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

By Silvia M. L. Montenegro, Alzira M. P. de Almeida, Alexandre B. de Carvalho and Luiz B. de Carvalho Júnior

Abstract

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 performance in 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

Topics: dacron, dot-ELISA, Yersinia pestis, Microbiology, QR1-502, Science, Q, DOAJ:Microbiology, DOAJ:Biology, DOAJ:Biology and Life Sciences, Arctic medicine. Tropical medicine, RC955-962, Special situations and conditions, RC952-1245, Internal medicine, RC31-1245, Medicine, R, DOAJ:Internal medicine, DOAJ:Medicine (General), DOAJ:Health Sciences
Publisher: Instituto Oswaldo Cruz, Ministério da Saúde
Year: 1991
DOI identifier: 10.1590/S0074-02761991000400016
OAI identifier: oai:doaj.org/article:a09cc6d530db41a380a7e19eab1271d8
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