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New method for nucleic acid amplification on pathologic tissue section of malignant lymphoma using LAMP method

LAMP法恶性淋巴瘤病理组织切片核酸扩增新方法

基本信息

批准号：
18590341
负责人：
NAKAMINE Hirokazu
金额：
$ 1.67万
依托单位：
Nara Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590341/
关键词：
Malignant lymphoma LAMP method in situ nucleic acid amplification Follicular lymphoma bcl-2 rearrangement FISH method in situ 核酸増幅 bc1-2再構成

项目摘要

Classification of malignant lymphoma is important for selection of treatment modality as well as for investigation of pathogenesis. Because B-cell lymphomas consist of several types which are correspond well to chromosomal/genetic abnormalities, more accurate classification can be performed when these abnormalities are detectable at daily pathology practice. However ; it is rather general in Japan that the first biopsy for pathological diagnosis is often performed at the regional hospitals rather than at the specialized medical institutions. The patients are transferred to the latter institutions where the second biopsy is sometimes required for examination of chromosomal/genetic abnormalities. This is due to the limitation and difficulty in performing such examination on paraffin-embedded tissue. To overcome this problem, we attempted to establish a new method for microscopic visualization of these abnormalities by amplification of genes involved in them on microscopic glass slides. We employed loop-mediated isothermal amplification (LAMP) instead of conventional PCR for this in situ DNA amplification, because LAMP method has several advantages over the conventional PCR for this purpose, including lower degree of tissue damages by using lower temperature, use of small-sized DNA polymerase, and larger size of amplification products. We fried to in situ amplify the rearranged bcl-2/IgH genes as a result of t (14 ; 18) which are present in more than 80% cases of follicular lymphoma. However, this attempt appears to be more difficult than initially thought and satisfactory amplification has not been obtained so far In the literature, only a single report of successful detection of point mutation of some gene by this in situ LAMP (Ikeda, et. al. Pathol Int 57, 594-599, 2007), indicating that there may be unexpected problems. Through this trial, however ; several minor findings regarding B-cell lymphoma could be obtained.

恶性淋巴瘤的分型对治疗方式的选择和发病机制的研究具有重要意义。由于B细胞淋巴瘤由几种类型组成，这些类型与染色体/遗传异常很好地对应，当这些异常在日常病理学实践中可检测时，可以进行更准确的分类。然;在日本，用于病理诊断的第一次活组织检查通常在地区医院而不是在专门的医疗机构进行，这是相当普遍的。病人被转移到后一种机构，在那里有时需要进行第二次活组织检查，以检查染色体/遗传异常。这是由于在石蜡包埋组织上进行这种检查的局限性和困难。为了克服这个问题，我们试图建立一种新的方法，通过在显微镜载玻片上扩增与这些异常有关的基因来进行显微镜可视化。我们采用环介导等温扩增（LAMP）代替常规PCR进行原位DNA扩增，因为LAMP方法与常规PCR相比具有几个优点，包括使用较低的温度，使用小尺寸的DNA聚合酶，以及扩增产物的较大尺寸。本研究采用原位扩增技术检测了80%以上滤泡性淋巴瘤中存在的t（14 ; 18）引起的bcl-2/IgH基因重排。然而，这种尝试似乎比最初想象的更困难，并且迄今为止还没有获得令人满意的扩增。在文献中，只有一个通过这种原位LAMP成功检测某些基因的点突变的报道（Ikeda，et.等人，Pathol Int 57，594-599，2007），表明可能存在意想不到的问题。然而，通过这项试验;可以获得有关B细胞淋巴瘤的一些小发现。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

B細胞リンパ腫病理診断の基本.びまん性大細胞型B細胞リンパ腫を中心に

B细胞淋巴瘤病理诊断基础.聚焦弥漫性大B细胞淋巴瘤

DOI：
发表时间：
2007
期刊：
臨床血液 48
影响因子：
0
作者：
Zhang;B;Morii;E;et. al.;Oka K;中峯寛和
通讯作者：
中峯寛和

Prognostic impact of chromosomal alteration of 3q27 on nodal B-cell lymphoma: Correlation with histology, immunophenotype, karyotype, and clinical outcome in 329 consecutive patients

DOI：
10.1016/j.leukres.2006.11.004
发表时间：
2007-09-01
期刊：
LEUKEMIA RESEARCH
影响因子：
2.7
作者：
Niitsu, Nozomi;Okamoto, Masataka;Miura, Ikuo
通讯作者：
Miura, Ikuo

Follicular lymphoma frequently originates in the salivary gland.

滤泡性淋巴瘤通常起源于唾液腺。

DOI：
发表时间：
2006
期刊：
Pathol Int. 56・10
影响因子：
0
作者：
Nakamura S;Ichimura K;Sato Y;Nakamura S;Nakamine H;Inagaki H;Sadahira Y;Ohshima K;Sakugawa S;Kondo E;Yanai H;Ohara N;Yoshino T
通讯作者：
Yoshino T

末梢性T細胞リンパ腫再発時にT/B両クローンが検出されEBV陽性細胞の増殖を認めた1例

外周T细胞淋巴瘤复发时同时检测到T、B克隆并观察到EBV阳性细胞增殖一例