Human Papillomavirus Genotyping among Different Cervical Smears in Duhok/Iraq.

Background/Objective: Cervical cancer ranks the second among the most common gynecologic cancers. This study was established to determine the distribution of cervical HPV genotypes among different Pap readings in Duhok/Iraq. Methods: Between January and September-2016, HPV-DNA was tested in 64 women. Genotyping was carried out by the hybridization reverse blot technique. Cervical smears were taken, performed by ThinPrep technique and stained by Pap stain. Results: Twenty six (40.6%) cases were positive for HPV, 12 (46.2%) in normal and 14 (53.8%) in abnormal Pap smears. 39 (19 high-risk and 16 low-risk) genotypes were identified. The high risk group comprised 6 HPV16, 4 HPV18, 2 HPV66, 2 HPV52, 2 HPV39, 1 HPV56, 1 HPV31 and 1 HPV45. The 16 low risk strains encompassed 4 HPV6 strains, 4 HPV71, 2 HPV54 and 2 HPV83, HPV11, HPV61 HPV84, and HPV62. Mixed infections were described in 4 women (6.25%), limited to the NILM, ASC-US and LSIL smears. They included variable admixtures of 7 high risk genotypes, HPV39 (both copies), HPV66 (both copies), HPV52, HPV31, HPV45 and 6 low risk strains: HPV83, HPV6, HPV11, HPV54, HPV62 and HPV71. Conclusions: The higher frequency of HR-HPV than the LR-HPV with identification of 4 mixed cases indicates that our women are at risk of developing cervical cancer. Detection of HR-HPV in NILM and ASC-US smears with restriction of some strains to these 2 categories highlights the great value of HPV genotyping as a surrogate test to pick up unscreened women at risk of developing cervical malignancy particularly when a proper screening program is absent.

Role Of Papanicolaou Smear In Mapping Out Cervical Epithelial Morphology In Women With Infertility: Modified Bethesda Classification.

BACKGROUND: Papinocolaou smear (PAP) smear has a specificity of 98% in detecting early changes in cervical epithelium in women above 21 years, thus its avocation as a screening tool for cervical cancers worldwide. In Pakistan with a lack of awareness in our population as well as socioeconomic conditions there is no such screening program. Infertility on the other hand affects 22% of women in Pakistan and thus one of the most common cause of physician visits. Both cervical cancers and infertility have sexually transmitted diseases (STD's) as common causes with studies reporting ~75% of women have an STD at-least once during their life, presenting with cervical epithelial cell lesions. Thus, the present study was carried out to identify patterns of cervical cell morphology in women with infertility using PAP smear. METHODS: Cervical smears were taken from infertile women and fertile women (n=150), stained with H&E and PAP stains, and graded according to Bethesda Classification 2001. Analysis of data was done by using SPSS version 20 and MS Excel. RESULTS: The mean age of participants was ~29 years. Epithelial cell morphology of these smears showed significant difference among three groups (p value 0.037) with severity in infertile women. Further in the subgroup of secondary infertile women, there were more abnormal smears as compared to primary as well as a higher grade of severity by Pap smear. CONCLUSIONS: Thus, it is concluded that women presenting with some level of infertility are at a higher risk of having cervical epithelial abnormalities.

Association between grade of referral smear and high-grade disease among women with biopsy samples showing cervical intraepithelial neoplasia grade 2.

OBJECTIVE: To determine whether the grade of referral smear reflects the frequency of cervical intraepithelial neoplasia (CIN) grade 3 (CIN3) or worse lesions among patients with CIN2 on punch biopsy. METHODS: In a retrospective study, data were reviewed from women with a punch biopsy sample showing CIN2 and a known referral smear who underwent large loop excision of the transformation zone (LLETZ) between January 1, 2013, and January 1, 2016, at Galway University Hospital, Ireland. Data were analyzed by patient age (≤30 and >30 years), referral smear (low and high grade), and LLETZ histology (≤CIN2 and ≥CIN3). RESULTS: Overall, 264 women were included. LLETZ histology of CIN3 or worse was more common among women with high-grade referral smears (63/144 [43.8%]) than among those with low-grade smears (26/120 [21.7%]; relative risk 2.02, 95% confidence interval 1.37-2.96; P<0.001). Among patients younger than 30 years, underlying CIN3 and above was again more frequent among women with high-grade (44/95 [46.3%]) versus low-grade smears (12/56 [21.4%]; relative risk 2.16, 95% confidence interval 1.25-3.73; P=0.004). No difference was recorded in the older age group. CONCLUSION: Although LLETZ can be performed for a CIN2 biopsy and high-grade smear, consideration should be given among young women (<30 years) with low-grade smears whose biopsy histology is incidentally CIN2.

Debris removal in Pap-smear images.

Since its introduction in the 1940s the Pap-smear test has helped reduce the incidence of cervical cancer dramatically in countries where regular screening is standard. The automation of this procedure is an open problem that has been ongoing for over fifty years without reaching satisfactory results. Existing systems are discouragingly expensive and yet they are only able to make a correct distinction between normal and abnormal samples in a fraction of cases. Therefore, they are limited to acting as support for the cytotechnicians as they perform their manual screening. The main reason for the current limitations is that the automated systems struggle to overcome the complexity of the cell structures. Samples are covered in artefacts such as blood cells, overlapping and folded cells, and bacteria, that hamper the segmentation processes and generate large number of suspicious objects. The classifiers designed to differentiate between normal cells and pre-cancerous cells produce unpredictable results when classifying artefacts. In this paper, we propose a sequential classification scheme focused on removing unwanted objects, debris, from an initial segmentation result, intended to be run before the actual normal/abnormal classifier. The method has been evaluated using three separate datasets obtained from cervical samples prepared using both the standard Pap-smear approach as well as the more recent liquid based cytology sample preparation technique. We show success in removing more than 99% of the debris without loosing more than around one percent of the epithelial cells detected by the segmentation process.

First trial of cervical cytology in healthy women of urban Laos using by self-sampling instrument.

Cervical cancer is the most common cancer in Laos women and a screening programme, even with the PAP smear test (PAP test), has yet to be established for routine use. The Pap test is accepted as the most appropriate for cervical cancer screening in some settings but it is not commonly available in Laos hospitals, because there are few cytopathologists and gynecologists have little experience. As a pilot program, seminars for the PAP test were given in 2007 and 2008, and then PAP tests were carried out using self-sampling instrument (Kato's device) with 200 healthy volunteers in Setthathirath hospital, Laos, in 2008. The actual examination number was 196, divided into class I 104 (53.1%), class II 85 (43.3%), class IIIa 4 (2.0%), class IIIb 1 (0.5%), and class V 1 (0.5%) by modified Papanicolau classification. Four cases had menstruation. There were 6 cases with epithelial cell abnormalities including malignancy. There were 7 cases with fungus and 2 cases with trichomonas in Class II. More than 70% volunteers felt comfortable with the Kato's device and wanted to use it next time, because of the avoidance of the embarrassment and a low cost as compared with pelvic examination by gynecologists. This first trial for PAP test for healthy Laos women related to a hospital found three percent to have abnormal cervical epithelial cells. Therefore, this appraoch using a self-sampling device suggests that it should be planned for cervical cancer prevention in Laos.

Automatic failure in gynecologic cytology proficiency testing. Results from the College of American Pathologists proficiency testing program.

CONTEXT: Automatic failure in gynecologic cytology proficiency testing occurs when a high-grade lesion or carcinoma (HSIL+, Category D) is misinterpreted as negative for intraepithelial lesion or malignancy (Category B). OBJECTIVES: To document the automatic failure rate in 2006 and 2007 from the College of American Pathologists proficiency testing program (PAP PT) and compare them to projected values from 2004. DESIGN: Identify automatic failures from PAP PT in 2006 and 2007 and compare the rates of failure regarding participant and preparation type to validated slides in the College of American Pathologists Interlaboratory Comparison Program in 2004. RESULTS: There were 65 264 participant responses for HSIL+ slides included in this analysis from 2006 and 2007. Overall, 1% (666 of 65 264) of the HSIL+ responses were classified as negative, resulting in automatic failure for the participant. There were significantly fewer automatic failures in 2007 as compared with either 2006 or projected from 2004 data (P < .001). Conventional preparations had a lower automatic failure rate than liquid-based preparations but only for 2006. Both pathologists and cytotechnologists interpreting liquid-based preparations faired better than projected from 2004 data. CONCLUSIONS: The automatic failure rate in PAP PT is lower than expected based on 2004 data from the College of American Pathologists Interlaboratory Comparison Program. Automatic failures are a relatively small component (1% or less) of proficiency testing failures. The rate of automatic failure decreased from 2006 to 2007 and may be due to loss of poor performers in the testing pool, the test-taking environment, or removal of less robust slides from the program.

Pap smear diagnosis using a hybrid intelligent scheme focusing on genetic algorithm based feature selection and nearest neighbor classification.

The term pap-smear refers to samples of human cells stained by the so-called Papanicolaou method. The purpose of the Papanicolaou method is to diagnose pre-cancerous cell changes before they progress to invasive carcinoma. In this paper a metaheuristic algorithm is proposed in order to classify the cells. Two databases are used, constructed in different times by expert MDs, consisting of 917 and 500 images of pap smear cells, respectively. Each cell is described by 20 numerical features, and the cells fall into 7 classes but a minimal requirement is to separate normal from abnormal cells, which is a 2 class problem. For finding the best possible performing feature subset selection problem, an effective genetic algorithm scheme is proposed. This algorithmic scheme is combined with a number of nearest neighbor based classifiers. Results show that classification accuracy generally outperforms other previously applied intelligent approaches.

Appeals in gynecologic cytology proficiency testing: review and analysis of data from the 2006 College of American Pathologists gynecologic cytology proficiency testing program.

CONTEXT: In 2006, 9643 participants took the initial College of American Pathologists (CAP) Proficiency Test (PT). Failing participants may appeal results on specific test slides. Appeals are granted if 3 referee pathologists do not unanimously agree on the initial reference diagnosis in a masked review process. OBJECTIVES: To investigate causes of PT failures, subsequent appeals, and appeal successes in 2006. DESIGN: Appeals were examined, including patient demographic information, Centers for Medicare and Medicaid Services category (A, B, C, or D), exact reference diagnosis, examinees per appeal, examinee's Centers for Medicare and Medicaid Services category, referee's Centers for Medicare and Medicaid Services category, slide preparation type, and slide field validation rate. RESULTS: There was a 94% passing rate for 2006. One hundred fifty-five examinees (1.6%) appealed 86 slides of all preparation types. Forty-five appeals (29%) were granted on 21 slides; 110 appeals (72%) were denied on 65 slides. Reference category D and B slides were most often appealed. The highest percentage of granted appeals occurred in category D (35% slides; 42% of participants) and the lowest occurred in category B (9% slides; 8% of participants). The field validation rate of all appealed slides was greater than 90%. CONCLUSIONS: Despite rigorous field validation of slides, 6% of participants failed. Thirty percent of failing participants appealed; most appeals involved misinterpretation of category D as category B. Referees were never unanimous in their agreement with the participant. The participants and referees struggled with the reliability and reproducibility of finding rare cells, "overdiagnosis" of benign changes, and assigning the morphologically dynamic biologic changes of squamous intraepithelial lesions to static categories.

The accuracy of the Papanicolaou smear in the screening and diagnostic settings.

