Development of mutation analysis system by multiplex PCR and suspension array for paraffin-embedded autopsy samples
石蜡包埋尸检样本多重PCR和悬浮阵列突变分析系统的开发
基本信息
- 批准号:21590411
- 负责人:
- 金额:$ 3.08万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2009
- 资助国家:日本
- 起止时间:2009 至 2011
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Objective : To establish a suspension array-based analysis system for determining mitochondrial haplogroups, using DNA purified from formalin-fixed paraffin-embedded(FFPE) autopsy tissues. Background : Because the evolutionary rate of mitochondrial DNA(mtDNA) is much higher than that of nuclear DNA, and because mtDNA is highly polymorphic, mitochondrial single nucleotide polymorphisms(mtSNPs) are expected to contribute more extensively to the functional differences among individuals than those SNPs in the nuclear DNA. Design/Methods : We carefully purified total DNA from FFPE autopsy tissues by using ethanol precipitation. By using our system, we could estimate 7 major mitochondrial haplogroups for Japanese containing 17 subhaplogroups belonging to 2 super haplogroups(M and N), based on information obtained from the comprehensive analysis of mtSNPs in the coding region of mtDNA. We first tested approximately 100 DNA samples from blood and FFPE tissues of autopsy subjects whose haplogroups had been identified by DNA analysis of frozen-kidney tissues. To examine more fully the utility of the system, we analyzed 5, 161 DNA samples from FFPE tissues. Results : We could analyze 4, 485(87%) of them successfully. However, we could not analyze the remainder(13%) because of an insufficient PCR amplification due to severe DNA fragmentation. We had previously revealed the haplogroup frequencies of 1, 500 frozen-kidney DNA samples. In comparison, the haplogroup D4a was overestimated, but the haplogroups M7a, D4b1b and N9b were underestimated, by the present system we developed. Conclusions : We have established a method to purify DNA from FFPE tissues and developed a suspension array-based analytic system to estimate major haplogroups of Japanese individuals. We believe that our system for the classification of mitochondrial haplogroups will be very helpful not only for pathogenetical studies, but also for epidemiological cohort studies.
目的:利用从福尔马林固定石蜡包埋(FFPE)尸检组织中提取的DNA,建立基于悬浮阵列的线粒体单倍群分析系统。背景:由于线粒体DNA(MtDNA)的进化速度远高于核DNA,且线粒体DNA具有高度的多态性,线粒体单核苷酸多态(MtSNPs)可能比核DNA中的SNPs对个体间的功能差异有更广泛的贡献。设计/方法:采用乙醇沉淀法从FFPE尸检组织中提取总DNA。根据对mtDNA编码区mtSNPs的综合分析,我们可以估计出7个主要的日本人线粒体单倍群,包括属于2个超单倍群(M和N)的17个亚单群。我们首先从尸检受试者的血液和FFPE组织中检测了大约100个DNA样本,这些受试者的单倍型已经通过冰冻肾脏组织的DNA分析确定。为了更全面地检验该系统的实用性,我们分析了来自FFPE组织的5161个DNA样本。结果:成功分析4485例(87%)。然而,我们无法分析其余的(13%),因为由于严重的DNA片段化而导致PCR扩增不充分。我们之前公布了1,500个冷冻肾脏DNA样本的单倍组频率。相比之下,我们开发的系统高估了单倍群D4a,而低估了单倍群M7a、D4b1b和N9b。结论:我们建立了一种从FFPE组织中纯化DNA的方法,并开发了一种基于悬浮阵列的分析系统来估计日本人的主要单倍群。我们相信,我们的线粒体单倍群分类系统不仅对病因学研究非常有帮助,而且对流行病学队列研究也很有帮助。
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)
ピルビン酸療法
丙酮酸盐疗法
- DOI:
- 发表时间:2010
- 期刊:
- 影响因子:0
- 作者:Masayo Minami;Tomomi Kato;Yoshiki Miyata;Toshio Nakamura and Quan Hua;田中雅嗣
- 通讯作者:田中雅嗣
Association of a Polymorphism of BTN2A1 With Hypertension in Japanese Individuals
- DOI:10.1038/ajh.2011.74
- 发表时间:2011-08-01
- 期刊:
- 影响因子:3.2
- 作者:Horibe, Hideki;Kato, Kimihiko;Yamada, Yoshiji
- 通讯作者:Yamada, Yoshiji
The establishment of an expanded simultaneous swift diagnosing system for mitochondrial DNA pathological mutations.
建立了线粒体DNA病理突变的扩展同步快速诊断系统。
- DOI:
- 发表时间:2009
- 期刊:
- 影响因子:0
- 作者:Xie ZH;et. al.;Nishigaki Y
- 通讯作者:Nishigaki Y
Polymorphisms in the Control Region of Mitochondrial DNA Associate with Elite Japanese Athlete Status
线粒体 DNA 控制区的多态性与日本优秀运动员的地位相关
- DOI:
- 发表时间:2011
- 期刊:
- 影响因子:0
- 作者:Eri Mikami;Noriyuki Fuku;Hideyuki Takahashi;Nao Ohiwa;Yannis P Pitsiladis;Mitsuru Higuchi;Takashi Kawahara;Masashi Tanaka
- 通讯作者:Masashi Tanaka
Comprehensive Mitochondrial Haplogroup Analysis System Based On Suspension-Array Technology Using DNA From Formalin-Fixed Paraffin-Embedded Autopsy Tissues
基于悬浮阵列技术的综合线粒体单倍群分析系统,使用福尔马林固定石蜡包埋尸检组织中的 DNA
- DOI:
- 发表时间:2012
- 期刊:
- 影响因子:0
- 作者:Yutaka Nishigaki;Hiroko Hosoya;Makiko Naka-Mieno;Tomio Arai;Motoji Sawabe;Masashi Tanaka
- 通讯作者:Masashi Tanaka
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HOSOYA Hiroko其他文献
HOSOYA Hiroko的其他文献
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{{ truncateString('HOSOYA Hiroko', 18)}}的其他基金
Neural recognition molecules, NrCAM and Contactin, during development and aging.
发育和衰老过程中的神经识别分子 NrCAM 和 Contactin。
- 批准号:
10832010 - 财政年份:1998
- 资助金额:
$ 3.08万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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Grant-in-Aid for Encouragement of Young Scientists (A)