Immunohistochemical evaluation of CON6D/B5: a new CD30 monoclonal antibody.

Immunohistochemical evaluation of CD30 expression is commonly performed in the assessment of hematopoietic disorders and germ cell tumors. A new monoclonal antibody, CON6D/B5, directed against the CD30 antigen has become commercially available for use in formalin-fixed paraffin-embedded tissue samples. The performance characteristics of CON6D/B5 were tested and compared with the well-characterized monoclonal CD30 antibody Ber-H2. Among the various neoplasms analyzed, there was complete concordance between the results of CON6D/B5 and Ber-H2 staining. Similar to Ber-H2, CON6D/B5 immunoreactivity was observed in all cases of classical Hodgkin lymphoma, anaplastic large cell lymphoma, and embryonal carcinoma, with no staining detectable in any of the other various tumors evaluated, including several neoplasms earlier reported as showing CD30 positivity by other investigators. The labeling intensity was much stronger with CON6D/B5 compared with Ber-H2, although no significant differences were observed with regard to the numbers of positively labeled cells in individual cases. The monoclonal antibody CON6D/B5 offers a suitable alternative to Ber-H2 for the immunohistochemical detection of CD30 expression.

The use of virtual microscopy for proficiency testing in gynecologic cytopathology: a feasibility study using ScanScope.

CONTEXT: ScanScope software can digitize entire cytology slides. OBJECTIVE: To test the feasibility of using virtual microscopy methods and "virtual Papanicolaou tests" for proficiency testing (PT) in gynecologic cytopathology. DESIGN: Two PT exercises were conducted using virtual microscopy. Five cytopathologists and 1 cytotechnologist interpreted images using 2 different schema as follows: (1) the College of American Pathologists graded diagnostic codes (CAP-GDCs) and (2) the Center for Medicare and Medicaid Services test scoring categories (CMS-TSCs). The number of diagnostic errors using the CAP-GDCs and the CMS-TSCs, the mean length of time spent diagnosing each case, and the impressions by the users regarding the facility of the technology and image quality were studied. RESULTS: In the first PT exercise, the participants provided incorrect diagnoses in 4 to 8 of the 10 test cases using the CAP-GDCs and in 1 to 4 of the 10 test cases using the CMS-TSCs. In the second PT exercise, the number of errors decreased to 1 to 6 using the CAP-GDCs and to 0 to 6 using the CMS-TSCs. The results did not achieve statistical significance. The mean time of 9.4 minutes spent per case in the second PT exercise was significantly shorter than the 14.4 minutes spent per case in the first PT exercise (P < .001). The ease of use of the software and the image quality were scored by all participants as 3+ or as 4+. CONCLUSIONS: This preliminary study shows that virtual microscopy and virtual Papanicolaou tests prepared using ScanScope may provide effective tools for PT. Technical issues that require further investigation are discussed.

The Anal Pap Smear: Cytomorphology of squamous intraepithelial lesions.

BACKGROUND: Anal smears are increasingly being used as a screening test for anal squamous intraepithelial lesions (ASILs). This study was undertaken to assess the usefulness and limitations of anal smears in screening for ASILs. METHODS: The cytomorphological features of 200 consecutive anal smears collected in liquid medium from 198 patients were studied and findings were correlated with results of surgical biopsies and/or repeat smears that became available for 71 patients within six months. RESULTS: Adequate cellularity was defined as an average of 6 or more nucleated squamous cells/hpf. A glandular/transitional component was not required for adequacy. Dysplastic cells, atypical parakeratotic cells and bi/multinucleated cells were frequent findings in ASIL while koilocytes were infrequent. Smears from LSIL cases most frequently showed mildly dysplastic and bi/multinucleate squamous cells followed by parakeratotic cells (PK), atypical parakeratotic cells (APK), and koilocytes. HSIL smears contained squamous cells with features of moderate/severe dysplasia and many APKs. Features of LSIL were also found in most HSIL smears. CONCLUSIONS: In this study liquid based anal smears had a high sensitivity (98%) for detection of ASIL but a low specificity (50%) for predicting the severity of the abnormality in subsequent biopsy. Patients with cytologic diagnoses of ASC-US and LSIL had a significant risk (46-56%) of HSIL at biopsy. We suggest that all patients with a diagnosis of ASC-US and above be recommended for high resolution anoscopy with biopsy.

Correlation of manual screening and automated (AutoPap 300) analysis of conventional cervicovaginal smears with HPV typing using Digene HC II assay.

