OBTENTION OF A SPECIFIC CONJUGATE FOR THE RAPID DETECTION OF VIBRIO PARAHAEMOLYTICUS

BARBINI NB; REAL S; ALMEIDA C; DIGENARO MS; MICALIZZI B; STEFANINI DE GUZMÁN AM

Congreso; XX Reunión de la Sociedad de Biología de Cuyo; 2002

Sociedad de Biología de Cuyo

Vibrio parahaemolyticus causes gastroenteritis associated with theconsumption of seawater fish and shellfish. The purpose was toobtain a V. parahaemolyticus?specific fluorescein isothiociante(FITC) conjugated IgG, in order to be used for the rapid detectionof this bacterium in clinical and food samples. Three rabbits wereimmunized with a formalised V. parahaemolyticus ATCC 17802.The sera were titled by ELISA and agglutination. The IgG waspurified using protein A-Sepharose affinity chromatography andelution with glycine buffer pH 3. The purified antibody was labeledwith 5 mg/ml FITC in DMSO (20µl/mg of antibody). After removingunbound FITC by dialysis, protein concentration, FITC/antibodyratio was calculated. The conjugate was tested by directimmunofluorescence (IFD). The serum title was 1/800 by ELISAand 1/64 by agglutination. The FITC/protein ratio was 4.12. Theconjugate reacted with V. parahaemolyticus. The conjugate obtainedis optimal for IFD and can be used in the rapid detection of thisbacterium