title: Monoclonal antibodies directed against the outer membrane protein of Bordetella avium.
abstract: Bordetella avium is the etiologic agent of coryza and rhinotracheitis in poultry. This respiratory disease is responsible for substantial economic losses in the poultry industry. Monoclonal antibodies (MAbs) were produced against the outer membrane proteins (OMPs) of B. avium isolated from diseased chickens. BALB/c mice were immunized with the extracted B. avium OMPs. Then the splenocytes from immunized mice and SP2/0 myeloma cells were fused using PEG 4000. Three stable hybridoma clones (designated as 3G₁₀, 4A₃, and 4E₈) were produced via indirect ELISA and three rounds of subcloning. The MAbs were classified as IgG1, and can recognize the 58  kDa OMP band by Western blot assays. No MAb cross-reactivity with chicken Proteus mirabilis, Escherichia coli, and Salmonella was observed. A double antibody sandwich ELISA (DAS-ELISA) was developed using the rabbit polyclonal antibodies as the capture antibody and MAb 4A₃ as the detection antibody. Under the DAS-ELISA, the minimum detectable concentration of B. avium was 1 × 10(4) CFU/mL, and no cross-reactivity occurred with chicken Proteus mirabilis, Escherichia coli, and Salmonella. Results showed that the DAS-ELISA has good sensitivity and specificity. Clinical application showed the DAS-ELISA was more sensitive than the plate agglutination test. This study may be used to develop a quick and specific diagnostic kit, analyze epitopes, and establish systems for typing B. avium.
authors.affiliation: Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Technology, Shandong Agricultural University, Shandong Taian, PR China.
authors.forename: Guanhua
authors.initials: G
authors.lastname: Liu
authors.forename: Manfei
authors.initials: M
authors.lastname: Liang
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zuo
authors.forename: Xue
authors.initials: X
authors.lastname: Zhao
authors.forename: Fanxia
authors.initials: F
authors.lastname: Guo
authors.forename: Shifa
authors.initials: S
authors.lastname: Yang
authors.forename: Ruiliang
authors.initials: R
authors.lastname: Zhu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D000917:Antibody Formation; D000918:Antibody Specificity; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D001885:Bordetella Infections / Q000097:blood / Q000276:immunology* / Q000382:microbiology; D042482:Bordetella avium / Q000276:immunology*; D002645:Chickens / Q000097:blood / Q000276:immunology* / Q000382:microbiology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins
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