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PubMed1974

Evaluation of micro complement fixation tests for antibodies against group A streptococcal M and M-associated antigens in rabbit and human sera.

Variables involved in micro complement fixation (CF) tests for type-specific (TSM) and non-type-specific (NTSM) antibodies against preparations of streptococcal M protein were studied. Sera of rabbits immunized with purified M protein which contained high titers of anti-TSM and low titers of anti-NTSM antibodies reacted type specifically in the CF tests when relatively low concentrations of M protein were employed. In contrast to these artificially induced TSM antibody responses, the sera of rheumatic fever patients demonstrated elevated CF antibody titers (1:80 to 1:320) in the presence of both high and low concentrations of M antigen. The CF test cannot distinguish TSM and NTSM antibodies when the latter predominate. The CF test can be employed with confidence for the detection of NTSM antibodies, provided that the test sera are known to lack type-specific antibodies against the M protein used in the test.

Animals↗
PubMed1975

New complement-fixation test for the human reovirus-like agent of infantile gastroenteritis. Nebraska calf diarrhea virus used as antigen.

A complement-fixation (C.F.) test for the human reovirus-like agent of infantile gastroenteritis has been developed using the serologically related Nebraska calf diarrhoea virus (N.C.D.V.) as antigen. Most infants and children who shed the agent in stools and/or who demonstrated serological (C.F.) evidence of infection with a reovirus-like-particle-positive human stool-filtrate C.F. antigen also demonstrated serological evidence of infection when a concentrated N.C.D.V. preparation was employed AS C.F. antigen. The N.C.D.V., which was previously shown to be related to the human reovirus-like agent, was found to be related antigenically to the epizootic diarrhoea of infant mice (E.D.I.M.) virus also. Studies on the prevalence of C.F. antibody in sera from infants and young children revealed a pattern of rapid acquisition of antibody to both the human reovirus-like agent and the N.C.D.V. as over 80 percent of these individuals possessed antibody to each agent by 36 months of age. A strong positive association was found in the results obtained with the two antigens. The ready availability of cell-culture grown N.C.D.V., and its ability to serve as a "substitute" C.F. antigen for the human reovirus-like agent, should enable the serodiagnosis of many cases of disease due to the human agent and facilitate seroepidemiological studies of such infections. In addition, the observation that a large proportion of individuals infected with the human reovirus-like agent develop serological evidence of infection not only to the human agent but to the calf agent as well may have important implications in the immunoprophylaxis of disease caused by the human reovirus-like agent.

Animals↗
PubMed1991

Comparison of seroreactivity of rams with brucellosis in a complement fixation test, whole cell ELISA and by immunoblotting.

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 kDa.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