American journal of clinical pathology最新文献

Objective: We sought to present an alternative liquid-based cytologic (aLBC) and an agarose-based cell block (CB) preparation method for formalin-fixed fine-needle aspirations (FNAs), using breast cancer as a model, and to perform a validation procedure for immunohistochemical (IHC) assays.

Methods: Between 2024 and 2025, 18 breast cancer FNA cases were collected and processed into agarose-based CB, and by applying the dropped reaspirated monolayer dried preparation method, into aLBC. Matched formalin-fixed, paraffin-embedded surgical specimens from the same patients served as gold-standard controls. Ten immunomarkers were assessed using validated IHC protocols. Human epidermal growth factor receptor 2 (HER2) status was evaluated by both IHC and dual-probe fluorescence in situ hybridization (FISH) according to 2023 American Society of Clinical Oncology/College of American Pathologists guidelines. Staining intensity, percentage of positive cells, and marker concordance were analyzed across all specimen types. Diagnostic performance metrics were calculated, and intermethod agreement was assessed using Cohen's κ coefficient (κ > 0.75 considered acceptable).

Results: The sensitivity, specificity, and either negative or positive predictive values and the accuracy values were 100% at all the tested immunostains and at the HER2 FISH assays.

Conclusions: Accurate diagnosis from scant FNA material is an increasing demand in cytopathology. The validated aLBC and CB preparation methods proved to be cost-effective, efficient for ancillary testing, and reliable for IHC, even in low-cellularity samples.

Objective: The Hologic Genius Digital Diagnostics System (HGDDS) analyzes ThinPrep Papanicolaou (Pap) tests (TPPTs) to assist in detecting cervical lesions. The aim of this study was to determine the sensitivity of the HGDDS in identifying commonly diagnosed microorganisms in Pap tests.

Methods: A total of 305 TPPT cases were selected from Magee Women's Hospital, University of Pittsburgh, consisting of 244 cases with microorganism diagnoses (a total of 262 cases of Actinomyces, Candida spp, herpes simplex virus [HSV], and Trichomonas) and 61 cases without microorganisms. Slides were scanned and then subjected to artificial intelligence (AI) analysis using the HGDDS and subsequently reviewed on a digital workstation by a cytologist, followed by a resident and a cytopathologist who made the final diagnoses.

Results: Diagnosis using the HGDDS demonstrated high sensitivity across all microorganisms (95.4%). Herpes simplex virus detection was comparatively lower (82.5%). Of the microorganisms, 85.2% were displayed in the first gallery of 30 images within row 5, 7.2% presented in the first gallery outside of row 5, and 7.6% presented in the hidden gallery of images. Among the 12 cases with missed diagnoses, 3 of 5 Candida spp and 3 of 7 HSV organisms were not presented within the 60 images selected by HGDDS. In another 6 cases, microorganisms were found within the 60 fields, but none were present in row 5.

Conclusions: Very high sensitivity was observed for TPPTs across 3 of 4 common microorganisms on the HGDDS, although sensitivity was relatively lower for detecting HSV. Understanding morphologic patterns of various microorganisms in detection misses by the HGDDS may help guide the implementation of AI-assisted cervical cancer screening systems.

Objective: Our objective was to explore how variations in the number and distribution of tested concentrations around the presumed lower limit of detection (LLOD) affect the estimate and its confidence interval.

Methods: We reviewed published LLOD evaluations and conducted sensitivity analyses to assess how limiting the number and distribution of input concentrations affects LLOD estimates. We systematically reduced the number of tested concentrations and varied their distribution, either centered or top-weighted around the LLOD, to evaluate impacts on the estimate and confidence interval.

Results: When data sets are restricted but remain centered, the estimated LLOD lowers. When data sets are restricted to top-weighted concentrations, the estimated LLOD lowers, and the confidence intervals widen considerably. In all cases of data restriction, model fit, as measured by the Akaike information criterion, suffers, the effects of which are most severe in the top-weighted scenarios.

Conclusions: These findings reinforce recommendations from the Clinical and Laboratory Standards Institute and highlight the need for caution when constrained testing designs are used in LLOD estimation.

Objective: Default settings in electronic medical records (EMRs) can drive overordering of diagnostic tests. After transitioning to Epic, our hospital network noted a sharp rise in acid-fast bacilli (AFB) and fungal culture orders from operating rooms (ORs), likely due to 1-click ordering tools ("quick buttons") in the Epic OpTime module. We assessed the impact of removing these buttons on test utilization.