OBJECTIVE: We evaluated the performance of the Papanicolaou smear in screening and diagnostic settings. STUDY DESIGN: We analyzed Papanicolaou smear results of 1,850 women recruited into a clinical trial to evaluate an emerging technology for the detection of cervical cancer. Screening and diagnosis groups were based on the history of previous Papanicolaou smear results. We calculated sensitivities, specificities, positive and negative likelihood ratios (LR+ and LR-), receiver operating characteristic curves, and areas under the receiver operating characteristic curve (AUC). RESULTS: In the screening group, by defining disease as cervical intraepithelial neoplasia (CIN) 2,3/cancer or worse and using high-grade squamous intraepithelial lesion (HSIL) as the test cutpoint, the AUC was 0.689, and the LR+ and LR- were 39.25 and 0.67, respectively. In the diagnosis group, the AUC was 0.764, and the LR+ and LR- were 3.79 and 0.56, respectively. By defining disease as human papillomavirus/CIN 1 or worse and HSIL as the test cutpoint, the AUC was 0.586, and the LR+ and LR- were 17.01 and 0.92 in the screening group; in the diagnosis group, the AUC was 0.686, and the LR+ and LR- were 2.77 and 0.75, respectively. CONCLUSIONS: In a screening setting, a Papanicolaou smear result of HSIL or worse is 39 times more likely in a patient with CIN 2,3/cancer than in a patient without it. This compares to 4 times more likely in the diagnostic setting. The magnitude of the positive likelihood ratio observed in the screening group indicated that abnormal Papanicolaou smear results obtained in the screening setting should have more impact on clinical decision making than those from results obtained in the diagnostic setting.

Is this the end of the line for the moderate dyskaryotic smear?

OBJECTIVE: The British Society for Clinical Cytology has recently proposed that the terminology for cervical smear reporting is to be changed from a 3-tier system (mild, moderate, severe dyskaryosis) to a 2-tier system of low-grade and high-grade dyskaryosis. This modification eliminates the central category of moderate dyskaryosis which would be incorporated into the high-grade group. The aim of this study was to investigate the role of the moderate dyskaryotic smear in clinical practice. MATERIALS AND METHODS: A retrospective review of all women who were referred for colposcopy because of a moderate dyskaryotic smear was carried out for a 6-month period. Data collected included colposcopic impression, procedure performed and final histopathology. Two cytologists who were unaware of the original smear report were asked to reclassify these smears using the new 2-tier system. Their findings were compared with the documented colposcopic and histopathology results. RESULTS: One hundred women with moderate dyskaryotic smears were referred for colposcopy during the study period. Most of these were reclassified as high-grade dyskaryosis using the new system. Fifty-six (72%) of the moderate dyskaryotic smears which were correctly regraded as high grade by cytologist 1 were found to have cervical intraepithelial neoplasia 2/3 on final histopathology, whereas for cytologist 2, 66 (68%) were found to have high-grade cervical intraepithelial neoplasia. CONCLUSION: There is no clinical benefit in retaining the term moderate dyskaryosis. This study emphasizes the need for a uniform 2-tier system.

Clinical relevance of atypical squamous cells of undetermined significance by the 2001 bethesda system: experience from a cervical cancer high incidence region.

The aim of this study was to evaluate the underlying lesions and factors predicting cervical intraepithelial neoplasia (CIN) 2+ in women who had 'atypical squamous cells of undetermined significance' (ASC-US) on cervical cytology in the region with a high incidence of cervical cancer. This study was prospectively conducted at Chiang Mai University Hospital, Chiang Mai, Thailand. All women with ASC-US cytology undergoing colposcopic evaluation between October 2004 and August 2008 were recruited. During the study period, 208 women were enrolled. Mean age was 44.4 years. The histopathologic results at the initial evaluation were as follows: CIN 2-3, 21 (10.1%); adenocarcinoma in situ, 3 (1.4%); cancer, 5 (2.4%); CIN 1, 26 (12.5%); and no lesions, 153 (73.6%). Multivariate analysis revealed that nulliparity (adjusted odds ratio [aOR] =4.09; 95% confidence interval [CI] = 1.04-16.10) and current oral contraceptive use (aOR=2.85; 95%CI= 1.14-7.15) were independent predictors for having CIN 2+ at the initial colposcopy. At the median follow-up time of 6.7 months, CIN 2-3 lesions were additionally detected in 2 women. In conclusion, ASC-US cytology in our population has a relatively high prevalence of underlying invasive carcinoma. Nulliparity and current oral contraceptive use are independent predictors for harboring CIN 2+.

An interactive computer program for teaching residents pap smear classification, screening and management guidelines: a pilot study.

OBJECTIVE: To assess the feasibility, utility and resident attitudes towards an interactive, Web-based computer learning program. STUDY DESIGN: Pretest/ posttest evaluation of an interactive curriculum to teach current recommendations for cervical cancer screening and classification and management of abnormal Pap smears. Thirty-five Obstetrics and Gynecology (OB/ GYN) residents were encouraged to complete a Web-based computer program that consisted of a pretest, 9 educational sections (3 didactic and 6 case based), a posttest and an exit survey. Clinical cases involved virtual case management of patients' cytologic, histologic and colposcopic images. Mastery was set at 82% (or 23/28 correct answers). RESULTS: Thirty-one of the 35 residents (89%) completed the curriculum. No resident demonstrated mastery (82%) on the pretest; 15 of 31 residents demonstrated mastery on the posttest (p = 0.04). Mean improvement for all residents was 9 additional questions answered correctly. All the residents thought that the Web site was useful and covered all the learning objectives and would recommend it to other residents. CONCLUSION: A program to teach current cervical cancer screening, classification and abnormal Pap smear management guidelines was useful, feasible and well accepted by residents in this population.

2006 consensus guidelines for the management of women with abnormal cervical screening tests.

OBJECTIVE: To provide revised evidence-based consensus guidelines for managing women with abnormal cervical cancer screening test results. PARTICIPANTS: A group of 146 experts, including representatives from 29 professional organizations, federal agencies, and national and international health organizations, met in Bethesda, MD, September 18-19, 2006, to develop the guidelines. MAJOR CHANGES IN GUIDELINES: The core recommendations for managing women with atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesions were changed minimally. Postcolposcopy management for women with these cytological abnormalities is now identical. Management recommendations for these conditions did change for "special populations," such as adolescents for whom a more conservative approach incorporating cytological follow-up for 2 years was approved. Core recommendations for managing women with high-grade squamous intraepithelial lesions and atypical glandular cells also underwent only minor modifications. More emphasis is placed on immediate "screen-and-treat" approaches when managing women with high-grade squamous intraepithelial lesion. Testing for high-risk human papillomavirus DNA is incorporated into the management of women with atypical glandular cells after their initial evaluation with colposcopy and endometrial sampling. The 2004 Interim Guidance for the use of human papillomavirus DNA testing as an adjunct to cervical cytology for screening in women 30 years and older was formally adopted with only very minor modifications. CONCLUSIONS: The 2006 Consensus Guidelines reflect recent data from large clinical trials and advances in technology and are designed to assist clinicians of all subspecialties.

Evaluation of p16INK4a in cervical lesion of premenopausal and postmenopausal women.

Pap smears of postmenopausal women are often misdiagnosed because of the difficulty in distinguishing atrophic epithelial cells groups only by morphological criteria. In this study we investigated the diagnostic application of immunocytochemical staining of p16INK4a on conventional Pap smear. A total of 137 cervical specimens were enrolled in this study, of which 77 and 60 cervical smears were taken from premenopausal and postmenopausal women, respectively. Two cervical smears were taken simultaneously in 68 women, one for conventional cytology and the other for immunostaining. Additional 69 cervical smears were taken from the archive, decolorized and then used for immunostaining. In premenopausal women 1 out of 14 (7.1%) with negative cytology, 7 out of 24 (29.2%) with low grade squamous intra-epithelial lesion (LSIL), all 35 (100%) with high grade squamous intraepithelial lesion (HSIL) and all 4 (100%) with squamous cell carcinoma (confirmed by histopathology) had positive staining to p16INK4a. In postmenopausal women p16INK4a positivity was observed in 4 out of 7 (57.1%) cases of LSIL, 12 out of 14 (85.7%) cases of HSIL and all 4 out of 5 (80%) different cases of carcinoma (1 cervical adenosquamous carcinoma and 3 cervical squamous cell carcinoma in situ confirmed by histopathology), but none of 34 smears with normal cytology. Twenty smears with normal cytology chosen for the negative control in this study were from the group of postmenopausal women and were as expected negative for p16INK4a immunostaining. In the group of postmenopausal women, 16 out of 60 (26.7%) cases the cytological diagnosis was established on the basis of pl6lNK4a immunostaining as being HSIL. From our preliminary study on a limited number of samples, we can however conclude that pl6INK4a immunostaining is a very useful tool for cytological diagnosis enabling to distinguish HSIL from normal, reactive or inflammatory changes.

Modified classification of Gram-stained vaginal smears to predict spontaneous preterm birth: a prospective cohort study.

OBJECTIVE: The purpose of this study was to identify women at risk of spontaneous preterm birth through a 4-category Gram-stained vaginal smear method, not restricted to bacterial vaginosis. STUDY DESIGN: This was a prospective cohort study correlating Gram-stained vaginal smears in early pregnancy with spontaneous preterm birth. Smears were categorized as 'normal,' 'bacterial vaginosis-like,' 'grade I-like' (atypical gram-positive rods) or 'purulent grade I' (lactobacilli-dominated smears showing heavy leukorrhea of unknown cause). RESULTS: Normal microflora were associated with a 4-fold decreased risk (95%CI 0.1-0.6, P < .001) of spontaneous preterm birth and an abnormal Gram stain with an overall adjusted odds ratio of 5.2 (95%CI 1.8-14.5, P < .001). The sensitivity of vaginal smear diagnosis for preterm birth increased from 25% with conventional scoring up to 70% with these modified criteria. CONCLUSION: Accounting for atypical gram-positive bacteria and neutrophils on Gram-stained vaginal smears may identify a larger proportion of women at risk of preterm birth compared to diagnosis of bacterial vaginosis alone.

Organisation of cervical cytology screening in Croatia: past, present and future.

This presentation highlights strengths and weaknesses of cervical cytology screening in Croatia, with particular reference to the opportunistic screening, the use of conventional Papanicolaou (Pap) test and the analysis of some organizational, educational and performance issues that are associated with it. Its aim is to propose measures to improve the efficacy of cervical cytology screening in order to reduce cervical cancer mortality. Currently, in excess of 450,000 Pap tests/ year are examined at 35 laboratories scattered throughout the country. All of these laboratories use standard operating procedures including internal and external quality control. They employ a total of 68 cytologists and 91 cytotechnologists. The sensitivity of cervical screening in Croatia is 90.0%, specificity 98.6%, positive predictive value 92.3%, negative predictive value 98.1% and overall diagnostic accuracy 97.2%. The high diagnostic accuracy of cervical cytology is attributed to the long-standing tradition of education and training of cytologists (postgraduate MSc course since 1967, independent residency since 1974) and cytotechnologists (since 1968). This tradition spanning more than half a century means that today in Croatia there is a developed network of cytology laboratories staffed by highly competent cytologists and trained cytotechnologists. The high accuracy of cancer detection through Pap tests provides strong evidence in support of cervical cytology screening remaining the basic method of prevention for cervical carcinoma. However, some modifications to the current situation are needed. These relate primarily to opportunistic screening. The current screening coverage rate is 68%, although there is capacity, which would allow for all women at risk, i.e. those aged 25-64, to be screened once in three years. The screening coverage relates mainly to those women visiting gynecological out patient clinics for unrelated conditions. A properly organized and controlled national screening programme should replace this. This should be accompanied by the introduction of alternative, highly sensitive methods of sample collection and preparation, such as are available through the introduction of new technologies, e.g. liquid based cytology.