Training undergraduate students for rapid on-site evaluation of fine needle aspiration cytology samples using a simulation-based education activity.

INTRODUCTION: Demand for rapid on-site evaluation (ROSE) of fine needle aspiration (FNA) cytology is rising and the role is increasingly being performed by non-medical cytologists. Undergraduate training for cytologists has traditionally focused on laboratory-based procedural activities and their theoretical underpinning, with minimal attention given to communication and other skills required to operate in an interprofessional setting. We evaluated the effectiveness and student reaction to a simulation-based education (SBE) exercise in ROSE designed to fill this void. METHODS: We designed and evaluated an SBE exercise based on FNA ROSE across two tertiary institutions with 79 students. The exercise accurately reproduced the demands on cytologists operating as part of a multi-disciplinary team in a time- pressured environment. FINDINGS: Pre- and post-simulation questionnaires indicated an improvement in technical knowledge related to the procedure. Students' perception of their competence and confidence in their role also improved significantly post simulation. Students uniformly found the exercise engaging and a valuable addition to their curriculum. DISCUSSION: The simulation successfully provided a pseudo-clinical environment that highlighted the realities of practising technical and diagnostic tasks under time pressure and interacting with other health professionals to provide an optimal patient outcome. The exercise is a useful supplement to on-the-job training for ROSE.

Claudin-4 immunohistochemistry is a useful pan-carcinoma marker for serous effusion specimens.

OBJECTIVE: To evaluate the utility of claudin-4 as a pan-carcinoma marker in cell-blocks of effusion specimens and compare results with Ber-Ep4 staining. METHODS: Effusion cell-blocks (n = 284) were stained for claudin-4 and results compared with Ber-Ep4. Cases included 172 metastatic malignancies (137 adenocarcinomas, 20 small cell lung tumours, eight metastatic melanoma, four squamous cell carcinoma, three urothelial cell carcinoma), 49 benign reactive cases and 63 mesotheliomas. RESULTS: All 49 benign effusions were negative. Only 1/63 (1.6%) mesotheliomas was positive for claudin-4. Claudin-4 staining was positive in 131/137 (95.6%) adenocarcinoma cases. Cases negative for claudin-4 included single cases of metastases from breast, colon, stomach, prostate, kidney and ovary. Claudin-4 outperformed Ber-Ep4. Sensitivity (95.6% vs 85.4%), specificity (99.1% vs 86.6%), negative predictive value (94.9% vs 82.9%) and positive predictive value (99.2% vs 88.6%) were all higher for claudin-4 compared with Ber-Ep4, respectively. Only two cases were claudin-4-/Ber-Ep4+. Significantly (P < .0064) more cases of metastatic adenocarcinoma stained positive for claudin-4 (131/137; 95.6%) than Ber-Ep4 (117/137; 86.2%). Claudin-4 staining was present in 15/20 (75%) of neuroendocrine carcinomas, 3/4 (75%) squamous cell carcinoma and 3/3 (100%) urothelial cell carcinoma. All eight cases of melanoma were negative for both claudin-4 and Ber-Ep4. CONCLUSIONS: Claudin-4 staining is a useful addition to IHC panels for effusions specimens with superior performance to Ber-Ep4.

Investigation of lymphoid lesions of the head and neck using combined fine needle aspiration cytology and flow cytometry: Accuracy and pitfalls.

OBJECTIVE: We reviewed the diagnostic utility of combined fine needle aspiration cytology (FNAC) and flow cytometry (FC) in the diagnosis of lymphoid lesions of the head and neck. METHOD: In total, 1402 patients with combined FNAC-FC reports were correlated with follow-up information. Rapid on-site evaluation (ROSE) of cytological specimens was performed in 52% of cases. RESULTS: In total, 211 lymphoid malignancies were identified, including 198 non-Hodgkin lymphoma (NHL) and 13 Hodgkin lymphoma (HL). Accuracy measures for NHL were: sensitivity 95.5%; specificity 99.9%; PPV 99.5%; NPV 99.2%; accuracy 99.3%. Only seven of 13 cases of HL were detected by FNAC-FC. False negative cases included HL (six cases), diffuse large B-cell lymphoma (four), T-cell lymphoma (two), follicular lymphoma (one), marginal zone cell lymphoma (one) and B-cell NHL, not otherwise specified (one). Two false positive results were identified: one immunoblastic hyperplasia reported as suspicious for HL and one case reported as suggestive of NHL that was found to be reactive hyperplasia. Cases collected with ROSE had a significantly lower rate (P < 0.0001) of insufficient cells for FC analysis (7.0%) than cases where ROSE was not performed (16.4%). Sensitivity (P < 0.0001) and NPV (P = 0.0023) were significantly higher for ROSE-collected specimens. None of the false-negative NHL cases had ROSE performed. CONCLUSIONS: FNAC-FC is a highly sensitive and specific test for NHL. Diagnostic errors mostly involved HL, large cell lymphomas and T-cell lymphomas. ROSE results in a significantly higher adequacy rate for FC and higher sensitivity for NHL.

