Brenda Sweeney

Comparison of the effectiveness of two liquid-based Papanicolaou systems in the handling of adverse limiting factors, such as excessive blood.

Excessive blood may compromise gynecologic Papanicolaou (Pap) smears. Liquid‐based cytologic techniques have been developed in part to address this problem. In the current study, conditions of excessive blood were simulated to compare the ability of two liquid‐based systems, ThinPrep® and SurePath™, to satisfactorily process specimens in the presence of this potentially limiting factor.

Performance of the Roche cobas 4800 high-risk human papillomavirus test in cytologic preparations of squamous cell carcinoma of the head and neck.

Determining high‐risk human papillomavirus (HR‐HPV) status of head and neck squamous cell carcinoma (HNSCC) defines a tumor subset with important clinical implications. Cytologic sampling often provides the sentinel or sole diagnostic specimen. The authors assessed the performance characteristics for the Roche cobas 4800 HPV real‐time polymerase chain reaction (PCR)‐based system (cobas) on cytologic specimens of HNSCC compared with standard methods of in situ hybridization (ISH) for HR‐HPV and immunohistochemistry (IHC) for p16 on formalin‐fixed, paraffin‐embedded (FFPE) tissue.

Comparison of BD Surepath and ThinPrep Pap systems in the processing of mucus‐rich specimens

Excessive mucus, as well as blood and inflammation, can be problematic in the processing and screening of liquid‐based cervical Pap preparations by interfering in the process of cell retrieval onto specimen filters or slides. This study compares the capacity of the BD SurePath and ThinPrep liquid‐based Papanicolaou (Pap) tests to handle mucus‐laden specimens.

A superior method for cell block preparation for fine-needle aspiration biopsies.

Cell block (CB) techniques for fine‐needle aspiration biopsies (FNABs) vary. A direct comparison of CB techniques with statistical validation was performed to identify the best method.

Next-Generation Sequencing and Fluorescence in Situ Hybridization Have Comparable Performance Characteristics in the Analysis of Pancreaticobiliary Brushings for Malignancy

Cytological evaluation of pancreatic or biliary duct brushings is a specific, but insensitive, test for malignancy. We compared adjunctive molecular testing with next-generation sequencing (NGS) relative to fluorescence in situ hybridization (FISH) for detection of high-risk neoplasia or malignancy. Bile duct brushings from 81 specimens were subjected to cytological analysis, FISH using the UroVysion probe set, and targeted NGS. Specimens were placed into negative/atypical (negative) or suspicious/positive (positive) categories depending on cytology and negative or positive categories on the basis of FISH and NGS results. Performance characteristics for each diagnostic modality were calculated on the basis of clinicopathologic follow-up and compared in a receiver operating characteristic analysis. There were 33 high-risk neoplasia/malignant strictures (41%) and 48 benign (59%). NGS revealed driver mutations in 24 cases (30%), including KRAS (21 of 24 cases), TP53 (14 of 24 cases), SMAD4 (6 of 24 cases), and CDKN2A (4 of 24 cases). Cytology had a sensitivity of 67% (95% CI, 48%-82%) and a specificity of 98% (95% CI, 89%-100%). When added to cytology, NGS increased the sensitivity to 85% (95% CI, 68%-95%), leading to a significant increase in the area under the curve in a receiver operating characteristic analysis (P = 0.03). FISH increased the sensitivity to 76% (95% CI, 58%-89%), without significantly increasing the area under the curve. These results suggest that ancillary NGS testing offers advantages over FISH, although studies with larger cohorts are needed to verify these findings.

Automated Extraction of Formalin-Fixed, Paraffin-Embedded Tissue for High-Risk Human Papillomavirus Testing of Head and Neck Squamous Cell Carcinomas Using the Roche Cobas 4800 System

CONTEXT -Testing for high-risk human papillomavirus (HR-HPV) in head and neck squamous cell carcinomas (HNSCCs) is important for both prognostication and clinical management. Several testing platforms are available for HR-HPV; however, effective alternative automated approaches are needed. OBJECTIVE -To assess the performance of the automated Roche cobas 4800 HPV real-time polymerase chain reaction-based system on formalin-fixed, paraffin-embedded HNSCC specimens and compare results with standard methods of in situ hybridization (ISH) and p16 immunohistochemistry. DESIGN -Formalin-fixed, paraffin-embedded samples of HNSCC were collected from archival specimens in the Department of Pathology, Massachusetts General Hospital (Boston), and prepared using the automated system by deparaffinization and dehydration followed by tissue lysis. Samples were integrated into routine cervical cytology testing runs by cobas. Corresponding formalin-fixed, paraffin-embedded samples were evaluated for HR-HPV by ISH and p16 by immunohistochemistry. Discrepant cases were adjudicated by polymerase chain reaction. RESULTS -Sixty-two HNSCC samples were analyzed using the automated cobas system, ISH, and immunohistochemistry. Fifty-two percent (n = 32 of 62) of formalin-fixed, paraffin-embedded tumors were positive for HR-HPV by cobas. Eighty-eight percent (n = 28 of 32) of cases were the HPV 16 subtype and 12% (n = 4 of 32) were other HR-HPV subtypes. Corresponding testing with ISH was concordant in 92% (n = 57 of 62) of cases. Compared with the adjudication polymerase chain reaction standard, there were 3 false-positive cases by cobas. CONCLUSIONS -Concordance in HNSCC HR-HPV status between cobas and ISH was more than 90%. The cobas demonstrated a sensitivity of 100% and a specificity of 91% for detection of HR-HPV. Advantages favoring cobas include its automation, cost efficiency, objective results, and ease of performance.