Efforts to improve the accuracy and efficacy of diagnosis of cervical cancer and its precursor lesions have prompted the development of new technologies, including several automated screening systems and human papillomavirus (HPV) hybrid capture assay. In this study of 89 conventional cervicovaginal (Papanicolaou [Pap]) smears, diagnoses based on manual screening are correlated with analyses by the AutoPap 300 primary screening system and results of the Digene HPV hybrid capture II assay. We found that (1) the AutoPap system did not "miss" (place in the "no further review" category) any of the high-risk HPV DNA positive cases, (2) quintile assignments by the AutoPap system did not reliably predict the presence or absence of high-risk HPV DNA, (3) cases of low-grade squamous intraepithelial lesion (LGSIL) were associated with high-risk rather than low-risk HPV genotypes, (4) diagnoses of atypical squamous cells of undetermined significance (ASCUS) and above were more frequent in smears from patients whose cervicovaginal specimens tested positive rather than negative for high-risk HPV DNA (37 of 45 vs 14 of 44) by the Digene HPV hybrid capture II assay, and (5) there was a high incidence of high-risk HPV DNA among women whose smears did not show cytomorphologic changes of high-grade squamous intraepithelial lesion (HGSIL). These findings emphasize that high-risk HPV DNA is an indication of risk for HGSIL, not the equivalent for it.

Virtual microscopy as a tool for proficiency testing in cytopathology: a model using multiple digital images of Papanicolaou tests.

BACKGROUND: Modern digital cameras can acquire images from cytologic slides at sufficient resolution to allow for digital enlargement and scrolling on a video monitor, allowing for the simulation of microscopy using a computer. OBJECTIVE: The purpose of this study was to develop a tool for proficiency testing in cytopathology using multiple digital images of Papanicolaou tests. METHODS: Nine images were photographed from each of 10 Papanicolaou tests at x100 optical magnification, 3400 x 2300-pixel resolution, using a light microscope and a digital camera. All images from each case were tiled in a single canvas with Photoshop 4.0 software. Two cytopathologists and 3 cytotechnologists interpreted these "virtual slides" using a computer and graded diagnostic codes (PAP program, College of American Pathologists). Subjects were retested a year later using the glass slides from the same cases and routine microscopy. Both test results, by diagnostic code, were compared with the McNemar test of symmetry. RESULTS: The 5 test subjects provided 42 and 50 correct diagnostic codes by "virtual microscopy" and light microscopy, respectively. No significant asymmetry in results obtained by virtual microscopy and light microscopy was encountered with the McNemar test of symmetry. All test answers were correctly classified by selection series, using both virtual microscopy and light microscopy, and the responses would have been graded as 100% by current PAP program scoring guidelines. This suggests that virtual microscopy could be used for proficiency testing purposes. CONCLUSIONS: A simple virtual microscopy method designed to challenge participants to locate and diagnose cells of interest was effective for the administration of standardized proficiency tests. Virtual microscopy methods that rely on single-plane images to locate and diagnose cells of interest could provide effective proficiency testing tools prior to the development of more computationally intensive systems that represent an entire Papanicolaou test at multiple focal planes.

Endometrial cells and the AutoPap System for primary screening of cervicovaginal Pap smears.

In 1998, the AutoPap 300 received FDA approval for primary screening of conventional cervical smears. As approved, smears categorized as "no further review" and comprising up to 25% of the smears screened by the AutoPap 300 can be reported as negative for malignant and dysplastic cells without screening by a cytotechnologist. We studied 106 conventional cervical smears in which glandular endometrial cells had been identified by manual screening to assess the ability of the AutoPap System (TriPath Imaging, Inc., Burlington, NC) to (1) designate conventional Papanicolaou smears that contain endometrial cells for "review," and (2) stratify smears that contain endometrial cells as more or less likely to be abnormal. Although the number of cases in our study was small, our findings indicate that (1) the AutoPap System is slightly less sensitive than manual screening by experienced cytotechnologists for the detection of endometrial cells in conventional smears, as the System designated for "review" 94.3% of all smears containing endometrial cells, 92.9% of smears reported as atypical glandular cells of undetermined significance (AGUS) or endometrial adenocarcinoma, and 100% of the four smears with subsequently confirmed endometrial adenocarcinomas, (2) ranking of smears into quintiles by the AutoPap System did not provide additional diagnostically useful information with respect to endometrial pathology, (3) the number of endometrial cells in the smears did not correlate with quintile assignment, and (4) for most patients, routine use of the AutoPap System for primary screening of conventional cervical smears is unlikely to contribute to delay in the diagnosis of clinically significant endometrial lesions.