Forensic Applications of ELISA

1.制备了抗ABO和MN抗原、血糖蛋白A和B、同种异型G3M G和T、A2-精糖蛋白(A2SGP)和百草枯的单抗(MoAbs)。以ALB、Ig G、A2M、Hp、Tf、Gc、A2HS、BF、Hf为指示蛋白，采用抑制ELISAs法对人血进行鉴定。以抗尾粘液为捕获性抗体的捕捉法检测人尿斑效果最好。体液ABO分组的自动化和数据处理是通过组装一个反应单元实现的，该反应单元包含一个涂有染料衍生的洗涤剂溶解抗原的平板和一个自动读取单元。利用MoAbsG3MG和G3MT建立了一种简便的GM型斑点免疫结合(Dot-ELISA)方法，该方法适用于常规法医学案件。建立了百草枯的竞争酶联免疫吸附测定方法，可检测到1-10 ng/ml的农药。混合斑块中的唾液、精液和血液可以通过使用相应抗体捕获ELISA来区分ABO分组。特别是，自制的MoAbto AB携带的A2SGR特异性地捕获了相应的蛋白质，从而使混合了阴道液的精液能够正确地进行ABO分组。因此，该方法在强奸案中是有用的。由于日本人的常规GM分型需要四个GM单克隆，因此针对GLm F和GLm X的免疫球蛋白同种异型的MoAbs的生产正在进行中。

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.