American journal of clinical pathology最新文献

Objective: We examined the utility of preferentially expressed antigen in melanoma (PRAME) immunohistochemistry (IHC) in distinguishing breast angiosarcoma from benign or atypical vascular lesions (AVLs) of the breast.

Methods: There were 26 breast angiosarcomas (24 radiation related and 2 primary), 6 radiation-related AVLs of the breast, and 16 other benign vascular lesions of the breast retrieved from our institutional archive and stained with PRAME IHC.

Results: Twenty-four of 26 (92%) breast angiosarcomas were positive for PRAME with moderate to strong staining intensities (median HScore, 255 [range, 90-300]), while 0 of 22 benign or atypical vascular lesions of the breast were PRAME positive (sensitivity, 92.3% [95% CI, 75.9%-98.6%]; specificity, 100% [95% CI, 85.1%-100%]); c-MYC was positive in all angiosarcoma cases but showed weak staining in several cases (median HScore, 155 [range, 30-280]). PRAME was negative in 1 of 2 primary angiosarcomas and negative in only 1 of 24 secondary angiosarcomas of the breast. None of the benign or atypical vascular lesions was stained with PRAME.

Conclusions: It appears that PRAME expression occurs in a high proportion of radiation-related angiosarcomas and not in benign breast vascular lesions or AVLs. PRAME IHC is diagnostically useful in distinguishing benign from malignant vascular proliferations in patients with a history of radiation to the breast.

Objective: To determine trends in system and scanner downtime in our institution's digital pathology pipeline since its implementation.

Methods: Scanner and system downtime data were tabulated from a period beginning in 2017 and ending in 2022. Downtime events were categorized based on their etiology, such as image management system related for the overall system or hardware vs software related for the scanner.

Results: The maximum scanner downtime consisted of 36 events and occurred in the first quarter of 2019; most of this downtime was attributed to hardware issues. The average scanner downtime per quarter was 350.7 hours. Multifactorial events tended to last longer than single events. System downtime was mostly due to the image management system. Full-system downtime occurred from 2017 through 2019; since then, full-system downtime has essentially been replaced with partial downtime.

Conclusions: Scanner downtime was mostly due to hardware, while system downtime was mostly caused by issues with the image management system. With experience, our institution mitigated the impact of technological difficulties, significantly reducing the number of downtime events since the implementation of digital pathology in 2017.

Objective: The aim of this study was to evaluate a large cohort of gallbladder amyloid cases to determine clinical and morphologic features.

Methods: Cholecystectomy specimens (N = 118) typed using proteomics-based techniques between 2008 and 2023 were identified. Clinical and morphologic features were reviewed.

Results: Six amyloid types were identified: ATTR (n = 63, 53.4%), AL (n = 46, 39.0%), AA (n = 4, 3.4%), AApoA1 (n = 2, 1.7%), ALECT2 (n = 2, 1.7%), and AEFEMP1 (n = 1, 0.8%). Amyloidogenic mutations were detected in 3 ATTR cases and 2 AApoA1 cases. Morphologic review (n = 26) revealed perimuscular vessel involvement in all cases. Amyloidosis was an unexpected diagnosis first made on the cholecystectomy specimen in half of the patients with clinical information (n = 10). All 9 patients with follow-up had evidence of systemic disease. In 2 patients, cholecystic involvement was initially missed and only retrospectively identified after the diagnosis of cardiac amyloidosis.

Conclusions: In patients with clinical data, amyloidosis was often unexpected, the gallbladder was commonly the first tissue sampled with amyloidosis, and all patients had systemic disease. Thorough review of cholecystectomy specimens with careful inspection of perimuscular vessels, coupled with a low threshold for ordering Congo red stain in elderly individuals and amyloid typing using a robust method such as proteomics, can prevent a delay in amyloid diagnosis and management.

Objective: We hypothesized that a set of immunohistochemistry (IHC) stains could be used to distinguish Burkitt lymphoma (BL), the quintessential B-cell lymphoma with a germinal center B-cell (GCB) dark zone (DZ) expression signature, from diffuse large B-cell lymphoma, not otherwise specified (DLBCL, NOS). This might also be applicable to high-grade B-cell lymphomas (HGBCLs) with MYC and BCL2 rearrangements (double-hit lymphomas [DHLs]) and triple-hit lymphomas (THLs).

Methods: A 5-marker IHC algorithm was designed from gene lists that distinguish physiologic DZ from light zone GCBs.

