遗传病推断复合扩增体系的构建  

A Novel Multiplex PCR Assay for the Inference of Genetic Diseases

作　　者：孙启凡[1,2,3] 赵蕾[1,2,3] 徐颖[1,4] 王玮[1,5] 龙源[1] 马纪强[1] 魏以梁[6,7] 赵兴春[1,2,3] 叶健[1,2,3] 李彩霞[1,2,3] 

机构地区：[1]公安部物证鉴定中心,北京100038 [2]北京市现场物证检验工程技术研究中心,北京100038 [3]法医遗传学公安部重点实验室,北京100038 [4]中国刑警学院,沈阳110854 [5]山西医科大学,太原030024 [6]清华大学医学院生物医学工程系,北京100084 [7]天津医科大学表观遗传中心,天津300070

出　　处：《刑事技术》2015年第5期364-367,共4页Forensic Science and Technology

基　　金：公安部科技强警基础工作专项项目(No.2013GABJC037)

摘　　要：目的通过对人类DNA进行突变基因检测推断该DNA供者是否患特定遗传病,建立对DNA样本突变检测的方法,运用于法庭科学的DNA来源人特征刻画领域,为案件侦查提供线索。方法根据遗传性耳聋、β-地中海贫血、葡萄糖-6-磷酸脱氢酶缺乏症三类遗传疾病各突变位点在中国人群中的出现概率,挑选了20个位点,设计引物,利用等位基因特异性PCR和毛细管电泳技术构建推断这三类遗传疾病的PCR检测体系。结果构建了包含20个突变位点,能够同时检测遗传性耳聋、地中海贫血、葡萄糖-6-磷酸脱氢酶(G6PD)三类遗传疾病的毛细管电泳复合扩增实验体系。结论在法医学实验室开展检材DNA来源人的疾病特征推断是可行的。通过进一步研究发展,该技术有望在刑事案件侦查中发挥作用。Objective Genetic disease, usually controlled by pathogenic genes, occurs over mutation in the relevant genetic material. Theoretically, scientists can infer from the DNA typing whether a subject is prone to some kind of genetic diseases because of the regionality and/or inheritance of most these illnesses able to link the DNA donor with certain zone of the typed DNA. Therefore, the inference of unknown supplier of evidential substance, based on the information of some genetic diseases parsed with the relative DNA, may become an important research focus in forensic community due to the common fact that target suspects and/or any other clues are not easily exposed through the biologic samples collected from the scenes in criminal cases. Here, a simple and efficient method was tried to establish for the detection of certain pathogenic genes. Methods Allele specific PCR （ASPCR） was conducted in two aliquots of each separately subjected to normal and mutant amplification with the allele specific primers designed according to the sequenced gene, and followed by capillary electrophoresis for gene sequencing to get its genotype. Results A newly developed panel of 20 genes, synchronously detecting three kinds of genetic diseases based on allele specific PCR technique and capillary electrophoresis, were presented. The panel contained l 1 disease genes of Genetic Deafness （one of the most common sensory disorders that affect communication）, 6 genetic loci of Beta Mediterranean anemia （of high incidence in China＇s southern parts such as Guangdong, Guangxi and Hong Kong） and 3 loci of Glucose-6-phosphate dehydrogenase （G6PD） deficiency （having its higher incidence in southern China, too）. Conclusions The multiplex assay, performed with PCR and the capillary electrophoresis available at most forensic genetic laboratories, is very likely to be a convenient and cost-effective choice for criminal investigations.

关 键 词：法医遗传学 等位基因特异性PCR 毛细管电泳 遗传性耳聋 Β-地中海贫血 葡萄糖-6-磷酸脱氢酶缺乏症 

分 类 号：DF795.2[医药卫生—法医学]