Methods: This multicenter study included 8 medical centers and compared two 9-month periods: preintervention (January to September 2023) and postintervention (December 2023 to August 2024). The AFB and fungal cultures remained orderable via manual entry. Order volumes, positivity rates, and turnaround times (TATs) were extracted from the Laboratory Information System.

Results: Postintervention, AFB orders decreased by nearly 50% (P < .001) while positivity doubled (1.74% to 3.64%, P < .001). Median AFB TAT improved by more than 2 hours (P < .001). Fungal culture orders declined 35% (P < .001), with no significant change in positivity (6.37% vs 6.25%, P = .83). Median fungal TAT improved by more than 1 hour (P < .001).

Conclusions: Removing OR quick buttons significantly reduced unnecessary AFB and fungal culture orders and improved TAT without reducing detection of clinically important infections. This is the first study to evaluate the impact of EMR quick buttons on -microbiology utilization.

Objective: Using a multidisciplinary approach, we sought to develop effective information technology (IT)-driven solutions to reduce both incoming and outgoing telephone communication to minimize errors and complaints while improving efficiency and workplace satisfaction.

Methods: We built 2 IT-driven workflows, one targeting incoming phone calls and the other outgoing calls. To reduce incoming calls, we implemented a shell test with a monitored dashboard, enabling nursing staff to electronically request blood product release for their patients. To reduce outgoing calls, we developed a web-based application for critical values that sends a secure message with patient information and the critical value, which the provider must acknowledge or refuse. Critical value disposition information is then fed back into the critical value application, and in the event of refusal to acknowledge, the technologist initiates timely escalation via manual workflows. We assessed incoming call volume for blood products, formal complaints, critical value notification turnaround time, and end-user satisfaction.

Results: We significantly reduced incoming calls and formal complaints and received positive end-user satisfaction feedback for the critical value application. We did not find a difference in critical value reporting turnaround time after the intervention.

Conclusions: We successfully built IT-driven solutions, which reduced incoming and outgoing phone calls in our clinical laboratories. These efforts reduced complaints and created systems well received by end users.

Objective: BRAF V600E/K mutations are significant drivers of metastatic melanoma, and therapies that selectively target these mutations have significantly improved patient long-term survival. Emerging evidence suggests that tumors harboring BRAF non-V600E/K mutations are susceptible to MEK-targeting therapies, but these mutations are not always assessed in clinical workup. To assess whether patient stratification for targeted therapy can be improved, we characterized the prevalence and nature of BRAF non-V600E/K mutations in a cohort of patients diagnosed with metastatic melanoma.

Methods: We extracted DNA from 99 specimens collected between June 2019 and September 2021. The samples were analyzed using next-generation sequencing to identify mutations in BRAF exon 15 and to assess their susceptibility to targeted therapies.

Results: Eleven percent of patient specimens contained BRAF non-V600E/K mutations in exon 15. Of the mutations identified, all have prior evidence of susceptibility to targeted MEK therapies. Among those patients, less than half received targeted MEK therapies.

Conclusions: Next-generation sequencing is a sensitive platform for profiling BRAF exon 15 mutations that cannot be detected by other techniques. Our results support the routine sequencing of BRAF exon 15 to improve patient stratification for therapy.

Objective: To describe the prevalence and clinicopathologic associations of FGFR-altered urinary tract carcinomas in institutions incorporating reflex testing.

Methods: Next-generation sequencing was prospectively performed on urinary tract carcinomas for the detection of FGFR1-4 alterations at 2 tertiary care centers (2021-2025), using the Oncomine Comprehensive Assay (OCA) v3 DNA and OCA Plus RNA. Reflex testing was conducted on metastatic and/or advanced (pT3/4) carcinomas.

Results: The cohort included 366 patients (239 lower tract carcinomas, 72 upper tract carcinomas, and 55 metastases). Median age was 72.5 years (range, 36-97). Fifty-nine (16.1%) tumors were FGFR-altered. Forty-nine (13.4%) patients with actionable FGFR alterations (33 FGFR3 mutations, 13 FGFR3 fusions, and 3 FGFR2 mutations) were all 55 years or older (P = .097). The prevalence of actionable FGFR alterations was significantly higher in upper vs lower tract carcinomas (23.8% vs 13.8%, P = .007) and in lung metastases vs other metastatic sites (57.1% vs 10.4%, P = .002). A higher frequency was also seen in metastases vs primary tumors (16.4% vs 12.9%), although this difference did not reach statistical significance (P = .482). Actionable FGFR alterations were observed in urothelial carcinoma not otherwise specified (40/261) and in urothelial carcinoma with squamous differentiation (6/43), micropapillary features (2/11), or nested features (2/7).

