Whole-bacterial cell enzyme-linked immunosorbent assay for cell-bound Moraxella bovis pili.
Infectious bovine keratoconjunctivitis (IBK), caused by Moraxella bovis, is a disease of major importance in cattle industry. M. bovis has several virulence factors among which pili are crucial antigen for the protective capacity of vaccines against this disease. The production of vaccines against IBK therefore requires a reliable technique for cellular piliation level assessment on cells to be included as vaccine components. In this study we describe a specific whole-bacterial cell enzyme-linked immunosorbent assay (bact-ELISA) capable of detecting pili antigen on M. bovis cell surface. A sequential competitive bact-ELISA was developed using highly piliated M. bovis cells as antigen. Samples to be analyzed were allowed to react with anti-pilus serum prior to incubation in wells coated with piliated cells of M. bovis. This assay proved useful for the rapid, sensitive and reproducible evaluation of piliation on M. bovis cells, and represents an important tool for cellular piliation monitoring daburing M. bovis cells production in stirred bioreactors.
Prieto
Claudia I
CI
Facultad de Ciencias Exactas, Centro de Investigación y Desarrollo en Fermentaciones Industriales, Universidad Nacional de La Plata (UNLP), Calle 47 y 115, 1900, La Plata, Argentina.
Rodriguez
María E
ME
Bosch
Alejandra
A
Chirdo
Fernando G
FG
Yantorno
Osvaldo M
OM
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial; D015153:Blotting, Western / Q000662:veterinary; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000276:immunology / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005260:Female; D010861:Fimbriae, Bacterial / Q000276:immunology* / Q000648:ultrastructure; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D007639:Keratoconjunctivitis, Infectious / Q000276:immunology / Q000382:microbiology*; D008854:Microscopy, Electron / Q000662:veterinary; D016955:Moraxella bovis / Q000276:immunology*; D011817:Rabbits; D013047:Specific Pathogen-Free Organisms
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial