The use of dacron plates for DOT enzyme-linked immunosorbent assay (DOT-ELISA)

Memórias do Instituto Oswaldo Cruz

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.

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