利用固有形态学截断重新发明核组织评分:蓝棕色h评分(BBC-HS)。

IF 1.3 4区 医学 Q3 ANATOMY & MORPHOLOGY
Phillipe Price, Usharani Ganugapati, Zoran Gatalica, Archan Kakadekar, James Macpherson, Louise Quenneville, Henrike Rees, Elzbieta Slodkowska, Janarthanee Suresh, Darryl Yu, Hyun J Lim, Emina E Torlakovic
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引用次数: 0

摘要

免疫组织化学(IHC)是一种广泛用于大量诊断、预后和预测生物标志物的测试方法。虽然IHC是一种定性方法,除了基于阈值的分层(阳性与阴性)之外,其中一些生物标志物表达水平的增加通常会导致更强烈的染色,这与特定的诊断、预后和治疗反应有关。至关重要的是,阳性和阴性染色之间的描述性阈值,以及经常使用的染色强度分级类别(例如,1+,2+,3+)之间的阈值是标准化和可重复的。历史分数(H-score)是一种常用的评分系统,它利用了这些类别。我们的研究介绍了基于苏木精和二氨基联苯胺(DAB)之间的颜色相互作用的核IHC生物标志物检测的阳性和阴性结果的截止点分类和染色强度的分级分类;蓝棕色h值(BBC-HS)。对6例弥漫性大b细胞淋巴瘤进行核标记物MUM1染色。12名读卡器进行h -评分。为读者提供了简短的教程和插图说明。本研究新颖的评分系统利用DAB (DAB,棕色染色)和苏木精(蓝色反染色)之间的相互作用,在“0”(阴性细胞核)、“1+”(弱阳性细胞核)、“2+”(中度阳性细胞核)和“3+”(强阳性细胞核)之间设置阈值。阅读者记录了300个细胞的分数。计算Krippendorff alpha (K-alpha)和类内相关系数(ICC)。我们还评估了在使用K-alpha和ICC计数前100个细胞、前200个细胞和总共300个细胞时,可靠性是否得到改善。为了评估每位读者的表现,计算每种情况下的平均h分和阳性分数百分比(PPS),并计算每位读者的得分与平均值之间的偏差。H-score的K-alpha为0.86,PPS的K-alpha为0.76。H-score的ICC为0.96,PPS为0.92。H-score的偏倚范围为-58 ~ 41,而PPS的偏倚范围为-27% ~ 33%。总的来说,大多数读者的偏见都很低。与平均值相比,两名读者一直在划线,两名读者一直在划线。对于核免疫组化生物标志物检测,我们新提出的截止点提供了高度可靠/可重复的阳性和阴性结果阅读器和使用现有形态学参数的染色强度分级分类。BBC-HS易于教学,适用于人眼和图像分析。BBC-HS的应用将有助于开发新的可靠/可重复的IHC生物标志物评分方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Reinventing Nuclear Histo-score Utilizing Inherent Morphologic Cutoffs: Blue-brown Color H-score (BBC-HS).

Reinventing Nuclear Histo-score Utilizing Inherent Morphologic Cutoffs: Blue-brown Color H-score (BBC-HS).

Reinventing Nuclear Histo-score Utilizing Inherent Morphologic Cutoffs: Blue-brown Color H-score (BBC-HS).

Reinventing Nuclear Histo-score Utilizing Inherent Morphologic Cutoffs: Blue-brown Color H-score (BBC-HS).

Immunohistochemistry (IHC) is a testing methodology that is widely used for large number of diagnostic, prognostic, and predictive biomarkers. Although IHC is a qualitative methodology, in addition to threshold-based stratification (positive vs. negative), the increasing levels of expression of some of these biomarkers often lead to more intense staining, which published evidence linked to specific diagnosis, prognosis, and responses to therapy. It is essential that the descriptive thresholds between positive and negative staining, as well as between frequently used graded categories of staining intensity (eg, 1+, 2+, 3+) are standardized and reproducible. Histo-score (H-score) is a frequently used scoring system that utilizes these categories. Our study introduces categorization of the cutoff points between positive and negative results and graded categories of staining intensity for nuclear IHC biomarker assays based on color interaction between hematoxylin and diaminobenzidine (DAB); the Blue-brown Color H-score (BBC-HS). Six cases of diffuse large B-cell lymphoma were stained for a nuclear marker MUM1. The staining was assessed by H-score by 12 readers. Short tutorial and illustrated instructions were provided to readers. The novel scoring system in this study uses the interaction between DAB (DAB, brown stain) and hematoxylin (blue counterstain) to set thresholds between "0" (negative nuclei), "1+" (weakly positive nuclei), "2+" (moderately positive nuclei), and "3+" (strongly positive nuclei). The readers recorded scores for 300 cells. Krippendorff alpha (K-alpha) and intraclass correlation coefficient (ICC) were calculated. We have also assessed if reliability improved when counting the first 100 cells, first 200 cells, and for the total 300 cells using K-alpha and ICC. To assess the performance of each individual reader, the mean H-score and percent positive score (PPS) for each case was calculated, and the bias was calculated between each reader's score and the mean. K-alpha was 0.86 for H-score and 0.76 for PPS. ICC was 0.96 for H-score and 0.92 for PPS. The biases for H-score ranged from -58 to 41, whereas for PPS it ranged from -27% to 33%. Overall, most readers showed very low bias. Two readers were consistently underscoring and 2 were consistently overscoring compared with the mean. For nuclear IHC biomarker assays, our newly proposed cutoffs provide highly reliable/reproducible results between readers for positive and negative results and graded categories of staining intensity using existing morphologic parameters. BBC-HS is easy to teach and is applicable to both human eye and image analysis. BBC-HS application should facilitate the development of new reliable/reproducible scoring schemes for IHC biomarkers.

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来源期刊
Applied Immunohistochemistry & Molecular Morphology
Applied Immunohistochemistry & Molecular Morphology ANATOMY & MORPHOLOGY-MEDICAL LABORATORY TECHNOLOGY
CiteScore
3.20
自引率
0.00%
发文量
153
期刊介绍: ​Applied Immunohistochemistry & Molecular Morphology covers newly developed identification and detection technologies, and their applications in research and diagnosis for the applied immunohistochemist & molecular Morphologist. Official Journal of the International Society for Immunohistochemisty and Molecular Morphology​.
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