Padronização da imunohistoquímica para detecção de clostridium chauvoei e clostridium septicum e comparação com a imunofluorescência direta

Abstract

Standardized the technique of immunohistochemistry using the complex streptavidin-biotin immunoenzymatic marked (LSAB) to detect Clostridium septicum and Clostridium chauvoei and compared it with the technique of direct immunofluorescence (DIF), using different tissues of guinea pigs inoculated with chauvoesi Clostridium, Clostridium septicum, Clostridium sordellii, Clostridium novyi type A and Clostridium chauvoei and anti-Clostridium septcum were produced in rabbits and purified by DEAE-cellulose hill. Portions of imonoglobunas were conjugated with FITC and the remainder was used in the LSAB technique. Conjugates anto-and anti-chauvoei Clostridium Clostridium septicum showed titers of 256 and 512, respectively, while the primary anti-Clostridium chuvoei and anti-Clostridium septicum were standardized at a dilution of 1:2000 / 1:4000 and 15 minutes / 40 minutes of incubation, respectively. There were no cross-reactivity between antibodies evaluated and none of the other species of clostridia used. Chauvoei Clostridium septicum and Clostridium were detected in all sections and tissue impressions by LSAV techniques and IFD. LSAB technique developed in this study was efficient to detect Clostridium septicum and Clostridium chauvoei in tissues of guinea pigs fixed in formalin and embedded in paraffin, and had 100% sensitivity and specificity of DIF