Results: In training and validation cohorts, we distinguished BL from DLBCL, NOS with high sensitivity and specificity. Because DHLs/THLs are enriched for the gene expression DZ signature (DZsig), we evaluated 19 DHLs/THLs and 4 HGBCLs, NOS. Most (83%) cases were IHC DZ. The NanoString DLBCL90 assay was performed on 34 cases to correlate IHC DZ results with the molecular DZsig. The IHC DZ call was significantly associated with the DZsig (P = .0011). The sensitivity and specificity of IHC to recognize DZsig+ cases among DLBCL, NOS and DHLs with BCL2 rearrangements/THLs were 91% and 100%, respectively.

Conclusions: The IHC DZ algorithm can support a diagnosis of BL and identifies MYC-BCL2 DHLs/THLs with a molecular DZsig.

Objective: Intratumoral heterogeneity (ITH) of Ki67 expression reflects the proliferative diversity of breast cancer (BC) cells and has been associated with disease progression. Quantification of Ki67 ITH using Haralick entropy metric from digital image analysis (DIA) has been reported as an independent predictor of breast cancer-specific survival (BCSS); however, its reproducibility across DIA platforms and dependence on tumor tissue sampling have not been investigated.

Methods: Whole-slide images of Ki67-stained tumor sections from 254 patients with ER+/HER2- BC were analyzed independently using HALO and Aiforia DIA platforms. The DIA outputs were subsampled using hexagonal grids to compute Ki67 Haralick entropy. Reproducibility was tested across DIA platforms and under simulated surgical excision and core biopsy scenarios. Lastly, the impact on prognostic modeling for BCSS was assessed.

Results: Haralick entropy demonstrated strong Ki67 ITH cross-platform reproducibility. For prognosis, it provided stronger model performance than conventional Ki67% metrics and independently predicted worse BCSS alongside lymph node involvement. Its prognostic value remained consistent across simulated sampling scenarios.

Conclusions: Ki67 Haralick entropy is a reproducible and robust image-derived ITH metric in ER+/HER2- BC. It demonstrated improved prognostic modeling performance compared to conventional Ki67% across 2 different DIA platforms and sampling conditions, supporting its potential for clinical implementation.

Objective: This study evaluates an automated fluorescence resonant energy transfer (FRET)-based ADAMTS13 activity assay on the Ceveron S100 instrument for the diagnosis of thrombotic thrombocytopenic purpura. It addresses the challenge of high background fluorescence (HBF), a known concern from our manual FRET assay, and proposes strategies to minimize erroneous results.

Methods: We compared FRET-Ceveron results with FRET-Manual (n = 100) and Technozym (Technoclone) enzyme-linked immunosorbent assay (ELISA) (n = 52) using retrospective and prospective patient samples collected throughout 2024, alongside proficiency samples and standards with assigned values (n = 24). We analyzed 7 spiked samples with HBF and 14 patient samples exhibiting HBF while exploring predilution methods. Over 200 FRET-Ceveron reactions were examined to identify abnormal patterns and establish thresholds for HBF interference.

Results: The FRET-Ceveron assay demonstrated a strong correlation (r² > 0.97) with Technozym ELISA, FRET-Manual, and target results. It successfully detected critically low ADAMTS13 levels (<10%) across various sample types (n = 15). While HBF affected both FRET methods, FRET-Ceveron displayed greater tolerance to HBF. No significant difference was found in FRET-Ceveron result accuracy for initial carbon nanotubes (CNTs) up to 1100 (P = .39), but significant differences were observed when CNTs exceeded 1100 (P = .02). Predilution effectively reduced HBF (P < .05), validating the results confirmed by Technozym ELISA.

Conclusions: The fully automated FRET-Ceveron assay is a rapid and accurate method for ADAMTS13 testing, and it is particularly effective when a normal reaction pattern is observed (initial CNTs ≤1000 with a good linearity in reaction tracing during 7- to 22-minute measurements). New sample collection is preferred in the presence of HBF, with predilution as a viable option.

Objective: Chlamydia trachomatis and Neisseria gonorrhoeae present substantial public health challenges. Accurate diagnostic testing is essential to prevent misdiagnosis and unnecessary treatment. Although nucleic acid amplification tests offer excellent performance, they are not infallible. This study sought to evaluate the semiquantitative utility of relative light unit (RLU) values from the Hologic Aptima Combo 2 Assay to improve the diagnostic accuracy of testing for C trachomatis and N gonorrhoeae.

Methods: Data were analyzed from January 2021 to December 2021. Manufacturer guidelines define results as positive if the RLU value is above 100 for C trachomatis only, above 150 for N gonorrhoeae only, and above 250 for dual C trachomatis and N gonorrhoeae detection; equivocal if the RLU value is 25 to 99 for C trachomatis, 60 to 149 for N gonorrhoeae, and 85 to 249 for both; and negative if the RLU value is below 25 for C trachomatis, below 60 for N gonorrhoeae, and below 85 for both. Manufacturer guidance recommends repeat testing only for equivocal results. In contrast, the University of Wisconsin University Hospital adopted a modified criterion, classifying all results with an RLU value at or below 900 as equivocal and requiring repeat testing.

