ABO基因CASPA分型方法的研究

发布时间：2018-08-03 21:54

【摘要】：前言 ABO血型是第一个被发现的人类血液遗传标记系统。1990年,Yamamoto等人首先报道了ABO基因的核酸序列,并且确定了决定ABO血型表型的各等位基因的碱基序列差异点,这一发现是在DNA水平上检测ABO基因型的遗传学基础。目前应用的ABO基因分型技术大概可以分为扩增限制性片段长度多态性法(PCR-RFLP)、单链构象多态性法(SSCP)、PCR直接测序法、等位基因特异性扩增法(ASA)、等位基因特异性引物消耗法(CASPA)等几类技术方法。其中,等位基因特异性扩增法是当前在法医物证检验鉴定领域研究和应用最为广泛的一种ABO基因分型方法,该方法具有灵敏、简便、快捷等特点。本项研究所选择的等位基因特异性引物消耗法(CASPA法)就是近期在ASA法基础上研究建立的一种新的ABO基因分型方法。该方法的优点是:1.由于采用毛细管电泳仪检测扩增后剩余的荧光标记引物,所以方法更加简单、快速、灵敏;2.由于其等位基因特异性引物长度与其扩增产物相比较要短的多,所以电泳分离时间缩短;3.由于所检测的是扩增后剩余引物,所以该方法适合在降解生物检材的ABO基因分型中应用。本项研究在Watanabe等人报道的等位基因特异性引物消耗法的基础上,在引物设计上进行了改进,并进行了法医常见生物物证的适用性研究,现将研究结果报告如下: 实验材料分别被荧光染料6-FAM和HEX标记的6条长度不等的等位基因特异性引物以及1条无标记物的共用引物。已知ABO基因型的DNA提取液。146份中国汉族无关个体血液斑样品。混合斑、烟蒂、汗液斑、肌肉、软骨、股骨、带毛囊的毛发。
[Abstract]:Preface ABO blood group is the first human blood genetic marker system. In 1990, ABO gene nucleic acid sequence was first reported by ABO et al. The differences in the base sequence of the alleles determining the phenotype of ABO were determined, which is the genetic basis for the detection of ABO genotypes at the DNA level. The ABO genotyping techniques used at present can be divided into amplified restriction fragment length polymorphism (PCR-RFLP) and single strand conformation polymorphism (SSCP) (SSCP) direct sequencing. Allele specific amplification method (ASA), allele specific primer consumption method (CASPA) and other techniques. Allele-specific amplification is one of the most widely used ABO genotyping methods in the field of forensic physical evidence testing and identification. This method is sensitive, simple and rapid. The allele-specific primer consumption (CASPA) method selected in this study is a new ABO genotyping method based on ASA recently. The advantage of this method is: 1. The method is simpler, faster and more sensitive because of the detection of remaining fluorescent labeled primers by capillary electrophoresis. Because the length of the allele-specific primers was much shorter than that of the amplified products, the separation time of the allele-specific primers was shortened by 3%. This method is suitable for ABO genotyping of biodegradable samples because of the amplified residual primers. Based on the allele-specific primer consumption method reported by Watanabe et al., the primer design was improved, and the applicability of common biological evidence in forensic medicine was studied. The results are reported as follows: the experimental materials were labeled with fluorescent dye 6-FAM and HEX, respectively. Six allele-specific primers with different lengths and one common primer with no marker. DNA extract of known ABO genotypes. 146 blood spot samples from unrelated individuals of Han nationality in China. Mixed spot, cigarette butt, sweat spot, muscle, cartilage, femur, hair with hair follicle.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：D919.1

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