Diagnosis Of Atypical Squamous Cells Of Undetermined Significance Research Articles

Is it Really Necessary to Perform Colposcopy in Patients with Ascus and HR HPV Positivity?

IS IT REALLY NECESSARY TO PERFORM COLPOSCOPY IN PATIENTS WITH ASCUS AND HR HPV POSITIVITY? INTRODUCTION: Papanicolaou (Pap) smear is a very important screening method for detection of cervİcal cancer or cell changes that can lead to it. Atypical squamous cells of undetermined significance (ASCUS) is the most common abnormality seen among all cervical cytologies with the percentage of 4-5% . Human papillomavirus (HPV) infection is regarded as a major cause of cervical intraepithelial neoplasia (CIN) and cervical cancer. Colposcopy is recommended for all women with high-grade squamous intraepithelial lesion (HSIL) and atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion (ASC-H) for cervical cancer screening . It is also recommended for high-risk HPV (HR HPV) positive women with ASCUS. The aim of this study is to evaluate the importance of colposcopy in patients with ASCUS and HR HPV positivity. MATERIALS AND METHODS: Study Population We performed a retrospective study approved by the ethics committee of Acıbadem University. Women with diagnosis of ASCUS and HR HPV positivity who underwent colposcopic evaluation at 2 different gynecologic oncology clinics of our university from January 2011 to January 2019 were included in our study. The age range was from 21 to 48 and the mean age of patients’ was 29. Subjects who were pregnant, hysterectomized or had previous cytological abnormalities were excluded. CONCLUSION: In this study, we aimed to evaluate the significance of colposcopy in patients with ASCUS and high-risk HPV positivity, especially strains 16 and 18. As mentioned in some other studies, immediate colposcopy is an expensive screening procedure for further evaluation of ASCUS .

Comparison between Urine and Cervical High-Risk HPV Tests for Japanese Women with ASC-US.

Most uterine cervical cancers are caused by the persistent infection of the high-risk human papillomavirus (hrHPV). Thus, the hrHPV-DNA test, which examines specimens from the cervix, is the standard screening method as well as cytology in western countries. Urine sampling for the hrHPV-DNA test would be easier and help improving screening rates. This study prospectively investigated the concordance between urine and cervical hrHPV tests for patients with atypical squamous cells of undetermined significance (ASC-US) in cervical cytology. We recruited 338 women with the cytologic diagnosis of ASC-US and performed hrHPV-DNA tests to both samples from the uterine cervix and first void urine, using the Cobas 4800 system. In all hrHPV genotypes, the simple concordance rate was 90.8% (307/338) and the Kappa statistic value was 0.765, which shows substantial concordance. The positive concordance rate was 70.5% (74/105), which was the rate excluding women who had negative results in both tests. When limited to types 16 and 18, the simple concordance rate was 98.8% (334/338), and the Kappa statistical value was calculated to be 0.840, which showed almost perfect concordance. The positive concordance rate resulted in 81.8% (18/22). We conclude that the urine hrHPV-DNA test could substitute the cervical test in women with ASC-US.

CytoProcessorTM: A New Cervical Cancer Screening System for Remote Diagnosis

Background: Current automated cervical cytology screening systems still heavily depend on manipulation of glass slides. We developed a new system called CytoProcessor^TM (DATEXIM, Caen, France), which increases sensitivity and takes advantage of virtual slide technology to simplify the workflow and save worker time. We used an approach based on artificial intelligence to identify abnormal cells among the tens of thousands in a cervical preparation. Objectives: We set out to compare the diagnostic sensitivity and specificity of CytoProcessor^TM and the ThinPrep Imaging System (HOLOGIC, Marlborough, MA, USA). Methods: A representative population of 1,352 cases was selected from the routine workflow in a private laboratory. Diagnoses were established using the ThinPrep Imaging System and CytoProcessor^TM. All discordances were resolved by a consensus committee. Results: Compared to the ThinPrep Imaging System, CytoProcessor^TM significantly improves diagnostic sensitivity without compromising specificity. The sensitivity of detection of “atypical squamous cells of undetermined significance (ASC-US) and more severe” and “low-grade squamous intraepithelial lesion and more severe” was significantly higher using CytoProcessor^TM. Considering that cases with a truth diagnosis of ASC-US or more severe required clinical follow-up, 1.5% of the cases (21/1,360) would have been missed if the CytoProcessor^TM diagnosis had been used for clinical decision-making. In contrast, 4% of the cases (54/1,360) were missed when the ThinPrep Imaging System diagnosis was used for clinical decision-making. There were 2.6 times fewer false negatives using CytoProcessor^TM. The CytoProcessor^TM workflow was 1.5 times faster in terms of worker time. Conclusions: CytoProcessor^TM is the first of a new generation of automated screening systems, demonstrating improved sensitivity and yielding significant gains in processing time. In addition, the fully digital nature of slide presentation in CytoProcessor^TM allows the remote diagnosis of Papanicolaou tests for the first time.