Results: In this retrospective review of 20 875 Aptima Combo 2 assays performed from January to December 2021, 7 patients had initial positive results, with RLU values at or below 900. Of these, 5 were ultimately determined to be false positives.

Conclusions: These findings demonstrate that expanding the definition of equivocal results to include low positive RLU values (≤900) increases identification of false positives with minimal additional repeat testing. This modified approach may improve diagnostic specificity and reduce unnecessary treatment and patient anxiety.

Objective: This study aimed to elucidate the effect of gastric Helicobacter pylori (HP) colonization on the duodenal mucosa, focusing on intraepithelial lymphocyte (IEL) numbers and localizations.

Methods: The paired gastric and duodenal tissues from 132 patients with celiac disease (CD) and 190 individuals without CD were examined. Gastric HP status (presence and intensity) was compared with IEL counts per 100 enterocytes (IEL/100), localizations (basal-apical), and endoscopic, serologic, and clinicopathologic parameters.

Results: H pylori was detected in 176 (54.7%) gastric tissues, and its presence did not significantly change the duodenal IEL/100 counts in either CD (P = .121) or non-CD (P = .400) cases. It was seen in older individuals (P = .003), and age was also associated with HP intensity (P = .027). In non-CD cases, duodenal intraepithelial lymphocytosis (DIL) in HP-positive and HP-negative samples was 37 (33.9%) and 31 (38.3%), respectively (P = .538). Although a slight increase was observed with sparse HP colonization (+), intense colonization (+++) was significantly associated with less scalloping (P = .037), lower IEL/100 (P = .003), and antiendomysial antibody IgA (P = .048). A similar pattern was also observed in tissue transglutaminase IgA titers (P = .053).

Conclusions: Considering the effect of gastric HP on duodenal IELs, endoscopic and serologic parameters, depending on its intensity, will provide a more accurate estimation in cases where the cause of DIL is investigated.

Objective: TP53 mutations, including missense and inactivating (frameshift, splice site, and nonsense) mutations, occur in approximately 10% of myeloid neoplasms and confer adverse outcomes. Classification of myeloid neoplasms by World Health Organization and International Consensus Classification standards recognizes the importance of early detection of TP53 mutations. p53 immunohistochemistry (IHC) is a widely accessible method used to detect mutations; however, previous studies have demonstrated variable accuracy, especially for inactivating TP53 mutations. Recently, sequencing using targeted panels has seen increased use. Although highly accurate, sequencing is resource intensive and not universally available.

Methods: Using 134 bone marrow samples from patients with acute myeloid leukemia evaluated for TP53 mutation by sequencing, we assessed the concordance of p53 IHC with sequencing as well as the interrater-reliability for IHC intensity and percent positivity.

Results: Consistent with previous studies, we found that p53 IHC was strongly specific and modestly sensitive for missense mutations and that overall performance improved with dedicated hematopathology training. We also found that IHC performed poorly for inactivating mutations and was even variable between cases harboring identical amino acid changes. Low predicted transcriptional activity of p53 missense proteins correlated with a mutant pattern of IHC staining. The status of the second allele and variant allele frequency also affected the accuracy of p53 IHC as a surrogate for TP53 allele status.

Conclusion: Cases of acute myeloid leukemia with TP53 mutations predicted to have low transcriptional activity showed reduced overall survival. Our results demonstrate limited practical utility of p53 IHC for accurate evaluation of TP53 mutation status because of multifactorial confounders.

Objective: This study investigates the academic backgrounds and medical school pathology exposure among first-year pathology residents, comparing graduates from the United States and international medical schools.

Methods: A survey was administered as part of the Resident In-Service Examination First, offered by the American Society for Clinical Pathology, which assessed academic background, preparedness for residency, and prior exposure to pathology education. Associations between undergraduate pathology exposure, timing of residency selection, reported preparedness, and examination performance were analyzed.

Results: Of the 417 residents who completed the survey, 39.3% had graduated from international medical institutions. International medical graduates reported higher rates of medical school curricula that included required pathology rotations (33.5% vs 3.6%, P = .001) and greater perceived preparedness for anatomic pathology residency (28.7% vs 15.8%, P = .002), with no significant difference in examination performance. Additionally, 22.5% of US medical student respondents selected pathology before medical school, compared to only 10.4% of international medical graduates (P = .002).

Conclusions: This study highlights differences in educational exposure and perceived preparedness for pathology residency between US and international medical graduates, with international medical graduates reporting more preresidency exposure to pathology and higher perceived confidence at the start of residency. These findings suggest potential areas for curricular improvement in US medical schools to enhance pathology exposure.