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The study of virus DNA polymorphisms and its application to paternity testing

病毒DNA多态性研究及其在亲子鉴定中的应用

基本信息

批准号：
05670400
负责人：
KOBAYASHI Ryo
金额：
$ 1.34万
依托单位：
Juntento University School of Medicine
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 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670400/
关键词：
personal identification paternity testing Epstein-Barr virus short tandem repeat DNA PCR polymorphism polymorphism 扁桃腺 in situ hybridization

项目摘要

In this project, we have tried to find the DNA markers that might be acquired in persons after their birth and be named acquired DNA markers. We have also tried to apply them for personal identification and paternity testing. In 1993, the third complementarity-determining region (CDR3) of immunoglobulin heavy chain gene was selected as one of acquired DNA markers, because this gene was rearranged during B-cell differentiation. However, the life span of the mature B-cells whose immunoglobulin gene was rearranged is generally considered to be about 2 weeks. Therefore, we did not think that this gene was the acquired DNA marker that could identify individuals. In 1994, we selected virus genes, especially Epstein-Barr (EB) virus as the acquired DNA marker, because this virus is a human herpesvirus that is carried by more than 90% of human population worldwide. We could detect the EB virus DNA in 92% of 98 DNA samples from peripheral blood lymphocytes using PCR that amplified BamW region of EB virus. EB virus DNA might be the acquired DNA marker, because this virus infects persons after their birth, and could be easily differentiable from the host DNA.In 1995, we focused the polymorphism of EB virus. EB virus has several short tandem repeat sequences and it is known that part of these sequences sometimes mutate during asymptomatic infection. So, we amplified the region of short tandem repeats of EB virus in the DNA isolated from peripheral blood lymphocytes using PCR and could observe the polymorphisms of EB virus in individuals. We concluded that EB virus might be considered as one of acquired DNA markers. However, we don't know how many strains of EB virus there are in persons, or how long EB virus is infected. These subjects will be had to elucidated.

本课题试图寻找人在出生后可能获得的DNA标记，并命名为获得性DNA标记。我们还试图将其用于个人身份识别和亲子鉴定。1993年，免疫球蛋白重链基因的第三互补决定区（CDR3）被选为获得性DNA标记之一，因为该基因在B细胞分化过程中发生重排。然而，其免疫球蛋白基因重排的成熟B细胞的寿命通常被认为是约2周。因此，我们不认为该基因是可以识别个体的后天DNA标记。1994年，我们选择了病毒基因，特别是EB病毒作为获得性DNA标记，因为这种病毒是一种人类疱疹病毒，在全球90%以上的人群中携带。用扩增EB病毒BamW区的PCR方法对98例外周血淋巴细胞DNA进行检测，检出率为92%。EB病毒DNA可能是获得性DNA标记，因为这种病毒在出生后感染人，并且很容易与宿主DNA区分。EB病毒具有几个短串联重复序列，并且已知这些序列的一部分有时在无症状感染期间发生突变。因此，本研究采用PCR技术扩增了EB病毒短串联重复序列的区域，并观察了EB病毒在个体中的多态性。因此，EB病毒可作为获得性DNA标志物之一。然而，我们不知道人体内有多少株EB病毒，也不知道EB病毒感染了多长时间。这些问题必须加以阐明。