Conclusions: The detection rate for FGFR1-4 alterations in a real-world, dual-institution cohort of urinary tract carcinomas was reported.

Objective: Gastrointestinal (GI) symptoms are common in systemic mastocytosis (SM). We hereby report the clinicopathologic and molecular features of GI mastocytosis.

Methods: The study includes 104 patients with suspected SM. Of 258 GI biopsy -specimens, 33 (20 patients, 19%) were mastocytosis-positive, with 19 (16 patients) initially missed.

Results: Tryptase (n = 29) was weakly positive/negative in two-thirds of the mastocytosis-positive GI biopsy specimens. Additional immunohistochemistry (n = 23) showed positive mast cell expression for CD30 (n = 2), CD123 (n = 7), and PD-L1 (n = 12), as well as increased PD-1-positive cells (n = 4). Sixty patients had positive bone marrow (BM) for SM (SM-BM+), including 19 with positive GI biopsy specimens (SM-GI+). Almost all (43/44) SM-BM- patients were SM-GI-. The SM-GI vs SM-BM status showed high positive predictive value (95%) and specificity (98%). Patients with SM with mastocytosis-positive vs mastocytosis-negative GI biopsy specimens had higher serum tryptase levels (P = .04). KIT D816V mutation was detected in 7 of 13 and 12 of 13 mastocytosis-positive GI biopsy specimens using allele-specific polymerase chain reaction and droplet digital polymerase chain reaction (ddPCR), respectively.

Conclusions: GI mastocytosis is highly predictive of SM in BM; however, most SM-BM+ patients are SM-GI-. The SM-BM- patients are very unlikely to be SM-GI+. Because GI neoplastic mast cells are often tryptase negative, diagnostic evaluation should include CD117 and CD25 immunohistochemistry, as well as KIT D816V mutation analysis by ddPCR as needed.

Objective: To develop and validate a machine learning model for predicting oligoclonal band (OCB) positivity using routine cerebrospinal fluid (CSF) and serum biochemical markers to improve the diagnostic accuracy and efficiency of assessing intrathecal immunoglobulin G (IgG) synthesis.

Methods: In this retrospective study (n = 1709), an ensemble model was developed using 8 refined CSF and serum parameters. Combining optimized CatBoost, XGBoost, and LightGBM classifiers, the model was trained and evaluated using a 2-phase workflow, including 5-fold cross-validation and validation on independent internal (n = 342) and external (n = 49) cohorts.

Results: The developed ensemble model achieved a receiver operating characteristic-area under the curve (ROC-AUC) of 0.902 on the internal test set, significantly outperforming the conventional IgG index (ROC-AUC, 0.795). At its optimal threshold, the model demonstrated an accuracy of 0.830, with a sensitivity of 0.714 and a specificity of 0.916. On the external validation cohort, it achieved 90% accuracy and 96% sensitivity.

Conclusions: A novel machine learning ensemble model accurately predicts OCB positivity using routine laboratory data and demonstrates superior performance compared with the IgG index. This approach represents a significant step in applying artificial intelligence in laboratory medicine, with the potential to enhance diagnostic efficiency. Prospective, multicenter validation is essential for broader clinical implementation.

Objective: The OrganOx metra is a recent innovation approved by the US Food and Drug Administration that enables continuous ex vivo perfusion of a donor liver prior to transplantation. A mixture of human blood products, added nutrients, and drugs is perfused throughout the liver to sustain its viability. This study determines whether the perfusate matrix affects the analytical accuracy of biochemical parameters used to evaluate the function of the donor liver before transplantation.

Methods: A mixing study was conducted to evaluate the percent recovery of total bilirubin, aspartate aminotransferase, alanine aminotransferase, lactic acid, glucose, and lactate dehydrogenase in OrganOx metra perfusate using the Beckman Coulter AU5800 automated platform. Perfusate pools were mixed with pooled patient plasma (Li Hep) in the following ratios: 1:4, 1:1, and 4:1. These mixtures were measured alongside straight patient plasma and straight perfusate.

Results: Agreement between expected and measured values, with a percent recovery for all tested analytes, was between 92% and 109%. Detrimental matrix effects were not observed.

Conclusions: The Beckman Coulter AU5800 analyzer provides reliable and accurate biochemical analysis of OrganOx metra perfusate during ex vivo preservation. The findings validate its suitability for perfusate analysis, supporting its potential use in clinical and research settings.