Increased Rate of ASCUS Diagnosis With Concomitant Request for High-Risk Human Papillomavirus Reflex Testing May Be Due to Cognitive Bias

To determine if concomitant high-risk human papillomavirus (HPV-HR) reflex testing may bias the cytologic interpretation of Papanicolaou (Pap) tests. Percentage of atypical squamous cells of undetermined significance (ASCUS) and HPV-HR positivity was compared between Pap tests with HPV-HR cotesting and HPV-HR reflex testing for ASCUS, with subset analysis of cytopathologists' experience. ASCUS in the reflex group (41.5%) was significantly higher than the cotesting group (33.0%) (P = .02). There was no difference in HPV-HR positivity or ASCUS/squamous intraepithelial lesion (SIL) ratios between the two groups. The cytopathologists' experience inversely correlated with the proportion of ASCUS but did not explain the higher reflex group ASCUS. HPV-HR reflex testing may introduce bias in cytologic diagnosis, making it more likely that an ASCUS diagnosis is rendered. HPV-HR and ASCUS/SIL ratios were similar between the groups, so cytopathologist performance was not significantly affected. There was no effect of cytopathologists' experience.

Risks of cervical intraepithelial neoplasia grade 3 or invasive cancers in ASCUS women with different management: a population-based cohort study.

ObjectiveTo investigate the progression risk of atypical squamous cells of undetermined significance (ASCUS) with different clinical managements.MethodsWomen with their first diagnosis of ASCUS cytology were retrieved from the national cervical cancer screening database and linked to the national health insurance research database to identify the management of these women. The incidences of developing cervical intraepithelial neoplasia grade 3 and invasive cervical cancer (CIN3+) were calculated, and the hazard ratios (HRs) were estimated using a Cox proportional hazards model. This study was approved by the Research Ethics Committee of the National Taiwan University Hospital and is registered at ClinicalTrials.gov (Identifier: NCT02063152).ResultsThere were total 69,741 women included. Various management strategies including colposcopy, cervical biopsies and/or endocervical curettage, and cryotherapy, failed to reduce the risk of subsequent CIN3+ compared with repeat cervical smears. Loop electrosurgical excision procedure/conization significantly decreased risk of subsequent CIN3+ lesions (HR=0.22; 95% confidence interval [CI]=0.07–0.68; p=0.010). Women in their 40s–50s had an approximately 30% risk reduction compared to other age groups. Women with a previous screening history >5 years from the present ASCUS diagnosis were at increased risk for CIN3+ (HR=1.24; 95% CI=1.03–1.49; p=0.020).ConclusionIn women of first-time ASCUS cytology, a program of repeat cytology can be an acceptable clinical option in low-resource settings. Caution should be taken especially in women with remote cervical screening history more than 5 years.

Cumulative High-Grade Squamous Intraepithelial Lesion Rate and Need for Surgical Intervention of Atypical Squamous Cells of Undetermined Significance Cytology-Diagnosed Patients: A Prospective Study

Objective: Τhe study aimed to study the rate of atypical squamous cells of undetermined significance (ASCUS) that progressed to high-grade squamous intraepithelial lesions (HGSIL), as well as the number of patients who finally necessitated a surgical intervention during follow-up. Methods: A prospective study was conducted on patients admitted for colposcopy during 2007-2012. We exclusively included those who presented with newly ASCUS diagnosis, while patients with a history of cervical intraepithelial neoplasia (CIN) were excluded. Primary end points were the cumulative rate of HGSIL during follow-up and the rate of surgical procedures performed because of such lesions. Results: There were 134 ASCUS cases included. Overall, there were 48 (35.8%) surgical excision procedures performed to treat or eliminate HGSIL during the follow-up period. According to the final histopathology of surgical specimens, the cumulative rate of CIN2 or higher cervical lesions was 28.4% (n = 38). Conclusion: The cumulative rate of CIN2+ during follow-up period was almost 30% for patients with ASCUS, with the necessity for interventional treatment being even higher.

