Automated Ultrasensitive Detection of Light Chain Restriction in Lymphoproliferat

描述（由申请方提供）：免疫球蛋白（IG）轻链限制（LCR）检测是鉴别诊断的重要分子诊断工具，包括淋巴样增生、非典型淋巴样增生、慢性炎症和B细胞肿瘤。LCR可以通过流式细胞术（FCM）检测为细胞表面上的异常κ：λ比率。然而，FCM需要新鲜的（未固定的）组织。一些B细胞淋巴瘤缺乏FCM检测到的表面IG表达，一些显示非特异性抗体结合。通常只有福尔马林固定石蜡包埋（FFPE）组织可用于病理学评价。目前在FFPE组织中鉴定LCR的方法包括免疫组织化学（IHC）和常规显色原位杂交（CISH），其分别测量κ或λ蛋白和mRNA。然而，IHC和常规CISH都缺乏足够的灵敏度来检测许多非霍奇金淋巴瘤（NHL）中的LCR，包括最常见的NHL变体-滤泡性淋巴瘤、弥漫性大B细胞淋巴瘤、小B淋巴细胞淋巴瘤和MALT型结边缘区淋巴瘤。RNAscope是一种新型的RNA原位杂交技术，具有单分子检测灵敏度，并与FFPE组织完全兼容，使用显色或荧光检测。我们已经开发了一种手动亮视野κ和λ双链RNA ISH检测LCR，这已被证明广泛适用于所有NHL亚型。在这个项目中，我们将开发一种具有数字病理解释的全自动双重RNAscope LCR检测，这将大大促进这种新型检测的临床应用。

DESCRIPTION (provided by applicant): Immunoglobulin (Ig) light chain restriction (LCR) detection is an important molecular diagnostic tool in the differential diagnosis that includes lymphoid hyperplasia, atypical lymphoid hyperplasia, chronic inflammation, and B cell neoplasms. LCR can be detected by flow cytometry (FCM) as an abnormal kappa:lambda ratio on the cell surface. However, FCM requires fresh (unfixed) tissue. Some B cell lymphomas lack surface Ig expression detectable by FCM, some demonstrate nonspecific antibody binding. Often only formalin-fixed paraffin-embedded (FFPE) tissue is available for pathologic evaluation. Current methods of LCR identification in FFPE tissue include immunohistochemistry (IHC) and conventional chromogenic in situ hybridization (CISH), which measure kappa or lambda protein and mRNA, respectively. However, both IHC and conventional CISH lack sufficient sensitivity to detect LCR in many non-Hodgkin lymphoma (NHL), including the most common NHL variants - follicular lymphoma, diffuse large B cell lymphoma, small B lymphocytic lymphoma, and extranodal marginal zone lymphoma of MALT type. RNAscope is a novel RNA in situ hybridization technology which has single-molecule detection sensitivity and is fully compatible with FFPE tissue using either chromogenic or fluorescent detection. We have developed a manual bright-field kappa and lambda duplex RNA ISH assay for LCR, which has demonstrated broad applicability to all NHL subtypes. In this project, we will develop a fully automated duplex RNAscope LCR assay with digital pathology interpretation, which should greatly facilitate clinical adoption of this novel assay.