大肠杆菌O157志贺毒素1胶乳凝集试剂的研制

Latex Particle Reagent Prepared for of Escherichia coli O157 Shiga-Toxin 1

根据大肠杆菌933W志贺毒素1(STX1)基因的序列,设计合成寡核苷酸引物,以国内分离的大肠杆菌O157菌株94H的DNA为摸板,经PCR扩增志贺毒素1A亚基基因,扩增的基因克隆到原核表达载体pET-28a,在E.coliBL21中进行表达。用表达的STX1A产物免疫兔子,制备STX1A抗血清,从中提取IgG。合成胶乳,用提取的IgG与胶乳交连反应,研制出了敏感和简便的检测大肠杆菌O157STX1产生的胶乳检测试剂。

The sequence encoding of the mature protein of STX1 A of E.coli O157 strain 94H, isolated from some patient in China, was amplified by PCR, The primers were designed from STX1 toxin gene sequence of E. coli strain 933W. The amplified gene was cloned into prokaryotic expression vector pET-28a. Then the recombinant pET28a-stx 1A was transformed into host strain E. coli BL21 （DE3）. Expressed recombinant STX1 A was used to immunize rabbits to prepare anti-serum. The IgG fraction of the antiserum was cross-linked with the latex particle suspension to yield a final immunoglobulin. The described methods enabled a sensitive and simple assay of STX 1 A production.