Seguimiento de pacientes con atipia de células escamosas de significado indeterminado (ASCUS) en un servicio citológico de Medellín (Colombia), 2011-2015

Introducción: en la literatura científica nacional existen pocos estudios sobre el seguimiento de mujeres con atipias de células escamosas de significado indeterminado (ASCUS). Objetivo: describir el seguimiento a cinco años de una cohorte retrospectiva de mujeres con resultado de ASCUS en un servicio citológico de Medellín, Colombia. Materiales y métodos: se realizó un estudio de una cohorte retrospectiva de mujeres con ASCUS en una Institución Prestadora de Servicios de Salud de Medellín, entre 2011 y 2015. Se evaluaron aspectos socio-demográficos, signos, síntomas y hallazgos preneoplásicos a partir de una fuente de información secundaria. Se determinó la frecuencia de mujeres sin control citológico, con regresión de la lesión, persistencia o progresión a lesión intraepitelial escamosa (LIE). Se estimó la proporción de incidencia de ASCUS y LIE en cada año de seguimiento. Resultados: se incluyeron 2.771 mujeres con diagnóstico de ASCUS. La incidencia de regresión al primer control fue 69,8% y en el último 84,6% y de progresión 10,8% y 6,6%, respectivamente, con menos del 1,0% de progresión a LIE de alto grado. La incidencia de ASCUS osciló entre 3,6% (2012) y 5,7% (2013) mientras que de LIE entre 1,1% (2012) y 2,4% (2014). El 28,1% de la población nunca asistió a un control. Conclusiones: se halló una elevada frecuencia de regresión de las lesiones clasificadas como ASCUS y de mujeres sin control durante cinco años; esto evidencia la necesidad de mejorar las estrategias de seguimiento citológico que redunden en la disminución de costos asociados a la implementación de otras pruebas diagnósticas.

Associations between sexually transmitted infections, high-risk human papillomavirus infection, and abnormal cervical Pap smear results in OB/GYN outpatients.

ObjectiveThis study aimed to examine the meaning and usefulness of sexually transmitted infection (STI) test when caring for patients who have abnormal cervical cytology and/or positive high-risk human papillomavirus (HPV) DNA test results.MethodsAmong patients who underwent liquid-based cervical cytology and HPV DNA tests at the Obstetrics and Gynecology outpatient clinic, 800 patients who showed abnormal cervical cytology were compared with 200 patients in the control group. Both groups were simultaneously tested via multiplex real-time polymerase chain reaction for seven types of STI-causative microorganisms.ResultsThe positive rate of high-risk HPV infection in total STIs positive group was 1.47 times higher than that of total STIs negative group. The probability of a cytological diagnosis of a grade equal to or higher than atypical squamous cells-cannot exclude high grade squamous intraepithelial lesion (ASC-H) was significantly higher in patients testing positive for total STIs (1.46 times), Chlamydia trachomatis (3.21 times), or Mycoplasma genitalicum (3.58 times) than in those testing negative. The total STIs positive rate was significantly higher for those having a cytological diagnosis of a grade equal to or higher than atypical squamous cells of undetermined significance (ASC-US) when high-risk HPV test result was negative.ConclusionCorrelations were present not only between STIs and high-risk HPV infection but also between abnormal cervical cytology and STIs. Therefore, additional evaluation of STIs will be helpful to appropriately diagnose and treat patients with abnormal cervical cytology, positive results on high-risk HPV DNA test, or a cytological diagnosis of ASC-US despite negative high-risk HPV DNA test result.

