02665naa a2200337 a 450000100080000000500110000800800410001902400380006010000160009824501520011426000090026652016730027565000160194865000220196465000380198665000190202465000150204365000130205865000390207165000190211065000210212965000140215065000160216465000120218065000130219265000100220565300220221570000130223770000170225077300600226715203972023-05-02       1991    bl uuuu     u00u1 u    #d7 a10.1016/0378-1135(91)90022-82DOI1 aCHIN, J. C.  aComparison of seroreactivity of rams with brucellosis in a complement fixation test, whole cell ELISA and by immunoblotting.h[electronic resource]  c1991  aAbstract: In rams with ovine brucellosis, a high degree of serological correlation exists between the complement fixation (CF) test which utilises antigen extracted from bacteria with hot saline, and the ELISA reactivity using methanol-fixed Brucella ovis as the assay reagent. Since the whole cell ELISA (CELISA) detects mainly antibodies against surface antigens of B. ovis, it was concluded that the similar findings of the two serological tests is due in part to the presence of membrane antigens in the CF test antigen following hot saline extraction of intact bacteria. Immunoblots with pooled sera representing different CF titres confirmed that the major immunoreactive antigens of B. ovis were located in four zones: alpha, beta, gamma 1 and 2 with corresponding apparent molecular masses of 55 and 60 kDa; 27 and 29 kDa; 18.5-20 kDa and 17-18 kDa, respectively. These zones of reactivity were consistently present in immunoblots when assayed against different B. ovis isolates even though Coomassie brilliant blue staining of SDS-PAGE gels revealed some differences in polypeptide banding patterns. However, these intensely-stained CBB bands located at 38 and 40 kDa which distinguished three of the seven B. ovis isolates were considerably less reactive in immunoblots compared to polypeptides that were located at positions equivalent to alpha, beta or gamma reactivities. Intensity of immunoblot reactivity against polypeptides located in the alpha, beta and gamma zones intensified with increasing CF titre. Sera with CF titres greater than 32 also tended to react against bands of higher apparent molecular masses located at 65, 70, 73, 78, 80 and 86 kD  aBrucellosis  aDielectrophoresis  aEnzyme-linked immunosorbent assay  aImmunoblotting  aImmunology  aPeptides  aPolyacrylamide gel electrophoresis  aSheep diseases  aSurface antigens  aBrucelose  aDiagnostico  aDoença  aEpidemia  aOvino  aBrucella analysis1 aPANO, B.1 aCARRIGAN, M.  tVeterinary Microbiologygv. 26, n. 3, p. 291-299, 1991.