Fluorescent Antibody Techniques

Abstract

The diagnosis of early, mild, or atypical whooping cough is dependent upon identification of the etiologic agent. Although procedures for the isolation of Bordetella pertussis1 have been available for many years and are of proven diagnostic aid, they have not gained the wide usage they seem to deserve. A recognized limitation is the time needed: 3, more often 4, and occasionally more, days are required for a positive report. The possibility of using fluorescent antibody methods for rapid identification of B. pertussis in nasopharyngeal specimens is suggested by the promising results reported in recent studies of a number of bacterial infections. For example, Group A streptococci,2,3 enteropathogenic Escherichia coli,4 Haemophilus influenzae, Type B,5 Neisseria gonorrhoeae,6 Vibrio comma,7 Leptospira,8 and certain other bacterial agents have been demonstrated by means of this technique in specimens taken directly from the patient. The work of de Repentigny and Frappier,9 although not concerned with diagnosis, indicated that specific antigens of B. pertussis could be stained with dye-conjugated antibody. In a recent diagnostic study by Donaldson and Whitaker,10 31 of 36 nasopharyngeal specimens from infants with symptoms of clinical whooping cough were found positive with the fluorescent antibody technique ; cultural results were not given.In the present study, preliminary tests with an available pertussis antiserum conju¬ gated with fluorescein isothiocyanate showed brilliant fluorescence in slide preparations of several strains of B. pertussis; fluorescence was not observed with a number of cultures

FLUORESCENT ANTIBODY STAINING AND AGGLUTINATION REACTIONS IN BORDETELLA PERTUSSIS CULTURES

“…Reactions with unadsorbed Bordetella antisera. It has already been amply demonstrated by our own work (Kendrick et al, 1961), and by others (Repentigny and Frappier, 1956;Donaldson and Whitaker, 1960) that B. pertussis gives positive FA reactions with specific unadsorbed antiserum. Since cross-agglutination reactions are known to occur among the three Bordetella species, pertussis, parapertussis, and bronchiseptica, it was of interest to find out whether cross reaction could be demonstrated with the FA staining procedure.…”

Preservation of nasopharyngeal smears for fluorescent antibody detection of Bordetella pertussis

“…However, because of the decreased incidence of the disease, many laboratories do not maintain a fresh supply of Bordet-Gengou agar plates, and NP swabs are often transported to a central laboratory for processing. The viability of the pertussis organism is often lost after 2 h, even in a 1% Casamino Acids solution (7). Transport media which maintain viability for more than 2 h have not been adequately evaluated.…”

Culture and Fluorescent-Antibody Methods in Diagnosis of Whooping Cough

“…A total of 98 specimens were negative by both culture and FA staining. With respect to the FA method, Kendrick et al (1961) stressed the importance of a thorough knowledge of the organism under study, strict control of technical factors related to all steps in the procedure, and use of a suitable antiserum. Certain B. pertussis antisera produced in rabbits cause fluorescence with staphylococci, an observation also reported by other workers in studying other organisms.…”