Gata3 Immunohistochemical Staining is A Useful Marker for Metastatic Breast Carcinoma in Fine Needle Aspiration Specimens.

AIMS: The utility of GATA3 immunohistochemistry (IHC) as an aid to the cytological diagnosis of metastatic breast carcinoma in fine needle aspiration (FNA) specimens was investigated. MATERIALS AND METHODS: Cell block sections from 111 FNA cases of metastatic malignancy were stained for GATA3, including metastases from 43 breast and 44 nonmammary adenocarcinomas, 19 melanomas, 4 urothelial carcinomas, and 1 thyroid medullary carcinoma. Sites sampled included lymph nodes (87), bone (8), liver (5), lung (6), superficial masses (4), and pelvic mass (1). RESULTS: Ninety-one percent (39/43) of metastatic breast carcinoma cases were positive for GATA3. All estrogen receptor (ER)-positive were also GATA3 positive cases. The majority (9/14; 64%) of ER-negative and 37% (3/8) of triple-negative cases were positive for GATA3. All nonmammary adenocarcinoma cases were negative with the exception of one case of metastatic pancreatic adenocarcinoma. Metastatic melanoma cases were all negative but 75% (3/4) urothelial carcinomas expressed GATA3. CONCLUSIONS: GATA3 IHC staining is a useful addition to IHC panels for FNA samples in specific settings such as distinguishing metastatic breast from lung carcinoma or melanoma.

External quality assurance in nongynecologic cytology: The Australasian experience.

BACKGROUND: The Royal College of Pathologists of Australasia Cytopathology Quality Assurance Program has operated an external quality assurance program in nongynecologic cytopathology since 1993. Glass slide preparations of a wide range of nongynecologic cases were circulated to approximately 200 cytopathology laboratories in 16 countries. METHODS: General nongynecologic cytology cases were manufactured from residual specimens after routine diagnosis. Fine-needle aspiration (FNA) cases were made by sampling fresh tissue and making direct specimens. The majority of cases consisted of both air-dried and fixed preparations. Results returned to laboratories included illustrated case discussions highlighting diagnostic features, key differential diagnoses, and useful adjunctive tests. RESULTS: The current study reviewed >22,000 results for 123 nongynecologic cases. Cases found to cause the most diagnostic difficulties included serous effusion cases with metastatic carcinoma in a dispersed pattern, well-differentiated carcinoma, and cellular reactive cases; urine specimens with sparse malignant cells; reactive pneumocytes in a bronchoalveolar lavage; breast FNA cases with papillary lesions; gestational specimens; and fibroadenoma. FNA specimens from the lung and thyroid, particularly papillary thyroid carcinoma, generally were well reported. CONCLUSIONS: The use of multiple preparations of the same specimen has allowed interlaboratory comparison, and the quality assurance program has played an educational role as well as informing the laboratory accreditation process. Cancer Cytopathol 2017;125:349-361. © 2017 American Cancer Society.

Fine-needle aspiration cytology of Merkel cell carcinoma-a review of 69 cases.

This study reviewed the clinical presentation, cytologic findings, and the immunophenotype of 69 Merkel cell carcinoma (MCC) cases sampled by fine-needle aspiration (FNA). Demographic and clinical data, the cytology findings, and results of ancillary testing were reviewed. Median patient age was 78 years (37-104) with a 1:1.8 female to male ratio. The most common FNA sites sampled included lymph nodes in the neck, the axillary region, the inguinal region and the parotid gland. Most patients had a history of MCC (68%) and/or non-MCC malignancy (70%). The common cytologic pattern was a cellular smear with malignant cells arranged in a dispersed pattern with variable numbers of disorganized groups of cells. Cytoplasm was scant or absent and nuclei showed mild to moderate anisokaryosis, stippled chromatin, inconspicuous nucleoli, and nuclear molding. Numerous apoptotic bodies were often present. Cell block samples (28 cases) were usually positive for cytokeratins in a perinuclear dot pattern, including 88% of cases with CK20 positivity. CD56 was the most sensitive (95%) neuroendocrine marker on cell blocks and was also positive with flow cytometry in nine cases tested. MCC is most commonly seen in FNA specimens from the head and neck of elderly patients, often with a history of previous skin lesions. Occasional cases present in younger patients and some may be mistaken for other round blue cell tumors, such as lymphoma. CD 56 may be a useful marker in cell block preparations and in flow cytometric analysis of MCC.

GATA3: a promising marker for metastatic breast carcinoma in serous effusion specimens.

BACKGROUND: The usefulness of GATA3 (GATA-binding protein 3 to DNA sequence [A/T]GATA[A/G]) as a marker for metastatic breast carcinoma in serous effusion specimens was investigated. METHODS: Cell block sections from 74 serous effusion specimens (32 ascitic, 2 pericardial, and 40 pleural fluids) were stained with an anti-GATA3 murine monoclonal antibody. The specimens included 62 confirmed metastatic carcinomas from the breast (30 specimens), female genital tract (13 specimens), gastrointestinal tract (7 specimens), lung adenocarcinoma (9 specimens), pancreas (1 specimen), kidney (1 specimen), and bladder (1 specimen). The breast carcinoma cases included 15 ductal carcinomas and 8 lobular carcinomas; the histology subtype was not available for 7 specimens. Twelve cases containing florid reactive mesothelial cells were also stained. The breast carcinoma cases were also stained for mammaglobin and gross cystic disease fluid protein of 15 kilodaltons (GCDFP-15) to compare their sensitivity with GATA3. RESULTS: Positive nuclear staining for GATA3 was found to be present in 90% of metastatic breast carcinoma specimens (27 of 30 specimens). All nonbreast metastatic carcinomas tested were negative with the exception of the single case of metastatic urothelial carcinoma. No staining was observed in any of the benign reactive cases or in benign mesothelial cells present in the malignant cell block preparations. Two cases demonstrated weak positivity of benign lymphoid cells. Staining results were unambiguous because all positive cases demonstrated intense nuclear staining in > 50% of tumor cells. Mammaglobin (57% staining; 17 of 30 cases) and GCDFP-15 (33% staining; 10 of 30 cases) were found to be less sensitive markers of breast carcinoma. If used in a panel, mammaglobin and GCFP-15 staining would have identified only 1 additional case compared with those stained with GATA3. CONCLUSIONS: GATA3 may be a useful addition to immunostaining panels for serous effusion specimens when metastatic breast carcinoma is a consideration.

Performance measures for Australian laboratories reporting cervical cytology: a decade of data 1998-2008.

AIM: Performance measures for Australian laboratories reporting cervical cytology are a set of quantifiable measures relating to the profile and accuracy of reporting. This study reviews aggregate data collected over the ten years in which participation in the performance measures has been mandatory. METHODS: Laboratories submit annual data on performance measures relating to the profile of reporting, including reporting rates for technically unsatisfactory specimens, high grade or possible high grade abnormalities and abnormal reports. Cytology-histology correlation data and review findings of negative smears reported from women with histological high grade disease are also collected. Suggested acceptable standards are set for each measure. This study reviews the aggregate data submitted by all laboratories for the years 1998-2008 and examines trends in reporting and the performance of laboratories against the suggested standards. RESULTS: The performance of Australian laboratories has shown continued improvement over the study period. There has been a fall in the proportion of laboratories with data outside the acceptable standard range in all performance measures. Laboratories are reporting a greater proportion of specimens as definite or possible high grade abnormality. This is partly attributable to an increase in the proportion of abnormal results classified as high grade or possible high grade abnormality. Despite this, the positive predictive value for high grade and possible high grade abnormalities has continued to rise. CONCLUSION: Performance measures for cervical cytology have provided a valuable addition to external quality assurance procedures in Australia. They have documented continued improvements in the aggregate performance, as well as providing benchmarking data and goals for acceptable performance for individual laboratories.

Thin-layer preparations of dithiothreitol-treated bronchial washing specimens.

OBJECTIVE: To evaluate the combined effect of dithiothreitol (DTT) treatment and ThinPrep (TP) (Cytyc Corp, Boxborough, Massachusetts, U.S.A.) processing on bronchial washing specimens. STUDY DESIGN: A total of 431 bronchial washing specimens were initially treated with 0.05% DTT in a 30% methanol solution. After centrifugation, 1 TP slide and 2-4 conventional cytospin or smear preparations (CPs) were prepared. The reports of both preparations were compared in all cases. All 48 abnormal cases and 52 consecutive negative cases were also compared for cellular composition, distribution of the cells, ease of interpretation and overall preparation quality. Screening time was recorded for 20 of the cases. RESULTS: The diagnostic accuracy of one TP slide appeared comparable to that of 2-4 CPs. The TP slide was assessed to be equal or superior in overall quality to CP in 85% of 100 cases of paired specimens. The cleaner background and smaller cellular area of TP slides significantly reduced the screening time. Mucolysis and specimen homogenization were not always optimal, occasionally resulting in uneven subsampling and poorly cellular TPs. However, in general, TP slides were considered superior to CPs in overall quality. CONCLUSION: Improvement in specimen quality and reduced screening time have to be balanced against the high cost of consumables with the TP technique.