Improving the performance of reflex Human Papilloma Virus (HPV) testing in triaging women with atypical squamous cells of undetermined significance (ASCUS): A restrospective study in a tertiary hospital in United Arab Emirates (UAE)

IntroductionCervical cancer is the second commonest cancer in women worldwide. Infection with oncogenic types of human Papillomavirus (HPV) is the most important risk factor for developing cervical cancer. Reflex High risk HPV (HR-HPV) testing is of significant value in the assessment of Papanicolaou (Pap) smear results where ASCUS are identified. ObjectiveTo improve the performance of reflex HR-HPV testing in triage of ASCUS and analyze the factors impacting it. MethodologyIn this study, we generated a database of 9641 women who had cervical smears collected during the study period from the cytopathology record in a large tertiary hospital in UAE. These included 297 smears with ASCUS diagnosis. All cases were retrospectively followed up with a mean duration of 2.44 years. We analyzed data according to the outcome based on several follow-up Pap smear analysis as the reference assessment. ResultsWe detected HR-HPV infection in 17.9% of cases. 9.1% <25, 28.8% 25–34 and 62.1% ≥35 years old. HR-HPV prevalence was higher among premenopausal women (20.7%) compared to postmenopausal women (9.5%) (P-value=0.044). The rate of progression to high grade lesions was also higher (28.7%) in the premenopausal group compared to (12.8%) in the postmenopausal group. Reflex HPV testing had an overall sensitivity of 41.1%, specificity of 88.2%, positive predictive value (PPV) of 62.1%, and negative predictive value (NPV) of 75.9% in detection of cervical lesions. These figures were higher on combining premenopausal status and complaint of abnormal bleeding or discharge/itching (66.7%, 93.3%, 66.8% and 93.3% respectively). ConclusionsThe sensitivity, specificity and NPV of reflex HPV testing in the triage of ASCUS cases can be more accurate in premenopausal women upon adding age group and presenting complaint as a triage item. This improves the performance of reflex HPV testing and the subsequent selection of high risk patients for colposcopy.

The utility of high-risk HPV testing in the management of women 30 to 64 years of age with low-grade squamous intraepithelial lesions on cervical cytology

The current American Society for Colposcopy and Cervical Pathology guidelines recommend high risk-human papillomavirus testing for patients 30 to 64 years of age with a low-grade squamous intraepithelial lesion on Papanicolaou (Pap) tests. This study presents our experience since the introduction of the new guidelines. From September 1, 2013 through August 31, 2014, 20,014 ThinPrep Pap tests were processed, of which 17,989 (90%) were negative, 863 (4.3%) were atypical squamous cells of undetermined significance (ASCUS), and 540 (2.7%) were low-grade squamous intraepithelial lesions (LSIL). Thirty-eight percent of the negative cases, 87% of the ASCUS cases, and 76% of the LSIL cases were processed for high risk-human papillomavirus genotypes. Of the 1,163 patients with a Pap test diagnosis of ASCUS and LSIL, 656 (56%) underwent colposcopic examination and, of those, 240 (37%) had cervical biopsies performed. In our cohort, 247 (33%) of the 754 ASCUS Pap tests were hr-HPV-positive and 507 (67%) were hr-HPV-negative. Of the 409 LSIL Pap tests, 245 (60%) were hr-HPV-positive and 164 (40%) were hr-HPV-negative. Colposcopy/biopsy follow-up within 3-months of the ThinPrep Pap test showed cervical intraepithelial lesion grade 2/3 lesions in 16% of the hr-HPV-positive ASCUS cases, in 15% of the hr-HPV-positive LSIL cases and in 9% of the hr-HPV-negative LSIL cases. This study supports the current management guidelines of colposcopy for women 30 to 64 years of age with hr-HPV-positive ASCUS and LSIL on Pap tests and repeat Pap test with cotesting for women with hr-HPV-negative LSIL.

Clinical significance of ASCUS and ASC-H cytological abnormalities: a six-year experience at a single center.