Effects on Cervical Cytology Screening Productivity Associated with Implementation of the BD FocalPoint™ Guided Screener Imaging System

Objectives: Automated screening will become important due to an aging workforce, declining numbers of new cytotechnologists, and the need for increased screening sensitivity in the vaccine era, where high-grade abnormalities will decline. This study documents workload in gynecologic cytology throughput before and after the implementation of the BD FocalPoint™ Guided Screener (GS) System. Study Design: We collected daily screening data from 3 time periods: the 12 months prior to GS implementation, the 6 months immediately after implementation, and the ensuing 7–18 months after implementation. Data was tabulated at the individual and total laboratory levels. Results: In the 6-month period immediately following implementation, productivity increased in 3 of 5 cytotechnologists, as compared to the figures 12 months before implementation. The laboratory increased productivity slightly (+2.4%), with individual changes ranging from –6.9 to +14.7%. In the 7- to 18-month ‘mature’ period after implementation, productivity increased in all 5 cytotechnologists with an average of +15.4%. Individual increases ranged from +6.1 to +26.9%. Conclusions: Overall productivity increased in the period beyond 6 months, and this increase was eventually noted in all personnel. Increased productivity was associated with a short period of learning in which the magnitude of the effect was less than in the mature period.

Advanced practitioner in anatomic pathology: The time has come: Advanced Practitioner in Anatomic Pathology

In this issue of Cancer Cytopathology, Kopp et al from the Mayo Clinic show how properly trained cytotechnologists can go outside their traditional scope of practice to aid the pathologist in the making of tissue microarrays (TMAs) for research. As background, at the Mayo Clinic, demands on pathologists’ time have been increasing, and this is associated with workforce shortages, increasing specimen complexity and interpretation requirements, and decreased reimbursement. As such, important research has been substantially postponed because up to the present, pathologists have been completely responsible for the making of TMAs. TMAs are composite specimens in which small sections (donor cores) from many different paraffin blocks and/or cases are placed into a single block that can then be used to test reagents against multiple samples simultaneously. TMAs are, therefore, very useful in determining the operating characteristics of immunohistochemical and molecular tests. However, to make an optimal TMA, the proper tissue and area within that tissue need to be selected from each case. This selection process means not only that the tissue containing the best, or most representative, target area (eg, the neoplasm or infectious organism) needs to be appropriately identified but also that the selected areas must be free of technical artifacts and other confounding features. This process requires significant morphologic knowledge and skill and diligent screening of the samples under consideration. In this study, cytotechnologists, under the supervision of pathologists, reviewed the study design to understand the research goals for each TMA construction, reviewed the glass slides of the target tissues, and selected the most appropriate blocks for inclusion in the TMA. They then reviewed blocks and slides together to identify the best blocks and areas within the blocks from which the donor cores would be taken. Cytotechnologists then scored the TMA cores for immunohistochemical results. Scoring included the reactivity in the invasive or in situ tumor, stroma, and benign tissues; the type of staining (nuclear, cytoplasmic, or membranous); the quantity of staining (the percentage of cells) and/or the staining intensity; and so forth as needed for each individual TMA research objective. The results of the study showed that appropriately trained cytotechnologists completed all of these tasks successfully. In either complete or percentage spot checks by supervising pathologists, disagreements on tissue selection or scoring were well within the ranges expected from interpathologist correlations and were associated with expected areas of subjective disagreement, such as the best area for obtaining donor cores or stain intensity. Interestingly, cytotechnologists actually performed better at some aspects of the process, most notably in technical assessments of the paraffin block material. The authors concluded their work by noting that research requiring TMAs that had been on hold for years because of a lack of pathologist time was now being completed successfully. This study shows that cytotechnologists have the skill set to successfully assist the pathologist in the performance of research requiring an understanding of pathologic morphology, the screening of large volumes of

Young investigator challenge: Comparison of 2 different methods of manual slide screening in semiautomated gynecologic cytology.

Negative gynecologic cytology cases (ie, those diagnosed as negative for intraepithelial lesion or malignancy) are manually reviewed by 2 methods using semiautomated screening: 1) immediate full slide review (FSR) after fields‐of‐view analysis (FOV) (FOV + FSR), and 2) quality‐assurance/high‐risk, quintile‐directed full manual review (FMR). Data supporting current guidelines were limited. The authors investigated FMR, FOV + FSR, and the review process in general.