Use of hydrophobic cloths as antibody adsorbents for enzyme immunoassay: detection of Brucella antigens.
A cloth-ELISA (C-ELISA) antigen capture assay for the detection of Brucella melitensis and B. abortus was developed. Segments (6-mm squares) of hydrophobic polyester cloth coated with diluted serum from a B. abortus-infected cow were incubated with saline suspensions of heat-killed Brucella cells, or with cultures of bovine or sheep blood or bovine tissue homogenates that had been inoculated with B. abortus or B. melitensis added to trypticase soy broth (TSB) and incubated for 2-3 days. The captured antigen was detected by a bovine anti-Brucella antibody-horseradish peroxidase conjugate system. The enrichment culture technique detected as few as three Brucella colony-forming units (c.f.u.) in 0.5 ml of bovine blood and was positive in cultures in which the Brucella concentration had reached 3 X 10(6) c.f.u. ml-1 (after 2 or 3 days incubation). The combined enrichment-cloth-ELISA method gave complete correlation with cultural isolation and results were available 3 days before colonies appeared in conventional culture. Hydrophobic cloths have potential use in diagnostic procedures since they provide simple, rapid and economical assays.
Blais
B W
BW
Department of Biology, Carleton University, Ottawa, Ont., Canada.
Yamazaki
H
H
Rigby
C E
CE
D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis*; D002002:Brucella / Q000276:immunology*; D002003:Brucella abortus / Q000276:immunology*; D002007:Brucellosis, Bovine / Q000276:immunology*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007106:Immune Sera / Q000276:immunology; D013997:Time Factors
D000942:Antigens, Bacterial; D007106:Immune Sera