The Application of DNA Polymorphism to Paternity Testing

DNA多态性在亲子鉴定中的应用

• 批准号: 04670362
• 负责人: SHINOZUKA Tatsuo
• 金额: $ 1.09万
• 依托单位: KEIO 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670362/
• 关键词: DNA polymorphism; Paternity; Restriction Fragment Length Polymorphisms (RFLPs); Polymerase Chain Reaction (RCR); HLA DQ alpha Typing; RFLPs法; HLADQalpha型; RFLP法; HLADQα

DNA was extracted from cryopreserved lymphocytes after paternity testing by the phenol-chloroform method or commercial DNA extraction kits. Paternity analysis has been performed by a simple method for nonradioactive DNA fingerprinting. The method utilizes a minisatelite probe MZ1.3 labeled with digoxigenin, which are detected using an anti-digoxigenin-alkaline phosphatase complex. This system detected DNA restriction fragments in more than 10mug of DNA from lymphocytes.On the other hand, the experiment has been performed to evaluate typing of the HLA DQ a gene by polymerase chain reaction (PCR) amplification of the gene and subsequent hybridization with sequence-specific oligonucleotide probe (Ampli Type kit). The sensitivity to detect DNA extraction from the lymphocytes was evaluated by amplifying various amounts of purified DNA.HLA DQ a typing were detected from 1-20ng as amounts of template DNA from paternity test.

亲子鉴定后，用酚-氯仿法或市售DNA提取试剂盒从冻存的淋巴细胞中提取DNA。亲子鉴定是通过一种简单的非放射性DNA指纹法进行的。该方法使用地高辛标记的小卫星探针MZ1.3，使用抗地高辛-碱性磷酸酶复合物进行检测。该系统可检测10 μ g以上淋巴细胞DNA中的DNA限制性片段，另一方面，本实验通过聚合酶链反应（PCR）扩增HLA DQ α基因，然后与序列特异性寡核苷酸探针（Ampli Type试剂盒）杂交，对HLA DQ α基因进行了分型评价。以1- 20 ng的DNA作为亲子鉴定的模板DNA，检测HLA DQ a型。

项目成果

篠塚達雄、他: "HLA検査後の凍結保存リンパ球からのDNA抽出とHLADQα型の検討" 法医学の実際と研究. 36. 17-20 (1993)

Tatsuo Shinozuka 等人：“HLA 检测和 HLADQα 型检查后从冷冻保存的淋巴细胞中提取 DNA”《法医实践与研究》36. 17-20 (1993)。

篠塚達雄、他: "HLA検査後の凍結保存リンパ球からのDNA抽出とHLADQalpha型の検討" 法医学の実際と研究. 36. 17-20 (1993)

Tatsuo Shinozuka 等人：“HLA 测试和 HLADQalpha 型检查后从冷冻保存的淋巴细胞中提取 DNA”《法医实践与研究》36. 17-20 (1993)。

原 正昭: "HLA検査後のリンパ球からのDNA抽出法とDNA解析への適用" DNA多型. (1993)

Masaaki Hara：“HLA测试后从淋巴细胞中提取DNA的方法及其在DNA分析中的应用”DNA多态性（1993）。

Hara, M., Yabe, K., Saito, K., Murai, T., Watanabe, H., Shinozuka, T.and Yanagida, J.: "The application of DNA polymorphisms to paternity testing (1). DNA extraction from lymphocytes and application to paternity testing." DNA POLYMORPHISM. 1. 41-48 (1993)

Hara, M.、Yabe, K.、Saito, K.、Murai, T.、Watanabe, H.、Shinozuka, T. 和 Yanagida, J.：“DNA 多态性在亲子鉴定中的应用 (1)。DNA 提取

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

Shinozuka, T.、Hara, M.、Takei, S.、Watanabe, H.、Kuroda, N.、Kurihara, K. 和 Yanagida, J.：“亲子鉴定淋巴细胞 DNA 提取和 HLA DQ α 基因座检测