The Application of DNA Plymorphism to Paternity Testing-DNA Extraction from the Small Volume of Lympocytes Left over after HLA Typing-

DNA多态性在亲子鉴定中的应用-HLA分型后残留的少量淋巴细胞中提取DNA-

• 批准号: 05670398
• 负责人: HARA Masaaki
• 金额: $ 1.41万
• 依托单位: Saitama Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670398/
• 关键词: formain-fixed; lymphocytes; DNA extraction; HLA Typing; HLA-DQalpha typing; Paternity Test; HLA; HLA DQα; ハプロタイプ; PM-PCR

Our study in 1993 focused on evaluation of the method of DNA extraction from lymphocytes fixed in formalin. However, we found that DNA extracted from formalin-fixed lymphocytes rapidly undergoes degradation to lower molecular from owing to the effect of the formalin.Therefor, in 1994 we extracted DNA from the smoll residual portion of lymphocytes used for HLA typing, and perfomed HLADQalpha typing on the DNA obtained.We performed HLA typing in 26 lineages and identified the A-C-B haplotypes. After that, the residual small volume of lymphocytes was used enough to extract DNA,and the extracted DNA was analyzed for HLADQalpha. All specimens were large to be typed.We then investigated the linkage between the A-C-B haplotypes and the DQalpha type and identified a strong linkage between A24-CwX-Bw52 and the DQalpha1.3 typ, and A24-Cw7-B7 and DQalpha1.1.These results show that specimens needed for DNA analysis can be obtained from the Small Volume of Lympocytes Left over after HLA Typing.

我们在1993年的研究重点是评价从福尔马林固定的淋巴细胞中提取DNA的方法。然而，我们发现从福尔马林固定的淋巴细胞中提取的DNA在福尔马林的作用下会迅速降解为低分子。为此，我们于1994年从用于HLA分型的淋巴细胞的Smoll残留部分中提取DNA，并对获得的DNA进行HLADQAlpha分型。我们对26个谱系进行了HLADQpha分型，并鉴定了A-C-B单倍型。然后，利用剩余的小体积淋巴细胞提取足够的DNA，并对提取的DNA进行HLADQpha分析。结果表明，A24-CwX-BW52与DQalpha1.3、A24-Cw7-B7与DQalpha1.1之间存在较强的连锁关系。这些结果表明，DNA分析所需的标本可以从HLA分型后剩余的少量淋巴细胞中获得。

项目成果

Hara, M., Takada, A., Yabe, K., Murai, T., Watanabe, H., Shinozuka, T., Yanagida, J., Yamamoto, Y., and Tajima, T.: "LDLR, GYPA, HBGG, D7S8, GC, Genotyping by PM-PCR and Its application to paternity testing." DNA POLYMORPHISM. 3. 181-186 (1995)

Hara, M.、Takada, A.、Yabe, K.、Murai, T.、Watanabe, H.、Shinozuka, T.、Yanagida, J.、Yamamoto, Y. 和 Tajima, T.：“LDLR，GYPA

Shinozuka, T.Takei, S., Kurihara, K., Yanagida, J., Hara, M.and Watanabe, H.18GB03 : The application of DNA polymorphisms to paternity testing (3).: DNA POLYMORPHISM. 2. 99-102 (1994)

Shinozuka，T.Takei，S.，Kurihara，K.，Yanagida，J.，Hara，M.和Watanabe，H.18GB03：DNA多态性在亲子鉴定中的应用(3)。：DNA多态性。

原正昭: "HLA-DQα部位のDNA型とHLA-A-C-Bハプロタイプとの連鎖について" 日本法医学雑誌. 49(1). 59-59 (1995)

Masaaki Hara：“关于 HLA-DQα 位点的 DNA 类型与 HLA-A-C-B 单倍型之间的联系”《日本法医学杂志》49(1) (1995)。

Shinozuka, T., Hara, M., Takai, S., Watanabe, H., Kuroda, N., Kurihara, K.and Yanagida, J.: "DNA extraction from lymphocytes of paternity testing and detection of HLADQalpha locus by PCR applifcation." Res. Pract. Forens. Med.36. 17-20 (1993)

Shinozuka, T.、Hara, M.、Takai, S.、Watanabe, H.、Kuroda, N.、Kurihara, K. 和 Yanagida, J.：“从亲子鉴定淋巴细胞中提取 DNA 并通过 PCR 检测 HLADQalpha 基因座

原正昭: "MP-PCR法によるLDKR,GYPA,HBGG,D7S8,GCの遺伝子型判定とその親子鑑定への応用" DNA多型. 3(発表予定). (1995)

Masaaki Hara：“通过 MP-PCR 方法进行 LDKR、GYPA、HBGG、D7S8、GC 的基因分型及其在亲子鉴定中的应用”DNA 多态性 3（待出版）。