RELIABILITY OF EBV DETECTION IN HODGKINS DISEASE TUMORS

描述（申请人描述）本拟定R 03研究将是 首次检查EBER原位杂交（ISH）的观察者可靠性 和LMP-1免疫组织化学（IHC），形态学分子检测广泛 在临床研究中用于检测EB病毒（EBV）潜伏感染 肿瘤细胞。 由于这些测定法易于使用、快速且适用于 存档的肿瘤标本，它们非常适合大规模的分子生物学研究。 流行病学项目。 霍奇金病（HD）是一种常见的年轻人癌症， EBER ISH和LMP-1 IHC在大约40%的成年人中检测到EBV。 病例，并且病毒定义之间存在显著的流行病学差异， 然而，即使在治疗后，HD研究中的EBV患病率也不同。 人口流行病学差异的统计控制。 问题 与观察员的可靠性可能是部分负责;然而，虽然分析 解释是主观的，可靠性从未被评估过。 或者测试。 本项目将确定评分者间和评分者内的可靠性 EBER ISH和LMP-1 IHC检测在40例HD肿瘤中的EBV检测，基于 由四名血液病理学家分别对切片进行两次审查， 两个实验室 从先前项目评估的肿瘤标本中，我们将 随机选取EBV阳性和阴性病例各10例 最常见的组织学亚型，以最大限度地提高Kappa值的精度 统计数据。 从这40个库存标本中，每个实验室将对40个载玻片进行染色 EBER ISH为40，LMP-1 IHC为40，每次测定产生80个载玻片。 在NCCC， 两个实验室的染色玻片将在检测试剂盒内混合，重新标记 给盲评者机构身份证明，并分批邮寄给 评估者，记录他们的解释（EBV阳性，EBV阴性） 并对解释难度进行排序和描述。 第一次和第二次 评审时，工作人员将对幻灯片重新编号，使评分者不知道评审顺序。 每次测定将进行640次评级（320次第一和320次第二）。 与 这些数据，我们将计算多个评分员的Kappa统计数据， 确定评估者间和评估者内的一致性， 比色法 还将比较Kappas在 每个实验室使用相同的标本，以检测实验室对判读的影响 准确性，以及两种组织学亚型检测差异， 导致亚型间EBV流行率变化。 将制定调查结果 具体的解释准则。 研究优势包括： 第一次审查两个共同的评级者之间和内部的一致性 使用具有流行病学潜力的临床EBV检测试验 在研究EBV相关癌症中的应用; 解释指南;使用现有的病理学材料 效率和组装一个案件系列，最大限度地提高精度; 样本量足以估计组织学间差异， 统计功效;与EBV检测领域的专家合作; 并关注HD，一种常见的EBV相关癌症， EBV分类的临床意义。

DESCRIPTION (Applicant's Description) This proposed R03 study will be the first to examine observer reliability of EBER in situ hybridization (ISH) and LMP-1 immunohistochemistry (IHC), morphologic molecular assays widely used in clinical studies to detect Epstein-Barr virus (EBV) latent infection of tumor cells. As these assays are easy to use, rapid, and applicable to archived tumor specimens, they are ideally suited for large-scale molecular epidemiologic projects. In Hodgkin's disease (HD), a common cancer of young adults, EBER ISH and LMP-1 IHC have detected EBV in about 40 percent of cases, and marked epidemiologic differences exist between the virus-defined subgroups, However, EBV prevalence varies among HD studies even after statistical control for epidemiologic differences in populations. Problems with observer reliability may be partly responsible; yet, although assay interpretation is subjective, reliability has never been evaluated for either test. This project will determine inter- and intra-rater reliability of EBER ISH and LMP-1 IHC assays in EBV detection in 40 HD tumors, based on two reviews by each of four hematopathologists of slides, prepared at each of two labs. From tumor specimens evaluated for prior projects, we will randomly select 10 EBV-positive and 10 EBV-negative cases of each of the two most common histology subtypes, to maximize precision of the kappa statistic. From these 40 banked specimens, each lab will stain 40 slides for EBER ISH and 40 for LMP-1 IHC, yielding 80 slides per assay. At NCCC, stained slides from the two labs will be intermixed within assays, relabeled to blind raters to institutional identification, and mailed in batches to raters, who will record their interpretations (EBV-positive, EBV-negative) and both rank and describe interpretive difficulty. Been lst and 2nd reviews, staff will renumber slides to blind raters to the review sequence. 640 ratings (320 first and 320 second) will occur for each assay. With these data, we will compute kappa statistics for multiple raters to determine inter- and intra-rater agreement occurring beyond chance for each assay. Kappas will also be compared for agreement on slides prepared on the same specimens by each lab to detect any lab effect on interpretation accuracy, and for the two histologic subtypes to detect differences that may lead to variation in EBV prevalence by subtype. Findings will be developed into specific interpretation guidelines. Study strengths include: the first examination of both inter- and intra-rater agreement for two commonly used clinical EBV detection assays with potential for epidemiologic application in studying EBV-linked cancer; development of explicit interpretation guidelines; use of existing pathology materials for efficiency and for assembling a case series which maximizes precision; adequate sample size to estimate inter-histology differences with sufficient statistical power; collaboration with experts in the field of EBV detection; and focus on HD, a common EBV-related cancer with both epidemiologic and clinical importance for EBV classification.