当前分析免疫组织化学的方法是劳动密集型的,并且经常被观察者之间的变异性所混淆。当在较大样品中鉴定小的临床重要组群时,分析是耗时的。这项研究训练了QuPath,一个开源的图像分析程序,从包含正常结肠和IBD-CRC的组织微阵列中准确鉴定MLH1缺陷的炎症性肠病相关结直肠癌(IBD-CRC)。组织微阵列(n=162个核心)进行MLH1免疫染色,数字化,并导入到QuPath中。小样本(n=14)用于训练QuPath以检测阳性与无MLH1和组织组织学(正常上皮,肿瘤,免疫浸润物,基质)。将该算法应用于组织微阵列,并在大多数有效病例(73/99,73.74%)中正确识别组织组织学和MLH1表达,在一种情况下,错误地识别了MLH1状态(1.01%),并标记25/99(25.25%)例进行人工审查。定性审查发现了标记核心的五个原因:组织数量少,多样/非典型形态,过度的炎症/免疫浸润,正常粘膜,或弱/斑片状免疫染色。在分类核心(n=74)中,QuPath对MLH1缺陷性IBD-CRC的鉴别为100%(95%CI80.49,100)敏感和98.25%(95%CI90.61,99.96)特异性;κ=0.963(95%CI0.890,1.036)(p<0.001)。该过程可以在诊断实验室中有效地自动化,以检查所有结肠组织和肿瘤的MLH1表达。
Current methods for analysing immunohistochemistry are labour-intensive and often confounded by inter-observer variability. Analysis is time consuming when identifying small clinically important cohorts within larger samples. This study trained
QuPath, an open-source image analysis program, to accurately identify MLH1-deficient inflammatory bowel disease-associated colorectal cancers (IBD-CRC) from a tissue microarray containing normal colon and IBD-CRC. The tissue microarray (n = 162 cores) was immunostained for MLH1, digitalised, and imported into
QuPath. A small sample (n = 14) was used to train
QuPath to detect positive versus no MLH1 and tissue histology (normal epithelium, tumour, immune infiltrates, stroma). This algorithm was applied to the tissue microarray and correctly identified tissue histology and MLH1 expression in the majority of valid cases (73/99, 73.74%), incorrectly identified MLH1 status in one case (1.01%), and flagged 25/99 (25.25%) cases for manual review. Qualitative review found five reasons for flagged cores: small quantity of tissue, diverse/atypical morphology, excessive inflammatory/immune infiltrations, normal mucosa, or weak/patchy immunostaining. Of classified cores (n = 74),
QuPath was 100% (95% CI 80.49, 100) sensitive and 98.25% (95% CI 90.61, 99.96) specific for identifying MLH1-deficient IBD-CRC; κ = 0.963 (95% CI 0.890, 1.036) (p < 0.001). This process could be efficiently automated in diagnostic laboratories to examine all colonic tissue and tumours for MLH1 expression.