Automated Ultrasensitive Detection of Light Chain Restriction in Lymphoproliferat

淋巴细胞增殖中轻链限制的自动超灵敏检测

• 批准号: 8935761
• 负责人: Xiao-Jun Ma
• 金额: $ 52.95万
• 依托单位: ADVANCED CELL DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-14 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8935761
• 关键词: Address; Adoption; Antibodies; Aspirate substance; Atypical Lymphoproliferative Disorder; Automation; B lymphoid malignancy; B-Cell Lymphomas; B-Cell Neoplasm; B-Cell NonHodgkins Lymphoma; B-Lymphocytes; Benign; Binding; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Bone Marrow; Bone marrow biopsy; Cell surface; Cells; Chromogenic in situ Hybridization; Chronic; Clinical; Clonality; Color; Computer software; Cytogenetic Analysis; Detection; Development; Diagnosis; Diagnostic; Differential Diagnosis; Disease; Evaluable Disease; Evaluation; Extranodal; Flow Cytometry; Follicular Lymphoma; Formalin; Fresh Tissue; Goals; Health; Hyperplasia; Image Analysis; Immunohistochemistry; In Situ Hybridization; Inflammation; Laboratories; Light; Light-Chain Immunoglobulins; Lymphoid; Lymphoma; Lymphoproliferative Disorders; Manuals; Measures; Messenger RNA; Methods; Molecular; Molecular Cytogenetics; Molecular Diagnostic Testing; Morphology; Non-Hodgkin' s Lymphoma; Paraffin Embedding; Pathologic; Pathologist; Pathology; Performance; Phase; Probability; Procedures; Proteins; Protocols documentation; RNA; Reagent; Savings; Series; Slide; Small Business Innovation Research Grant; Small-Cell Lymphoma; Specimen; Staining method; Stains; Surface; Surface Immunoglobulins; Technology; Time; Tissues; Translational Research; Validation; Variant; base; cohort; cost; digital; instrument; kappa-Chain Immunoglobulins; large cell Diffuse non-Hodgkin' s lymphoma; lymphoid hyperplasia; mucosa-associated lymphoid tissue lymphoma; novel; novel diagnostics; personalized medicine; prospective; single molecule; success; tool

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.

描述（由申请人提供）：免疫球蛋白（IG）轻链限制（LCR）检测是鉴别诊断中的重要分子诊断工具，其中包括淋巴样增生，非典型淋巴性增生，慢性炎症，慢性炎症和B细胞Neoplasms。可以通过流式细胞仪（FCM）检测到LCR作为细胞表面上的异常kappa：lambda比。但是，FCM需要新鲜的（未固定）组织。一些B细胞淋巴瘤缺乏FCM可检测到的表面Ig表达，有些表现出非特异性抗体结合。通常，只有福尔马林固定的石蜡包埋（FFPE）组织可用于病理评估。 FFPE组织中LCR鉴定的当前方法包括免疫组织化学（IHC）和常规的发色原位杂交（CISH），分别测量了Kappa或Lambda蛋白和mRNA。但是，IHC和常规CISH缺乏足够的敏感性来检测许多非霍奇金淋巴瘤（NHL），包括最常见的NHL变体 - 卵泡淋巴瘤，弥漫性大B细胞淋巴瘤，小B淋巴细胞淋巴瘤淋巴瘤，小型B淋巴细胞淋巴瘤和麦芽瘤外膜瘤。 RNASCOPE是一种新型的RNA原位杂交技术，具有单分子检测敏感性，并使用成色或荧光检测与FFPE组织完全兼容。我们已经为LCR开发了手动明亮场Kappa和Lambda双链RNA ISH ISH测定法，该测定法证明了对所有NHL亚型的广泛适用性。在这个项目中，我们将开发一种具有数字病理学解释的完全自动化的双工RNASCOPE LCR测定法，这应该极大地促进这种新颖测定法的临床采用。

项目成果

Ultrasensitive automated RNA in situ hybridization for kappa and lambda light chain mRNA detects B-cell clonality in tissue biopsies with performance comparable or superior to flow cytometry.

• DOI: 10.1038/modpathol.2017.142
• 发表时间: 2018-03
• 期刊: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc
• 作者: Guo L;Wang Z;Anderson CM;Doolittle E;Kernag S;Cotta CV;Ondrejka SL;Ma XJ;Cook JR
• 通讯作者: Cook JR