Forensic Applications of ELISA

1. 制备了针对ABO和MN抗原、糖蛋白A和B、IgG同种异体G3M G和T、a2 -半糖蛋白（A2SGP）和百草枯的单克隆抗体（MoAbs）用于elisa。以ALB、IgG、A2M、HP、TF、GC、A2HS、BF、HF为指示蛋白，采用抑制elisa法对人血进行鉴定。以抗尿黏液样抗体为捕获抗体，采用ELISA法对人尿污渍进行了最好的鉴定。体液ABO分组的自动化和数据处理是通过一个反应单元的组装来实现的，该反应单元包含一个涂有染色衍生的、洗涤剂溶解的抗原的板和一个自动读取单元。采用MoAbs对G3M G和G3M t进行检测，建立了一种简便的点免疫结合（dot-ELISA）基因分型方法，该方法适用于日常法医案件工作。建立了百草枯竞争性酶联免疫吸附测定法，可测定1 ~ 10 ng/ml的农药残留量。唾液、精液和混合染色的血液可以通过使用相应抗体捕获elisa进行不同ABO分组。特别是，自制的MoAbs对携带ab的A2SGR特异性捕获相应的蛋白质，从而能够对混合阴道液的精液进行正确的ABO分组。因此，这种方法在强奸案件中显得很有用。针对IgG异型GlM F和GlM X的单克隆抗体的生产正在进行中，因为日本人的常规转基因分型需要四种转基因单克隆。

1. Monoclonal antibodies (MoAbs) to ABO and MN antigens, glycophorin A and B, IgG allotypes G3M G and T, A2-seminoglycoprotein (A2SGP), and paraquat were produced for use in ELISAs.2. Human blood could be identified by inhibition-ELISAs using ALB, IgG, A2M, HP, TF, GC, A2HS, BF, and HF as indicator proteins. Human urinary stains were best identified by a capture ELISA using anti-uromucoid as the capture antibody.3. Automation and data processing for ABO grouping of body fluids were effected by assembly of a reaction unit containing a plate coated with stain-derived, detergent-solubilized antigen, and an automatic reading unit.4. A simple dot-immunobinding (dot-ELISA) method for GM typing was developed using MoAbs to G3M G and G3M T. The method is well suited for routine forensic casework.5. A competitive ELISA for paraquat was developed which enabled determination of 1-10 ng/ml of the pesticide.6. Saliva, semen, and blood in mixed stains could be differentially ABO grouped by capture ELISAs using corresponding antibodies. In particular, home-made MoAbs to AB-carrying A2SGR specifically captured the corresponding protein, thereby enabling correct ABO grouping of semen mixed with vaginal fluids. Thus the method appears useful in rape casework.7. Production of MoAbs to IgG allotypes GlM F and GlM X is in progress, since four GM monoclonals are required for routine GM typing of Japanese.