Use of detergent extracts of Brucella abortus RB51 to detect serologic responses in RB51-vaccinated cattle.
Serologic responses to the newly introduced rough Brucella abortus vaccine strain RB51 have been determined in a dot-blot format using gamma-irradiated RB51 cells as the antigen. Because gamma-irradiated cells are not easily prepared and the signal from cells was not always reliable, an alternative antigen was sought. Detergent extracts of B. abortus RB51 were prepared using zwittergent 3-14, Triton X-100, and sodium dodecyl sulfate (SDS) and examined in a dot-blot format. Zwittergent 3-14 extracts and gamma-irradiated RB51 cells gave the same titers. Unlike gamma-irradiated RB51 cells, zwittergent 3-14 extracts produced signals consistently, and the signals were easily interpreted. Triton X-100 extracts interfered with signal development, and SDS extracts resulted in a high background signal. Western blot analyses revealed several outer membrane proteins in the zwittergent 3-14 extract. The major antigens in the extract had apparent molecular weights of <20,000.
Halling
S M
SM
Bacterial Diseases of Livestock Unit, National Animal Disease Center, Agricultural Research Service, USDA, Ames, IA 50010-0070, USA.
Koster
N A
NA
D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis*; D001428:Bacterial Vaccines; D015153:Blotting, Western; D002003:Brucella abortus / Q000276:immunology* / Q000302:isolation & purification / Q000472:pathogenicity; D002007:Brucellosis, Bovine / Q000175:diagnosis* / Q000276:immunology; D002417:Cattle; D003902:Detergents / Q000737:chemistry*; D008970:Molecular Weight; D017830:Octoxynol / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012698:Serologic Tests / Q000662:veterinary; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D013048:Specimen Handling; D014611:Vaccination / Q000662:veterinary*
D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D003902:Detergents; D000644:Quaternary Ammonium Compounds; C048256:zwittergent 3-14; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol