Anal cytology in an HIV-positive population: a retrospective analysis

OBJECTIVE: To retrospectively evaluate cellularity and correlate the presence of columnar cells with specimen interpretation in conventionally prepared anal cytologic smears from an HIV-positive population. STUDY DESIGN: Two cytopathologists and 1 senior cytotechnologist, blinded to the original diagnosis, screened 114 samples from 110 patients collected between 1997 and 2002. One hundred nine males and 1 female were included, age ranging from 23 to 52 years. Discrepancies were reviewed for consensus. The interpretations, cellularity, and presence or absence of columnar cells were noted. The relationships between diagnosis and presence of columnar cells, visible anal lesions, concurrent HIV viral load and CD4+ T-cell counts were assessed. RESULTS: The cytologic findings were as follows: 7, unsatisfactory (6%); 29, negative (25%); 25, atypical squamous cells of undetermined significance (22%); and 53, dysplasia (47%) (42 anal intraepithelial lesion 1 [37%] and 11 anal intraepithelial lesion 2/3 [10%]). Nearly 50% of the smears, 51, showed the presence of columnar cells (45%); 37 of those specimens had some degree of dysplasia (74%). Of the 63 specimens with no columnar cells, 16 (25%) showed dysplasia. Columnar cells were absent from all unsatisfactory specimens. CONCLUSION: A highly significant association between the presence of columnar cells and anal intraepithelial lesion (p<0.001) and a significant association between the presence of columnar cells and atypical cytology when a visible lesion was absent (p=0.0019) were found. No significant relationship was found between the presence/degree of dysplasia and CD4+ T-cell counts or HIV viral load. Lack of clinical follow-up precluded evaluation of the false negative rates in this data set.     

Bethesda 2001. Impact on the reporting of gynecologic cytology

OBJECTIVE: To examine the impact of implementing Bethesda 2001 in one laboratory. STUDY DESIGN: A computer search identified all cervicovaginal specimens evaluated between July 2001 and June 2002. Bethesda 2001 was implemented on January 1, 2002. The rates of specimen adequacy and the frequency of each diagnostic category 6 months before and 6 months after the implementation of Bethesda 2001 were compared. RESULTS: A total of 21,332 cervicovaginal specimens were evaluated during the study period. During the first 6 months, 10,695 specimens were examined; 40% were liquid-based preparations. During the next 6 months, 10,367 specimens were examined; 60% were liquid-based preparations. Prior to the implementation of Bethesda 2001, the percentages of each category were as follows: 74.99% within normal limits, 7.10% reactive/reparative cellular changes (R/R), 10.29% atypical squamous cells (ASC), 0.24% atypical glandular cells (AGC), 3.45% low grade squamous intraepithelial lesion (LSIL), 3.44% high grade squamous intraepithelial lesion (HSIL) and 0.73% unsatisfactory. In addition, 19.00% were classified as "satisfactory but limited by" (SBLB). Following the implementation of Bethesda 2001, the percentages of each category were as follows: 80.09% negative for intraepithelial lesion and malignancy including 6.94% with the qualifier R/R, 10.32% ASC, 0.27% AGC, 4.54% LSIL, 3.44% HSIL and 0.81% unsatisfactory. In addition, 17.40% were satisfactory with a quality indicator (SAT with QI). The incidence of reporting benign endometrial cells in patients over age 40 was the same for both periods. There was a significant decrease in the percentage of specimens classified as SAT with QI when compared to that of specimens classified as SBLB. A statistically significant increase in the percentage of specimens was noted in the category LSIL (P < or = .001) and satisfactory (.005) after implementing Bethesda 2001. No significant changes were noted in other categories. CONCLUSION: Our laboratory experienced some changes in the laboratory statistics of reporting gynecologic cytology after the implementation of Bethesda 2001. Continuous monitoring of reporting trends is indicated to clearly understand the impact of Bethesda 2001 on laboratory statistics.     

Primary screening with liquid-based cytology in an unscreened population in rural China,with an emphasis on reprocessing unsatisfactory samples

OBJECTIVE: To determine the sensitivity and specificity of liquid-based cytology in a high-risk, unscreened population. STUDY DESIGN: A cross-sectional comparative trial of multiple screening techniques. This report focuses on the performance of liquid-based cytology using biopsy as the standard. RESULTS: In total, 1,997 women were screened. All subjects had a minimum of five cervical biopsies. Of the subjects, 4.3% had CIN 2 on biopsy. The ThinPrep Pap with ASCUS as positive had a sensitivity for CIN 2 of 94% and specificity of 78%; the sensitivity for CIN 3 was 98% and for cancer, 100%. The unsatisfactory rate due to bloody specimens was 7.9% (158/1,997). After reprocessing, the unsatisfactory rate was 0.15% (3/1,997). CONCLUSION: Reprocessing effectively recovers ThinPrep samples otherwise unsatisfactory due to blood and mucus. The ThinPrep Pap test is a highly sensitive screening test when used in a high-risk, unscreened population.     

Effectiveness of liquid-based cytology and papanicolaou tests in a low risk population

OBJECTIVE: To compare sensitivity an effectiveness of ThinPrep vs. Pap Test and determine whether the Hybrid Capture System II (HCII) for detection of human papillomavirus (HPV) could increase the sensitivity and effectiveness of cervical screening in a low risk population. STUDY DESIGN: We present a comparative observer, blind, "split sample" study of ThinPrep and Papanicolaou staining in a population of 2026 consecutive women. RESULTS: Conventional Papanicolaou stain detected lesions in 62 cases (3.06%), including 1 atypical glandular cell of undetermined significance (AGUS), 34 atypical squamous cells of undetermined significance (ASCUS) (1.73% of atypias), 20 (0.99%) low grade squamous intraepithelial lesions (LSIL), and 7 (0.35%) high grade squamous intraepithelial lesions (HSIL). ThinPrep detected lesions in 63 cases (3.11%), including 26 (1.28%) ASCUS, 27 (1.33%) LSIL and 10 (0.49%) HSIL. Despite this overall similarity, only 35 had a coincident cytopathologic result by both methods. The kappa concordance index between ThinPrep and Pap was 0.546. Sensitivity and specificity were 74.6 and 45.0 in ThinPrep and 69.5 and 25.0 in Papanicolaou-stained smears. High risk HPV (HR-HP 17) was positive in 13 of ZOASCUS and in 21 of 22 SIL. CONCLUSION: Technologies now widely available such as Thin Prep and viral detection aim to improve accuracy of screening.     

A new,liquid-based cytology technique

OBJECTIVE: To evaluate a new liquid-based cytology technique, Papspin (Thermo Shandon, Pittsburgh, Pennsylvania, U.S.A.). STUDY DESIGN: Three thousand cervical samples were examined. Each cervix was sampled with a Cervex Brush (Roche, Oss, the Netherlands), used first for a Pap smear and afterwards for a Papspin. One cytospin was prepared from each vial. RESULTS: An identical rate of epithelial cell abnormalities (3.8%) was detected with the two methods. Diagnostic concordance was observed in 86% of the 114 cases. Differences in diagnoses occurred in 168 of 3,000 cases (5.6%) concerning fungal infection (22 cases), epithelial cell abnormalities (24 cases) and minimal differences within the nonneoplastic Bethesda category (122 cases). Endocervical cells were absent from 158 Papspins (5.3%) and 66 Pap smears (2.2%), while they were present in the respective Pap smear or Papspin. Seven Papspins were considered "satisfactory, but limited by ..." (SBLB) as compared to 33 Pap smears given the absence of endocervical cells. CONCLUSION: Discordances concerning epithelial cell abnormalities were observed in 24 of 3,000 cases (0.8%). Fungal infections were more easily diagnosed on Papspin. The absence of endocervical cells in 5.3% of Papspins is due to a bias of methodology. Quality improvement was evident on Papspin for SBLB specimens. HPV testing could be performed with good results.     

P16INK4A as an adjunct test in liquid-based cytology

OBJECTIVE: To assess the utility of P16INK4A as an adjunct test in liquid-based cytology in cases with equivocal morphologic changes of high grade squamous intraepithelial lesion (HSIL). STUDY DESIGN: P16INK4A immunoreactivity was investigated in residual ThinPrep material (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) from 30 cases with equivocal diagnoses of HSIL that had corresponding follow-up biopsies. Two control ThinPrep cases were included: 1 HSIL with biopsy-confirmed cervical intraepithelial neoplasia (CIN) 3 and a negative specimen with a corresponding biopsy of squamous metaplasia. The expression of P16INK4A in ThinPrep specimens and corresponding biopsies was scored as previously described. A ThinPrep case was scored positive if it contained > 10 abnormal cells with nuclear and cytoplasmic immunocytochemical staining. Corresponding biopsies were scored as having negative, sporadic, focal or diffuse staining. RESULTS: The P16INK4A antibody assay was positive in 19 of 30 ThinPrep cases (63.3%). Seventeen of the 19 (89.4%) biopsies corresponding to the positively stained ThinPreps also were positive, with a score of at least focal positivity in the dysplastic regions (2 CIN 1, 4 CIN 2, 11 CIN 3; 2 lesions lost in the tissue recut). The assay was negative in 11 ThinPreps (36.6%) and 10 biopsies (33.3%) with tissue confirmation of chronic cervicitis (5), squamous metaplasia (2), CIN 1 (3) and 1 lesion lost in the tissue recut. Seventeen of 18 (94.4%) ThinPreps confirmed as high grade lesions upon biopsy showed P16INK4A positivity. The control HSIL case with a CIN 3 biopsy was diffusely positive for P16INK4A, and the control negative case with biopsy diagnosis of squamous metaplasia was negative. Nondysplastic squamous and metaplastic epithelium in 7 biopsies and nondysplastic squamous or metaplastic cells in ThinPrep cases were negative. Sporadic staining of bacteria, inflammatory cells and endocervical cells was noted. CONCLUSION: ThinPrep cases in the equivocal cytologic category with the corresponding tissue biopsy assayed for P16INK4A expression showed that there was utility for this type of testing. A larger series comparing corresponding ThinPrep and tissue biopsies will be undertaken. The role of HPV infection in these cases will also be explored.     

Use of a liquid-based, thin-layer Pap test in a community hospital. Impact on cytology performance and productivity

OBJECTIVE: To evaluate the direct-to-vial efficacy of the SurePath Pap test (TriPath Imaging, Burlington, North Carolina, U.S.A.) in a community hospital laboratory and to assess its impact on productivity, as measured by Pap test turnaround times (TATs). STUDY DESIGN: A total of 8,771 SurePath Pap tests were compared to 5,055 conventional Pap smears collected and processed over the same 12-month period. SurePath histologic correlation rates were compared to historical correlation rates for conventional Pap smears. Pap test TATs for 3 months prior to implementing SurePath were compared to TATs for the last 3 months of the study, which included an approximate 70:30 ratio of SurePath to conventional Pap tests. Laboratory staffing was unchanged, and mean monthly accessions were relatively constant. RESULTS: SurePath showed statistically significant improvements in the detection of low grade (LSIL) (196%) and high grade (HSIL) (243%) squamous intraepithelial lesions (SILs) relative to conventional Pap smears. The atypical cells of undetermined significance (ASCUS) rate remained nearly constant with SurePath, while the ASCUS/SIL ratio decreased by 68%. Unsatisfactory rates declined 81%. Histologic correlation rates with SurePath increased over those of conventional Pap smears. Pap test mean TATs improved 73% with SurePath. CONCLUSION: Sure-Path detected significantly more cases of LSIL and HSIL than conventional smears without compromising specificity. The ASCUS/SIL ratio and unsatisfactory rate declined dramatically. Pap test TATs also improved markedly with SurePath.     

Outcomes of cervical liquid-based cytology suggesting a glandular abnormality

Objective: To ascertain the positive predictive value of both ?glandular neoplasia (national standard code 6) and borderline change (national standard code 8) in glandular cells in liquid‐based cervical cytology specimens in Cardiff and Vale NHS Trust and to outline the histological outcomes of these cases. Method: Eighty‐nine liquid‐based (Surepath?) cervical cytology cases were retrospectively identified from a 2‐year period (January 2005 to December 2006) and correlated with histopathological diagnoses. Results: Initial punch biopsy histology revealed 18 cases (21%) of cervical glandular intraepithelial neoplasia (CGIN). A further nine cases (10%) of CGIN were identified following local excision or hysterectomy. Ten cases of invasive malignancy were identified: four endocervical adenocarcinomas (all node negative, TNM stage T1b1), five endometrial adenocarcinomas and one squamous cell carcinoma. There were 10 with high‐grade cervical intraepithelial neoplasia (CIN) alone. Women diagnosed with endometrial malignancy presented later with an average age of 64.6 years compared with 34.9 years for endocervical lesions. Taking high‐grade CIN or worse as a positive outcome, the overall positive predictive value (PPV) of glandular abnormalities on cytology (both code 6 and 8) was 58.1% [95% confidence interval (CI) 47.8, 68.4]. PPV for borderline change in glandular cells alone was 24.1% (95% CI 8.5, 39.6) and for ?glandular neoplasia alone 75.4% (95% CI 64.3, 86.5). Conclusion: With our interpretation of the classification, women with cytological diagnoses of glandular neoplasia of the cervix should initially be investigated by local resection rather than punch biopsy, and those with borderline change in glandular cells with repeat cytology.     

Endometrial carcinoma detected with SurePath liquid-based cervical cytology: comparison with conventional cytology

Introduction: Conventional Pap smears (CPS) have little impact on the detection of endometrial carcinoma. Although liquid‐based cytology (LBC) is replacing CPS in the UK, experience with identification of endometrial cancers with this technique is limited. Aim: To compare the accuracy of the SurePath LBC with that of CPS for detection of endometrial cancers. Methods: Our study group comprised SurePath LBC samples reported as atypical endometrial cells and endometrial adenocarcinoma (classified respectively as borderline, code 8 and ?glandular neoplasia, code 6 for the NHS Cervical Screening Programme statistics) in 2004–2005. CPS reported as atypical endometrial cells or adenocarcinoma in 1993–1998 comprised the control group. Histological follow‐up was obtained. Results: Endometrial abnormalities were reported in 95 (0.073%) of 130 352 LBC samples, comprising 75 (0.058%) atypical endometrial cells and 20 (0.015%) endometrial adenocarcinoma reports. Of 409 495 CPS, 117 (0.029%) were diagnosed as endometrial abnormalities, comprising 59 (0.014%) atypical endometrial cells and 58 (0.014%) endometrial adenocarcinoma reports. Thus, the endometrial adenocarcinoma reporting rate was similar in both groups, but that for atypical endometrial cells was higher with LBC (P < 0.001). The positive predictive value for endometrial cancer of endometrial adenocarcinoma and atypical endometrial cell reports in the LBC group was 73.3 and 18.8%, respectively, compared with 42.3 and 6.7% in the CPS group. The endometrial adenocarcinoma patients in CPS group were older (mean age 62.5 years versus 56.5 years) and most (22/25) were symptomatic, whereas most (13/17) patients in the LBC group were asymptomatic at the time of sampling (P < 0.001). Conclusion: SurePath LBC is at least as accurate a method for detecting endometrial cancer as CPS. SurePath LBC demonstrates enhanced identification of endometrial pathology in asymptomatic women in the cervical screening programme.     

Using a quality control approach to define an 'adequately cellular' liquid-based cervical cytology specimen

Objective:  To define a minimum acceptable total squamous cellularity for (ThinPrep^®) liquid-based cervical cytology (LBC) specimens using quality control techniques.
Methods:  Two hundred LBC preparations were made containing varying numbers (<200) of severely dyskaryotic squamous cells and with varying total cellularities.
Results:  Ninety-eight per cent of the LBC preparations that were missed by one or more of three cytoscreeners had fewer than 16 abnormal objects (single dyskaryotic cells or clumps of cells) and 87 dyskaryotic cells. The minimum ratio of dyskaryotic to total squamous cells that, in a preparation of 5000 squamous cells has a probability of at least 0.98 that 87 or more dyskaryotic cells will be present is 1 : 47. Twenty-three preparations diagnosed as abnormal had ratios of dyskaryotic to total squamous cells of between 1 : 2.5 and 1 : 4596. There is thus no feasible minimum acceptable squamous cellularity that will give an acceptable probability of detection of all specimen vials containing abnormal cells in the observed proportions.
Conclusions:  It is suggested that the minimum acceptable cellularity for LBC specimens is set pragmatically by the screening programme to give a feasible percentage of repeat tests.     

Computerized reporting and follow-up of gynecologic cytology

A system utilizing inexpensive computer hardware to produce high-quality cytology report and to provide an extensive, semi-automated follow-up system is described. Key features of the reports generated are emphasized. The system uses the Bethesda System nomenclature for reporting cytologic findings to a wide range of physicians, from general practitioners to gynecologists to subspecialists in gynecologic oncology.     

Non-computer-assisted liquid-based cytology for diagnosis of oral squamous cell carcinoma

Abstract

The development of oral squamous cell carcinoma (OSCC) occasionally follows the neoplastic progression of other premalignant lesions. Although biopsy is the definitive diagnostic method, liquid-based cytology is an adequate method for screening suspicious lesions. We compared liquid-based cytology to histology for diagnosis of OSCC in patients with oral lesions that raised clinical suspicion of malignancy. Our sample consisted of 48 patients. Cytological samples were obtained by scraping the lesion superficially using Cytobrush^®. We conducted cytological and histopathological evaluation of all preparations. We estimated sensitivity and specificity levels as well as positive and negative predictive values. The degree of inter-observer agreement for both methods was assessed using the kappa index. Twenty-eight (58.3%) of the cases finally were diagnosed with OSCC and 20 (41.7%) were determined to be premalignant lesions. We observed eight false negatives and no false positives; OSCC prevalence was 56.5%. The values for diagnostic indices were: sensitivity, 69% (CI 95%, prevalence 51.87); specificity, 100%; positive predictive value, 100%; negative predictive value, 71% (CI 95% 54.82). A kappa index of 0.622 (CI 95% 0.93, 0.39) was observed.     

Follow-up study of atypical glandular cells in gynecologic cytology using conventional Pap smears and liquid-based preparations: impact of the Bethesda System 2001

OBJECTIVE: This study evaluated the impact of the Bethesda System (TBS) 2001 in reporting of atypical glandular cells (AGC) when using conventional Pap smears (CS) and liquid-based cytology preparations (LBC). STUDY DESIGN: Follow-up information for all atypical glandular cells of undetermined significance (AGUS)/ AGC cases encountered in Queen Mary Hospital from July 2000 to June 2004 was analyzed. The difference in percentages associated with certain end points when using different reporting systems and preparation methods were compared. The age trends and time interval between cytologic diagnosis and detection of positive end points were studied. RESULTS: More than half of these cases turned out to be "negative." The majority with "negative" end points belonged to the "not otherwise specified" (NOS) groups (including atypical endometrial cells) in TBS 2001. The connotation of "favor neoplastic" carried a high positive predictive value for significant lesions. Most of the significant outcomes were discovered within the subsequent 6 months. A decreased reporting of "AGC, NOS" and an increased reporting of "atypical endocervical cells, NOS" were noted when using LBC. CONCLUSION: Subcategorization of AGC in TBS 2001 according to cellular origin and risk of malignancy, which is further enhanced by application of LBC, is useful.