摘要
目的比较两种商业化二抗在免疫组织化学染色中的一致性。方法以常见肿瘤组织样本为待检组织,选取18种常用免疫组织化学染色一抗,按国际专家特设委员会的建议设立阳性对照及阴性对照。使用DAKO自动免疫组织化学染色平台,其他实验条件一致,以两种不同二抗进行免疫组织化学染色,标准组二抗为DAKO聚合物系统(DAKO EnVision FLEX,High pH),实验组二抗为Power-StainTM试剂盒(Power-StainTM 1.0 Poly HRP DAB Kit for Mouse+Rabbit)。染色后,采集图像,医师单盲法进行定位、定性以及半定量评价;两组选取同一区域,均以数字病理图像测量方法定量测量吸光度校正值、测量区域面积值以及灰度图阳性积分吸光度值,计算得到平均吸光度均值。结果两组所有样本染色的定位均准确,定性一致,两组半定量测量数据对两组强度比较、两组阳性百分率比较、两组平均吸光度均值比较,组间差异均无统计学意义。结论两种商业化二抗在免疫组织化学染色中具有良好的一致性。
Objective To compare the consistency of immunohistochemical staining between the two commercial secondary antibodies. Methods Eighteen common immunohistochemical primary antibodies were selected and positive and negative controls were set up according to the recommendations from the AD Hoc Committee of International Experts. Under the same experimental conditions, the DAKO automatic immunohistochemical staining platform was used to test two different secondary antibodies for immunohistochemical staining. The standard group for the secondary antibody was provided by the DAKO polymer system(DAKO EnVision FLEX, High pH), and the experimental group for the secondary antibody was provided by the Power-StainTM kit(Power-StainTM 1.0 Poly HRP DAB Kit for Mouse+Rabbit). Subsequently, the images were captured. A single-blind, positioning, qualitative and semi-quantitative scoring criterion was used for describing the positive stains by the experienced pathologist. Absorbance corrected values, measured area values and positive integral absorbance were detected by the digital pathology quantitative measurement in the same areas from the two groups. Then, the mean absorbance was calculated. Results The stains of all the samples from the two groups showed accurate location and consistent qualitative evaluation. No significant differences were found between the two groups in all the semi-quantitative scoring, including stain intensity, positive stain percentages and mean absorbance. Conclusion The two commercial secondary antibodies have strong consistency in the immunohistochemical staining.
作者
李曌
陈奕至
孟巍
杨智冉
林育林
张珏
孙萍萍
杜雪梅
曲丛玲
昌红
韦继学
李雁
LI Zhao;CHEN Yizhi;MENG Wei;YANG Zhiran;LIN Yulin;ZHANG Jue;SUN Pingping;DU Xuemei;QU Congling;CHANG Hong;WEI Jixue;LI Yan(Department of Pathology,Beijing Shijitan Hospital,Capital Medical University,Beijing 100038;Department of Peritoneal Cancer Surgery,Capital Medical University(Beijing Technical Training Base of Tumor Deep Hyperthermia and Whole-body Hyperthermia),Beijing 100038;Chongqing Municipal Population and Family Development Research Center,Chongqing 401120,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2021年第1期47-53,共7页
Chinese Journal of Cellular and Molecular Immunology
基金
北京市医院管理局“登峰”人才培养计划(DFL20180701)
首都临床特色应用研究与成果推广项目(Z161100000516077)
北京市优秀人才培养资助集体项目(2017400003235J007)
首都医科大学附属北京世纪坛医院重点学科建设项目(2016fmzlwk)
北京市自然科学基金(7172108)。
关键词
免疫组织化学染色技术
二抗
一致性
数字病理
immunohistochemistry
secondary antibodies
consistency
digital pathology
作者简介
李曌(1989-),女,北京人,技师,医学硕士,Tel:010-63926422,E-mail:zhaolee.carpenter@foxmail.com;通讯作者:李雁,E-mail:liyansd2@163.com。