To evaluate colposcopic biopsy results of patients with cervical cytological findings of atypical squamous cells of undetermined significance (ASC-US) and atypical squamous cells with high-grade lesions that cannot be excluded (ASC-H). A retrospective evaluation of data from 358 patients, who had cervical cytological findings of ASC-US (n = 335) and ASC-H (n = 23), and had colposcopic assessments between 2005 and 2011. Cervical biopsy results of patients diagnosed with ASC-US cytology (n = 335) revealed cervical squamous cell carcinoma 0.9 % (n = 3) at biopsy, cervical intraepithelial neoplasia 3 (CIN 3) in 3.8 % (n = 13), cervical intraepithelial neoplasia 2 (CIN 2) in 1.1 % (n = 4), cervical intraepithelial neoplasia 1 (CIN 1) in 35.2% (n = 118), and benign lesions in 59 % (n = 197). Cervical biopsy results of patients diagnosed with ASC-H cytology (n = 23) revealed CIN 3 at biopsy in 39.3% (n = 9), CIN 2 in 21.7% (n = 5), CIN 1 in 26% (n = 6), carcinoma in situ in 8.7% (n = 2), and squamous cell cancer in one patient (4.3%). The cytological diagnosis of ASC-US may lead to the diagnosis of cervical intraepithelial lesion of higher grades as well as cervical cancer and should be evaluated by colposcopic cervical biopsy.

Cervical Cytology and Human Papillomavirus Testing in Adolescent Women: Implications in Management of a Positive HPV Test.

Objectives. Consensus guidelines establish that HPV testing should not be used to manage adolescents with atypical squamous cells of undetermined significance (ASC-US). This study aimed to estimate the impact on follow-up of HPV testing after the first-time ASC-US diagnosis. Methods. From January 2009 to December 2010, all ASC-US diagnoses in adolescents were retrieved. Results. 1950 cervical cytologies were received from this population and 335 cases (17.1%) were reported as ASC-US. A total of 287 cases were included in the study. Cases were divided into control group (no HPV test; 46 cases) and case group (HPV test performed; 241 cases). On follow-up, in the control group, 43.4% patients had cytology, and 56.6% patients had no follow-up. The case group was divided into negative HPV (60 cases) and positive HPV (181 cases). In the negative-HPV group, 41.7% had cytology and 58.3% had no follow-up. In the positive-HPV group, 41% had cytology, 22% underwent colposcopy, and 37% had no follow-up. Patients with positive-HPV results were more likely to have follow-up than patients in the control and negative-HPV groups (63% versus 43.4% versus 41.7%, resp.). Conclusions. HPV infections are common in adolescents. A positive HPV test cannot predict which women will develop carcinoma. Adherence to current guidelines is recommended in this population.

Testing of Integrated Human Papillomavirus mRNA Decreases Colposcopy Referrals: Could a Change in Human Papillomavirus Detection Methodology Lead to More Cost-Effective Patient Care?

Objectives: This study investigates potential colposcopy referral rates, as per the latest American Society for Colposcopy and Cervical Pathology recommendations, following the change in high-risk human papillomavirus (HR-HPV) detection methodology from Hybrid Capture 2 (HC2) to APTIMA at our institution. Study Design: Rates of colposcopy referral were compared between two cohorts, each comprising all Pap samples with a diagnosis of atypical squamous cells of undetermined significance (ASCUS) tested for HR-HPV in our laboratory during a 12-month period. Cohorts I and II included Pap samples tested with HC2 (n = 1,856) and APTIMA (n = 1,651), respectively. The rates of quantity not sufficient (QNS) results were determined for all Pap samples during the same time periods. Results: The proportion of HR-HPV-positive Pap samples with an ASCUS diagnosis was significantly lower with APTIMA (42%) than with HC2 (53%; p < 0.0001). APTIMA also resulted in a significantly lower QNS rate among all Pap samples (0.42 vs. 4.3% with HC2; p < 0.0001). Conclusion: The change in HR-HPV detection methodology from HC2 to APTIMA has led to a 21% reduction in colposcopy referrals and a 90% decrease in QNS rates at our institution. The new methodology has resulted in more cost-effective patient care and fewer insufficient samples requiring repeat HR-HPV testing.

