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Brucella species (Serological and allergic skin tests (buffered Brucella…
Brucella species
Serological and allergic skin tests
buffered Brucella antigen tests, i.e. Rose Bengal test and
buffered plate agglutination test
complement fixation test,
enzyme-linked immunosorbent
assay (ELISA)
fluorescence polarisation assay,
Collection and culture of samples
Tissues: Samples are removed aseptically with sterile instruments. The tissue samples are prepared
by removal of extraneous material (e.g. fat), cut into small pieces, and macerated using a ‘Stomacher’
or tissue grinder with a small amount of sterile phosphate buffered saline (PBS), before being
inoculated on to solid media.
Vaginal discharge: A vaginal swab taken after abortion or parturition is an excellent source for the
recovery of Brucella and far less risky for the personnel than abortion material. The swab is then
streaked on to solid media.
Milk: Samples of milk must be collected cleanly after washing and drying the whole udder and
disinfecting the teats. It is essential that samples should contain milk from all quarters, and 10–20 ml of
milk should be taken from each teat.
Dairy products: Dairy products, such as cheeses, should be cultured on the media described above. As
these materials are likely to contain small numbers of organisms, enrichment culture is advised.
Serological tests
serum agglutination test (SAT)
complement fixation test (CFT) is diagnostically more specific than the SAT
enzyme-
linked immunosorbent assays (ELISAs)
fluorescence polarisation assay (FPA) are comparable with or
better than that of the CFT
Culture
Specimen
Blood or homogenized
spleen
Blue rods in short chains
surrounded by pink capsular
material (the M‟Fadyean reaction)
Polychrome methylene blue stained
smear
Capsule material is more abundant
if blood smear from recently dead
Basal media
Direct isolation and culture of Brucella
tryptose (or trypticase)–soy agar (TSA)
blood agar base (Oxoid) or Columbia agar (BioMérieux),
serum–dextrose agar (SDA) or glycerol dextrose
agar
Selective media
Appropriate
antibiotics are added to suppress the growth of organisms other than Brucella
Laboratory Diagnosis
Specimen
Ear/tail veins (sheep & goats)
Oedematous fluid (horses and
pigs)
Thin blood smear Technique
Peritoneal fluids (pigs)
Nucleic acid recognition methods
The PCR, including the real-time format, provides an additional means of detection and identification of Brucella sp.
Identification and typing
Laboratory Diagnosis for Brucella