American journal of clinical pathology最新文献

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.

Objective: We sought to establish the US Department of Veterans Affairs (VA) Veterans Health Administration (VHA) Sequencing for Research Clinical and Epidemiology (SeqFORCE) multilaboratory consortium for SARS-CoV-2 whole-genome sequencing (WGS).

Methods: Clinical criteria were established for sending patient and employee samples from 145 VHA medical centers to 10 VHA clinical laboratories using 4 different WGS platforms. A linked pipeline among laboratories for SARS-CoV-2 clade and lineage interpretation, result transmission to electronic health records, and data storage was developed.

Results: The SeqFORCE program went live on July 1, 2021. As of December 15, 2024, 51 307 samples have been analyzed by WGS for SARS-CoV-2. The median participant age was 60 years, 76.6% were male, and 13.5% were inpatients; 96.5% were Delta, Omicron, and Recombinant sublineages; and 78.5% represented SARS-CoV-2 postvaccination samples among patients and staff.

Conclusions: Establishment of VA SeqFORCE enabled national population analysis for use in epidemiologic response and population health policy as well as expanded SARS-CoV-2 sequencing capacity to meet the demand for clinical and public health sequencing. The program consolidated operations using standardized procedures, test setup, analysis, reporting, and tracking. It also improved oversight and governance of VA contributions to global databases, mitigated system inefficiencies, and prepared VHA for future genomic challenges.

Objective: Cytologic examination, which confirms the presence or absence of malignant cells, detects malignant cells from various organs, with adenocarcinoma as the most common histologic type. We developed a deep learning model to detect malignant cells in images obtained following effusion cytology.

Methods: The deep learning model was created using the YOLOv8 object detection algorithm (Roboflow, Inc) and 275 cases of adenocarcinoma comprising 12 182 images and 29 245 labels as well as 188 cases negative for malignancy comprising 1980 images.

Results: The adenocarcinoma test dataset exhibited Precision, Recall, F1, and mean average Precision scores of 0.909, 0.911, 0.910, and 0.955, respectively. The number of adenocarcinoma test images in which 1 or more malignant cells were detected was 2710 of 2731. The sensitivity in the nonadenocarcinoma dataset was 97.1%, and the false-positive rate in the negative-for-malignancy dataset was 7.3%. The accuracy, sensitivity, and specificity of the model using all the test datasets were 96.3%, 98.5%, and 92.7%, respectively.

Conclusions: Although some issues regarding cell annotation when creating an object detection model remain, the accuracy is sufficient to assist cancer screening in effusion cytology. It is vital to reliably detect malignant cells in effusion cytology, and the further development of automated systems to reduce false-negative results is expected.

Objective: Rare large B-cell lymphomas (LBCLs) present with concurrent or subsequent lymphomatous effusion (solid-effusion LBCL, SE-LBCL), which may have an inferior prognosis compared with their noneffusion counterpart. In addition, the relationship between SE-LBCL and human herpesvirus 8-negative effusion-based LBCL (EB-LBCL) remains unclear.

Methods: We collected 141 cases of SE-LBCL and a control cohort of 101 cases of stage IV solid-only LBCL. The clinicopathologic features were analyzed and compared between SE-LBCL and solid-only LBCL.

Results: Patients with SE-LBCL had a median age of 67 years with a male-to-female ratio of 1.3:1. Eighty-six patients had concurrent solid lymphoma and lymphomatous effusion, whereas 55 developed lymphomatous effusion subsequently. Most cases involved the pleural cavities (79%, 112/141), followed by the peritoneal (21%, 29/141) and pericardial (11%, 16/141) cavities. BCL6, CD10, and MUM1 were expressed in 77% (90/117), 46% (60/130), and 61% (58/95) of cases, respectively, and 58% (71/123) were subclassified into the germinal center B-cell (GCB) subtype. Rearrangements of BCL2, BCL6, and MYC were detected in 42% (31/73), 35% (22/63), and 40% (35/88), respectively, and 22% (19/87) had both MYC and BCL2 rearrangements. The patients with SE-LBCL had a dismal prognosis, with a median survival of 5.7 months, which was significantly worse than solid-only LBCL (147.5 months; P < .0001).

Conclusions: The pathologic features of SE-LBCL were similar to those of solid-only LBCL but distinct from those of EB-LBCL; in particular, lymphomatous effusion was an independently adverse prognostic factor in LBCL. Our study underscores the need for surveillance of lymphomatous effusion during LBCL staging and development of effective therapeutic regimens for SE-LBCL.