Memórias do Instituto Oswaldo Cruz | |
The use of dacron plates for DOT enzyme-linked immunosorbent assay (DOT-ELISA) | |
Silvia M. L. Montenegro2  Alzira M. P. De Almeida2  Alexandre B. De Carvalho2  Luiz B. De Carvalho Júnior1  | |
[1] ,FIOCRUZ Centro de Pesquisas Aggeu Magalhaes Recife,Brasil | |
关键词: dacron; dot-ELISA; Yersinia pestis; | |
DOI : 10.1590/S0074-02761991000400016 | |
来源: SciELO | |
【 摘 要 】
Dacron (polyethylenetherephthalate) is proposed as a matrix for dot-ELISA procedures, as an alternative to nitrocellulose. Plates of dacron were partially hydrazinolyzed and hydrazide groups introduced were converted to azide groups. The derivative dacron-antigen was covalently linked on to the plates through these azide groups. The derivative dacron-antigen was exaustively washed according to CROOK and antigen was still fixed onto the plates. Protein F1A purified from Yersinia pestis was used as a model. Triration of sera from immunized and non immunized rabbits against this protein was carried out by employing the dot-ELISA method. No significant difference was observed using dacron-antigen and nitrocellulose-antigen preparations. However, both procedures showed to have a significant better performancein comparasion with the passive hemagglutination method. The specificity and reproductibility of the dot-ELISA assay using both preparations showed a similar behaviour. Nitrocellulose preparation was stable at 4ºC, 28ºC and -20ºC for 90 days, whereas the dacron-antigen derivative was stable only when stored at 4ºC. Dacron-antigen derivative could be re-used when the spot developing was proceeded using 4-chloro-1-naphtol as substrate.
【 授权许可】
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