Reply to triage of women with minor abnormal cervical cytology: Meta‐analysis of the accuracy of an assay targeting messenger ribonucleic acid of 5 high‐risk human papillomavirus types

BACKGROUND High-risk human papillomavirus (hrHPV) DNA detection is generally accepted for the triage of women with a cytologic diagnosis of atypical squamous cells of undetermined significance (ASC-US). However, no consensus has been reached on the optimal management of low-grade squamous intraepithelial lesions (LSIL). METHODS In this meta-analysis, the diagnostic accuracy of nucleic acid sequence-based amplification (NASBA) detection of messenger ribonucleic acid (mRNA) of 5 hrHPV types (the PreTect HPV-Proofer and NucliSENS EasyQ tests) for detecting grade 2 cervical intraepithelial neoplasia or worse (CIN2+) and CIN3+ was assessed in women who had a diagnosis of ASC-US and LSIL. The results were compared with the Hybrid Capture-2 (HC2) assay, which detects the DNA of 13 hrHPV types. A bivariate random-effect model that incorporated the intrinsic correlation between the true-positive and false-positive rates was used for a pooled meta-analysis. RESULTS Considering underlying CIN2+, the pooled absolute sensitivity of the 10 included studies was 75.4% (95% confidence interval [CI], 68.1%-82.7%) and 76.2% (95% CI, 68.3%-76.9%) for the triage of ASC-US and LSIL, respectively. The pooled absolute specificity to exclude CIN2+ was 77.9% (95% CI, 70.1%-85.7%) and 74.2% (95% CI, 69.5%-78.8%) in women with ASC-US and LSIL, respectively. Five studies allowed direct comparison of the mRNA assays with HC2. Considering CIN2+ in women with ASC-US and LSIL, mRNA testing was substantially more specific than the HC2 assay (ratio: 1.98 and 3.36, respectively; P < .001) but was less sensitive (ratio: 0.80 and 0.74, respectively; P < .001). CONCLUSIONS HPV assays for detecting the mRNA of 5 hrHPV types may reduce the over-diagnosis of women who have minor cytologic abnormalities. However, given the lower sensitivity, women with negative mRNA test results cannot be considered free of CIN2+ and require further surveillance. Cancer (Cancer Cytopathol) 2013;121:675–687. © 2013 American Cancer Society.

Increased rate of atypical squamous cells of undetermined significance and declining high-risk human papillomavirus rates following implementation of ThinPrep Imaging System are associated with increased nuclear chromasia

The ThinPrep Imaging System ([Imager], Cytyc Corporation, Hologic, Inc.) for cervical cytology is commonly employed in laboratories. It uses a proprietary stain that allows measurement of cellular DNA content. We evaluated the initial effect of implementing the Imager stain on atypical squamous cells of undetermined significance (ASCUS) and squamous intraepithelial lesion (SIL) diagnoses, and high-risk human papillomavirus (hrHPV) rates. This study included 264 Imager-stained ASCUS cases from the first 5 months of Imager use. All cases were analyzed for hrHPV using Hybrid Capture II assay (Digene Corporation, Qiagen). ASCUS/SIL ratios and hrHPV-positive rates were calculated for the laboratory overall and for 3 cytopathologists. ASCUS/SIL ratios were also calculated for 5 cytotechnologists. These metrics were compared with those from the equivalent 5-month period 1 year prior. Slide review of all Imager-stained ASCUS cases was performed to assess for potential factors involved in ASCUS misinterpretation. The proportion of ASCUS diagnoses increased after Imager stain introduction, from 1.79% to 3.14% (P < 0.001). The proportion of SIL diagnoses also increased, from 0.69% to 1.1% (P < 0.001). However, the hrHPV-positive rate declined from 61.3% to 53.6% (P = 0.104). Review of Imager-stained ASCUS cases showed marked nuclear hyperchromasia, particularly in benign squamous metaplastic cells. Imager stain implementation, required for the ThinPrep Imaging System, led to significant increase in ASCUS rates while the hrHPV-positive rate declined. Cytopathologists will need to recalibrate thresholds for an ASCUS diagnosis when interpreting Imager-stained slides. In particular, nuclear hyperchromasia in squamous metaplastic cells should not be overdiagnosed as ASCUS.

