Microsatellite instability in mononuclear cells from non-tumorigenetic human tonsils and its forensic applications

非致瘤性人扁桃体单核细胞的微卫星不稳定性及其法医学应用

• 批准号: 10670402
• 负责人: KOBAYASHI Ryo
• 金额: $ 2.11万
• 依托单位: Tokyo 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670402/
• 关键词: microsatellite; PCR; personal identification; DNA; TH01; tonsil; EBV

Microsatellites consisted of tandem repeated sequence motifs of 1 to 6 nucleotides in length are highly polymorphism and scattered more or less randomly throughout the genome. The use of polymorphism microsatellite DNA sequences has become extremely important in genetic mapping studies and in the diagnosis of human genetic diseases. Microsatellite instability (MSI) has been reported to be a landmark of some tumors of the colon and rectum and is characterized by small deletions or expansions within short tandem repeats in tumor DNA as compared with matching normal DNA.In this project, we have tried to find the presence of MSI in healthy individuals and to examine the application of forensic cases. TH01 locus was selected as a microsatellite and determined at the mononuclear cells isolated from tonsils. Seminested PCR was performed to determine TH01 types using single cells. One hundred and seventy-eight out of 450 cells could be typed (39.6%) and the type of only 1 cell (0.56%) was mutated. This result indicates that a microsatellite alteration at TH01 locus may be observed in the cells proliferated vigorously in tissues. However, there is little probability that microsatellite typing will be incorrect using a DNA template that includes mutated DNA at 0.56% of cells.

微卫星是由长度为1 ~ 6个核苷酸的串联重复序列组成，具有高度多态性，在基因组中随机分布。多态性微卫星DNA序列的使用在遗传作图研究和人类遗传疾病的诊断中已变得极其重要。微卫星不稳定性（Microsatellite instability，MSI）是结直肠肿瘤的一个标志性特征，其特征是肿瘤DNA中短串联重复序列与正常DNA相比出现小的缺失或扩增。选择TH 01位点作为微卫星，并在从扁桃体分离的单个核细胞上进行测定。使用单细胞进行半巢式PCR以确定TH 01类型。450个细胞中有178个可分型（39.6%），只有1个细胞（0.56%）的类型发生突变。这一结果表明，在组织中增殖旺盛的细胞中可能观察到TH 01位点的微卫星改变。然而，使用包含0.56%细胞突变DNA的DNA模板，微卫星分型不正确的可能性很小。

项目成果

佐藤耕一: "Primer extension preamplification(PEP)法を用いたDNA型判定について -型判定の正確さと検出限界の検討-" 日法医誌. 52. 184-190 (1998)

Koichi Sato：“关于使用引物延伸预扩增 (PEP) 方法进行 DNA 分型 - 分型准确性和检测限的检查 -”《日本法医学杂志》52. 184-190 (1998)。

Kobayashi,R.: "Detection of Epstein-Barr virus infection in the epithelial cells and lymphocytes of non-neoplastic tonsils by in situ hybridization and in situ PCR" Arch Virol. 143. 803-813 (1998)

Kobayashi,R.：“通过原位杂交和原位 PCR 检测非肿瘤性扁桃体上皮细胞和淋巴细胞中的 Epstein-Barr 病毒感染”Arch Virol。

Itoh,Y.: "Discrepant results of ABO blood grouping obtained by serologic and PCR techniques from highly putrefied semen"Prog Foren Genet. 8. 548-551 (2000)

Itoh,Y.：“通过血清学和 PCR 技术从高度腐烂的精液中获得的 ABO 血型结果不一致”Prog Foren Genet。

池田哲也: "Epstein-Barrウイルスと突発性間質性肺炎との関連について"東医大誌. 58. 527-532 (2000)

池田哲也：“EB 病毒与突发性间质性肺炎的关系”，东京医科大学学报 58. 527-532 (2000)。

Itoh Y: "DNA typing of ABO and Lewis blood groups using whole genome amplification"DNA Polymorphism. 7. 237-240 (1999)

Itoh Y：“使用全基因组扩增对 ABO 和 Lewis 血型进行 DNA 分型”DNA 多态性。