The use of dacron plates for dot enzyme-linked immunosorbent assay (dot-ELISA).
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. Titration 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 comparison 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 4 degrees C, 28 degrees C and -20 degrees C for 90 days, whereas the dacron-antigen derivative was stable only when stored at 4 degrees C. Dacron-antigen derivative could be re-used when the spot developing was proceeded using 4-chloro-1-naphtol as substrate.
Montenegro
S M
SM
Centro de Pesquisas Aggeu Magalhães, FIOCRUZ, Recife, Brasil.
de Almeida
A M
AM
de Carvalho
A B
AB
de Carvalho Júnior
L B
LB
D000818:Animals; D000942:Antigens, Bacterial / Q000302:isolation & purification; D003101:Collodion; D006386:Hemagglutination Tests; D015151:Immunoblotting / Q000295:instrumentation* / Q000379:methods; D011093:Polyethylene Terephthalates / Q000737:chemistry; D011817:Rabbits; D015010:Yersinia pestis / Q000276:immunology
D000942:Antigens, Bacterial; D011093:Polyethylene Terephthalates; D003101:Collodion