Triage of women with minor abnormal cervical cytology: Meta‐analysis of the accuracy of an assay targeting messenger ribonucleic acid of 5 high‐risk human papillomavirus types

High-risk human papillomavirus (hrHPV) DNA detection is generally accepted for the triage of women with a cytologic diagnosis of atypical squamous cells of undetermined significance (ASC-US). However, no consensus has been reached on the optimal management of low-grade squamous intraepithelial lesions (LSIL). In this meta-analysis, the diagnostic accuracy of nucleic acid sequence-based amplification (NASBA) detection of messenger ribonucleic acid (mRNA) of 5 hrHPV types (the PreTect HPV-Proofer and NucliSENS EasyQ tests) for detecting grade 2 cervical intraepithelial neoplasia or worse (CIN2+) and CIN3+ was assessed in women who had a diagnosis of ASC-US and LSIL. The results were compared with the Hybrid Capture-2 (HC2) assay, which detects the DNA of 13 hrHPV types. A bivariate random-effect model that incorporated the intrinsic correlation between the true-positive and false-positive rates was used for a pooled meta-analysis. Considering underlying CIN2+, the pooled absolute sensitivity of the 10 included studies was 75.4% (95% confidence interval [CI], 68.1%-82.7%) and 76.2% (95% CI, 68.3%-76.9%) for the triage of ASC-US and LSIL, respectively. The pooled absolute specificity to exclude CIN2+ was 77.9% (95% CI, 70.1%-85.7%) and 74.2% (95% CI, 69.5%-78.8%) in women with ASC-US and LSIL, respectively. Five studies allowed direct comparison of the mRNA assays with HC2. Considering CIN2+ in women with ASC-US and LSIL, mRNA testing was substantially more specific than the HC2 assay (ratio: 1.98 and 3.36, respectively; P < .001) but was less sensitive (ratio: 0.80 and 0.74, respectively; P < .001). HPV assays for detecting the mRNA of 5 hrHPV types may reduce the over-diagnosis of women who have minor cytologic abnormalities. However, given the lower sensitivity, women with negative mRNA test results cannot be considered free of CIN2+ and require further surveillance.

Verification of doubtful PAP smear results of women included in the screening program in the Podlaskie province

Verification of uncertain PAP-smear results in a group of women covered by the cervical screening program in the Podlaski province. The main aim of the study was to identify CIN (Cervical Intraepithelial Neoplasia) lesions present, with varying degrees of severity in women with cytological diagnosis of ASCUS (atypical squamous cells of undetermined significance), LSIL (low grade squamous intraepithelial lesion), and ASC-H (atypical squamous cells - cannot exclude high grade squamous intraepithelial lesion). The study evaluated 101 cervical smears taken from the vaginal part of the cervix in a group of screened women in the Podlaski province. Cytological evaluation was performed according the Bethesda System. We analyzed abnormal smears selected from a total of 7296 cytological examinations performed during 2012 at the University Center for Pathomorphological and Genetic - Molecular Diagnosis, Medical University in Białystok. The cytological results which were of interest to us included 19 cases with ASCUS, 59 with LSIL, and 23 with ASC-H, as well as with morphological features of the presence of Human Papilloma Virus (HPV). Staining was performed using CINtecPLUS test according to the manufacturer's instructions. CINtecPLUS is a immunocytochemical test based on specially designed monoclonal antibodies (E6H4TM) that let us identify protein p16ink4a within the cervical smear Additionally the diagnostic kit was provided with antibodies for diagnosing the presence of Ki-67 protein, a known marker of cell proliferation. The result was considered positive when staining of the nucleus and the cytoplasm appeared in red and brown, respectively. All abnormal results were eventually verified by histological examination of the tissue taken from cervical lesions by diagnostic-therapeutic procedure following colposcopic evaluation of cervical lesion topography In the group of cytological smears with ASCUS, the diagnosis was positive in 5 cases (26.3%), negative in 14 (73.7%). In the group with the diagnosis of LSIL, the cytology results were positive in 32 cases (54.2%), negative in 27 (45.8%). In the cytological diagnosis of ASC-H there were 20 positive (87%) and 3 negative (13%) results. Test CINtecPLUS could be a helpful tool in the final diagnosis of cervical abnormality in patients with the cytological diagnosis of ASCUS, LSIL and ASC-H. The combination of conventional cytological test and CINtecPLUS can help create a new procedure algorithm for cases with abnormal or ambiguous cytological screening results. It could be especially useful in a group of young women of childbearing age, when it is common to avoid a more radical treatment of cervical lesions.

Educational needs and causes of false diagnosis of atypical squamous cells of unknown significance at a university hospital.

The entity of atypical squamous cells of undetermined significance (ASCUS) in The Bethesda System 2001 for reporting cervical cytology is characterized by equivocal diagnosis, poor reproducibility and debatable management. This retrospective study was done to analyse the causes of false ASCUS if any and identify the educational needs as part of quality assurance programme. Cervical smears of all ASCUS cases reported over the one-year period were reviewed by the Cytopathologist. Relevant clinical data was retrieved. ASCUS was the most common type of abnormality representing 43.0 % cases among 294 abnormal smears reported during study period. 16.0% cases were found to be non ASCUS on review. The main four causes of over use of ASCUS diagnosis were poor quality smears and cellular atypia associated with Candida infection, atrophy and squamous metaplasia. Educational measures are being undertaken to avoid over diagnosis and improve the patient management.

Triage of cervical cytological diagnoses of atypical squamous cells by DNA methylation of paired boxed gene 1 (PAX1)

Detection of cervical high-grade squamous intraepithelial lesions (HSIL) in patients with equivocal cytological abnormalities, such as atypical squamous cells (ASC) of undetermined significance (ASCUS) or inability to exclude high-grade squamous intraepithelial lesions (ASC-H) is still a challenge. This study tested the efficacy of PAX1 methylation analysis in the triage of cervical ASCUS and ASC-H and compared its performance with Hybrid Capture 2 (HC2) HPV test. A hospital-based case-control study was conducted. Cervical scrapings from patients with ASCUS or ASC-H were used for the quantitative methylation analysis of PAX1 methylation by MethyLight and HPV testing by HC2. Patients with ASC-H or ASCUS with repeated abnormal smears underwent colposcopic biopsy and subsequent therapies. Diagnoses were made by histopathology at a follow-up of 2 years. The efficacies of detecting high-grade lesions were compared. Fifty-eight cervical scrapings with cytological diagnosis of ASCUS (n = 41) and ASC-H (n = 17) were analyzed. One of the 41 (2.4%) ASCUS patients and seven of 17 (41.2%) ASC-H patients were confirmed to have HSIL. After dichotomy of the PMR, PAX1 methylation rates were significantly higher in ASC developing HSIL compared with those developing reactive atypia (87.5% vs. 12.5%, P < 0.001). Testing PAX1 methylation in cervical swabs of patients with ASC confers better sensitivity (87.5% vs. 62.5%) and specificity (98.0% vs. 86.0%) than HC2 HPV testing. We show for the first time that PAX1 hypermethylation analysis may be a better choice than HC2 in the triage of ASCUS and ASC-H.

Infectious Organisms on Papanicolaou Smears Should Not Influence the Diagnosis of Atypical Squamous Cells of Undetermined Significance

Background: Atypical squamous cells of undetermined significance (ASCUS) remain the center of diagnostic controversy and patients’ stress despite recent advances in cervical cancer screening and the introduction of human papilloma virus (HPV) testing. The role of infectious agents in the induction of such changes is not well understood. Aim: We aim at reviewing the effect of the different infectious organisms in Papanicolaou (PAP) smears on the ASCUS diagnosis. Material: 133 ASCUS cases associated with variable infectious organisms (ASCUS-infection group) with secondary HPV testing and appropriate follow-up studies were reviewed. A control group of 310 ASCUS cases without any organisms (ASCUS-only group) was selected for comparison. Results: The ASCUS-infection group had a significantly higher proportion of HPV-positive tests than the ASCUS-only group (p = 0.0027). There was no significant difference on follow-up PAPs and biopsies between the two groups (p = 0.4272). They showed an overall mean of 75% negative, 20% low-grade lesions, and 5% high-grade lesions/carcinoma in situ on follow-up. Conclusion: Our study demonstrates no significant effect of infections on the cytological changes diagnostic of ASCUS. The cytologist should make this diagnosis neglecting any background infections even when predominant.