聚乙烯醇戊二醛在鼠疫酶联免疫吸附测定中作为固相的应用。
The use of polyvinyl alcohol glutaraldehyde as solid-phase in ELISA for plague.
作者信息
Araujo A M, Petribú A T, Sales Barbosa G H, Diniz J R, de Almeida A M, Mendes Azevedo W, Malagueño E, Carvalho Júnior L B
机构信息
Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco, Recife, Brasil.
出版信息
Mem Inst Oswaldo Cruz. 1996 Mar-Apr;91(2):195-8. doi: 10.1590/s0074-02761996000200014.
Discs of polyvinyl alcohol cross-linked with glutaraldehyde were synthesized under acid catalysis (H2SO4). Then, the antigen F1 purified from Yersinia pestis was covalently linked to this modified polymer. Afterwards, an enzyme-linked immunosorbent assay (ELISA) was established for the diagnosis of plague in rabbit and human. The best conditions for the method were achieved by using 1.3 micrograms of F1 prepared in 0.067 M phosphate buffer, pH 7.2, containing 1 M NaCl (PBS); anti-IgG peroxidase conjugate diluted 6,000 times and as a blocking agent 3% w/v skim milk in PBS. The titration of positive rabbit serum according to this procedure detected antibody concentrations up to 1:12,800 times. The present method, the conventional ELISA and passive haemagglutination assay are compared.
在酸催化（硫酸）条件下合成了与戊二醛交联的聚乙烯醇圆盘。然后，将从鼠疫耶尔森氏菌中纯化的抗原F1与这种改性聚合物共价连接。之后，建立了一种酶联免疫吸附测定法（ELISA）用于兔和人的鼠疫诊断。该方法的最佳条件是使用在含有1M氯化钠（PBS）的pH 7.2的0.067M磷酸盐缓冲液中制备的1.3微克F1；抗IgG过氧化物酶缀合物稀释6000倍，并用PBS中3% w/v脱脂乳作为封闭剂。按照此程序对阳性兔血清进行滴定，检测到的抗体浓度高达1:12800倍。对本方法、传统ELISA和被动血凝试验进行了比较。
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