Investigating Various Thresholds as Immunohistochemistry Cutoffs for Observer Agreement

Ali, Asif; Bell, Sarah; Bilsland, Alan; Slavin, Jill; Lynch, V.; Elgoweini, Maha; Derakhshan, Mohammad H.; Jamieson, Nigel B.; Chang, David C.; Brown, Victoria; Denley, Simon M.; Orange, Clare; McKay, Colin; Carter, Ross; Oien, Karin A.; Duthie, Fraser

doi:10.1097/pai.0000000000000357

Cited by 7 publications

(7 citation statements)

References 52 publications

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“…Our study shows that non-pathologists are able to generate reproducible IHC scores after appropriate training, which is in line with previous studies (17)(18)(19)40). Interestingly, intraobserver kappa values of non-pathologists in the current study were similar to those previously reported for pathologists (23,43). In general, across all three markers, disagreements were limited to one-category discordances (e.g., <10% vs. 11%-50%) for all comparisons.…”

Section: Intraobserver Agreement Of Non-pathologistssupporting

confidence: 92%

Validity and Reproducibility of Immunohistochemical Scoring by Trained Non-Pathologists on Tissue Microarrays

Jenniskens

Offermans

Samarska

et al. 2021

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Background: Scoring of immunohistochemistry (IHC) staining is often done by non-pathologists, especially in large-scale tissue microarray (TMA)-based studies. Studies on the validity and reproducibility of scoring results from non-pathologists are limited. Therefore, our main aim was to assess interobserver agreement between trained non-pathologists and an experienced histopathologist for three IHC markers with different subcellular localization (nucleus/membrane/cytoplasm). Methods: Three non-pathologists were trained in recognizing adenocarcinoma and IHC scoring by a senior histopathologist. Kappa statistics were used to analyze interobserver and intraobserver agreement for 6,249 TMA cores from a colorectal cancer series. Results: Interobserver agreement between non-pathologists (independently scored) and the histopathologist was “substantial” for nuclear and membranous IHC markers (κrange = 0.67–0.75 and κrange = 0.61–0.69, respectively), and “moderate” for the cytoplasmic IHC marker (κrange = 0.43–0.57). Scores of the three non-pathologists were also combined into a “combination score” (if at least two non-pathologists independently assigned the same score to a core, this was the combination score). This increased agreement with the pathologist (κnuclear = 0.74; κmembranous = 0.73; κcytopasmic = 0.57). Interobserver agreement between non-pathologists was “substantial” (κnuclear = 0.78; κmembranous = 0.72; κcytopasmic = 0.61). Intraobserver agreement of non-pathologists was “substantial” to “almost perfect” (κnuclear,range = 0.83–0.87; κmembranous,range = 0.75–0.82; κcytopasmic = 0.69). Overall, agreement was lowest for the cytoplasmic IHC marker. Conclusions: This study shows that adequately trained non-pathologists are able to generate reproducible IHC scoring results, that are similar to those of an experienced histopathologist. A combination score of at least two non-pathologists yielded optimal results. Impact: Non-pathologists can generate reproducible IHC results after appropriate training, making analyses of large-scale molecular pathological epidemiology studies feasible within an acceptable time frame.

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Section: Intraobserver Agreement Of Non-pathologistssupporting

confidence: 92%

Validity and Reproducibility of Immunohistochemical Scoring by Trained Non-Pathologists on Tissue Microarrays

Jenniskens

Offermans

Samarska

et al. 2021

Cancer Epidemiology, Biomarkers &Amp; Prevention

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“…2 The immunohistochemical positivity for IDH1 was considered strong with 3+ positivity and weak with 1-2+ positivity. 5,7 All the cases have been blindly examined by an expert neuropathologist and a general pathologist together. It means that there has been no information about the final diagnosis when the IHC slides were being reviewed by the pathologist.…”

Section: Methodsmentioning

confidence: 99%

“…The slides were reviewed and the best representative block was selected for IHC with mouse monoclonal anti-R132H-IDH1 antibody (H09, Dianova, dilution 1:50, citrate-EDTA for antigen retrieval) and P53 antibody (Dako, clone DO-7, dilution 1:100, citrate-EDTA [Ethylenediamine tetraacetic acid] for antigen retrieval). Cytoplasmic positivity for IDH1 1 , 5 and nuclear positivity for P53 2 were considered positive. We considered every positive case as positive and every negative P53 as negative.…”

Section: Methodsmentioning

confidence: 99%

“…4 Majority of IDH1 mutations occur in exon 4 at codon 132, where a transition changes a single amino acid from arginine to histidine (R132H). 5 It has been found in the glial cells especially astrocytoma World Health Organization (WHO) grade II with a remarkably high frequency (around 75%), and IDH mutation has become definitional for infiltrating gliomas in adults. 4,6 In this study, we are trying to evaluate the role of IDH1 mutation in combination with P53 mutation by immunohistochemistry (IHC) method to differentiate reactive gliosis from 2 Clinical Pathology low-grade astrocytoma (grade II).…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Simple Panel of IDH1 and P53 in Differential Diagnosis Between Low-Grade Astrocytoma and Reactive Gliosis

Geramizadeh

Kohandel-Shirazi

Soltani

2021

Clin Med�Insights�Pathol

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Background: Reactive gliosis is a response of glial tissue to different types of injury such as brain abscess, trauma, hemorrhage, or even neoplastic process. In some circumstances, especially when the tissue biopsy is small, there may be difficulty to discriminate this reactive condition with low-grade diffuse astrocytoma (World Health Organization [WHO] grade II) by conventional hematoxylin and eosin (H&E) slides, so some immunohistochemical and molecular markers have been introduced for this differential diagnosis. One of the important aspects of updated WHO classification in 2016 has been dividing some of the glial tumor according to IDH1 (isocitrate dehydrogenase 1) mutation. Objectives: In this study, we tried to evaluate IDH1 and P53 mutation by immunohistochemistry as a simple and highly specific and sensitive method to differentiate low-grade astrocytoma and reactive gliosis. Material and methods: For 5 years (2013-2018), 50 cases of clinically documented reactive gliosis and 50 cases of low-grade astrocytoma were evaluated for the presence or absence of IDH1 and P53 mutation by immunohistochemistry. Results: Isocitrate dehydrogenase 1 was positive in 92% and 4% of the astrocytoma and reactive gliosis cases and P53 was positive in 90% and 4% of the cases with the final diagnosis of astrocytoma and reactive gliosis, respectively. Discussion and conclusion: Combination of P53 and IDH1 as an immunohistochemical panel showed specificity of 96% and sensitivity of 91% for differential diagnosis of reactive gliosis and low-grade astrocytoma. These 2 markers can be extremely helpful for this differential diagnosis.

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Immunohistochemistry Quality Management and Regulation

Prichard

2022

Handbook of Practical Immunohistochemistry

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Investigating Various Thresholds as Immunohistochemistry Cutoffs for Observer Agreement

Abstract: Background

Cited by 7 publications

References 52 publications

Validity and Reproducibility of Immunohistochemical Scoring by Trained Non-Pathologists on Tissue Microarrays

Validity and Reproducibility of Immunohistochemical Scoring by Trained Non-Pathologists on Tissue Microarrays

A Simple Panel of IDH1 and P53 in Differential Diagnosis Between Low-Grade Astrocytoma and Reactive Gliosis

Immunohistochemistry Quality Management and Regulation

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