BIOMED-2系统IgH引物的筛选验证及相关基因重排研究

发布时间：2018-03-04 09:06

本文选题：非霍奇金淋巴瘤　切入点：免疫球蛋白重链　出处：《第一军医大学》2006年硕士论文　论文类型：学位论文

【摘要】：一、研究背景和目的淋巴瘤病理诊断是临床病理诊断的难题之一，2001年世界卫生组织(WHO)定义恶性淋巴瘤中每一个独立疾病(或类型)需要结合形态学、免疫表型、遗传学和临床特点来确定，特别强调基因重排检测技术辅助诊断淋巴瘤的重要性。大部分可疑的高度反应增生性淋巴组织，可以通过组织形态学和细胞形态学结合免疫组化或流式细胞技术检测免疫表型来与恶性淋巴瘤甄别。但还有大约5～15％的病例难以确诊。原则上，恶性淋巴瘤的瘤细胞都来自一个共同的克隆，所以能够依靠克隆性鉴定来进行辅助诊断。淋巴细胞恶性肿瘤以B细胞来源为主，约占90％，免疫球蛋白重链基因(IgH)分析方法主要有DNA印迹法(Southern blot)、PCR、测序等。以PCR技术为基础的方法以其简便、高效、廉价及适用面广而在淋巴瘤基因重排研究方面越来越受重视。但不同研究文献中所采用的引物、PCR条件、PCR产物检测方法等都不尽相同，结果也有差异。难以找到适于常规临床病理诊断的标准程序，影响了PCR技术的应用价值。 2003年欧洲七国47个研究所的病理学家、分子生物学家及其它研究人员经过四年半的研究发表了标准化引物系统(BIOMED-2)，建立了用于克隆性分析的基因重排及染色体易位检测的PCR方法学和引物设计的标准化，可检测出几乎所有克隆性T、B细胞克隆细胞群乃至高度体细胞突变B细胞淋巴瘤／白血病，实现了前所未有高检出率。BIOMED-2引物系统是划时代的，其设计思想十分先进，，将该技术进一步完善并及早运用于临床已势在必行，但该系统过于庞大复杂，目前在国内难以推广应用，更不利于临床诊断的使用。本研究以淋巴瘤
[Abstract]:I. background and purpose of the study. The pathological diagnosis of lymphoma is one of the difficult problems in clinicopathological diagnosis. In 2001, the World Health Organization (WHO) defined each individual disease (or type) in malignant lymphoma to be determined by combining morphology, immunophenotype, genetics and clinical characteristics. Emphasizing in particular the importance of gene rearrangement techniques to assist in the diagnosis of lymphoma. Most suspicious highly reactive proliferative lymphoid tissues, Histomorphology and cell morphology combined with immunohistochemistry or flow cytometry can be used to detect the immunophenotype of malignant lymphoma. But there are still about 55% of the cases that are difficult to diagnose. In principle, Malignant lymphoma cells are derived from a common clone, so can rely on clonal identification to assist the diagnosis. The immunoglobulin heavy chain gene (Ig) analysis method mainly includes DNA blotDNA, sequencing and so on. The method based on PCR technique is simple and efficient. More and more attention has been paid to the gene rearrangement of lymphoma due to its low cost and wide application. It is difficult to find a standard procedure suitable for routine clinicopathologic diagnosis, which affects the application value of PCR technique. In 2003, pathologists at 47 institutes in seven European countries, After four and a half years of research, molecular biologists and other researchers have published a standardized primer system, BIOMED-2, and established PCR methodology and standardization of primer design for gene rearrangement and chromosome translocation detection for clonal analysis. It can detect almost all clone T B cell colony and even high somatic mutation B cell lymphoma / leukemia. It is epochal to realize the unprecedented high detection rate of BIOMED-2 primer system, and its design idea is very advanced. It is imperative to improve the technique and apply it to clinical practice as soon as possible, but the system is too large and complex, so it is difficult to be popularized and applied in China at present, which is even more unfavorable for clinical diagnosis.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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