlled study. The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and then for ZNF582m and HPV-HR detection. The level of ZNF582m was quantified by real-time methylation-specific PCR after bisulfite conversion. The HPV-HR test was performed by using a nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and HPV type-specific primers.Discussion and evaluationSignificant associations were observed between ZNF582m and the risk of cervical intraepithelial neoplasia grade 3 or higher (CIN3+; odds ratio = 15.52, 95% confidence interval (CI): 7.73 to 31.18). The sensitivity and specificity of ZNF582m for women with CIN3+ were 82.43% and 76.79%, respectively. High sensitivity (99.33%) but low specificity (38.76%) was observed for HPV-HR. When combining both positive results of ZNF582m and HPV-HR, the sensitivity and specificity were 82.43% and 81.55%, respectively. The sensitivity and specificity of ZNF582m or HPV-16/18 were 89.19% and 70.24%, respectively. However, the sensitivity and specificity of ZNF582m combined with HPV-16/18 (both ZNF582m and HPV-16/18 positive results) were 59.46% and 94.64%, respectively.ConclusionsZNF582m provides a promising triage tool for women with ASC.To effectively manage ASC patients, a new strategy co-testing for ZNF582m and HPV-16/18 genotyping was proposed. This strategy could reduce the number of patients referred for colposcopic examination and thus provide a feasible follow-up solution in the regions where colposcopy is not readily available. This strategy could also prevent women from experiencing unnecessary anxiety caused by HPV-HR.