title: Development and application of dot-enzyme-linked immunosorbent (dot-ELISA) assay for detection of Brucella melitensis and evaluation of the shedding pattern in infected goats.
abstract: Early and accurate diagnosis of Brucella melitensis is essential for the treatment and control of brucellosis both in animals and humans. The thrust for the development of a rapid diagnostic technique to overcome the limitations of conventional microbiological and serological tests brought about this investigation on the development and application of dot-ELISA for antigen and antibody detection in infected goats. Fifteen apparently healthy Boer aged 2-3 years which tested negative for brucellosis using PCR and ELISA, were grouped into A (10 goats infected intraocularly with 107CFU of B. melitensis) and B (5 goats) as control. Discharges (ocular, nasal, and vaginal) and blood were collected at days 3, 7, 10, 14, weekly until 42 post-infection (pi) for dot-ELISA, PCR, and RBPT. Dot-ELISA detected B. melitensis antigen and antibody in group A at day 3 and 7 pi, respectively with adequate sensitivity and specificity relative to PCR and RBPT. The bacteria shedding detected from discharges at day 3 pi in the nasal and ocular route with dot-ELISA. Group B were consistently negative. Values such as speed, simplicity, field adaptability, high sensitivity, and specificity make dot-ELISA a rapid and adequate technique for diagnosis of brucellosis in B. melitensis infected goats within few hours.
authors: a Department of Microbiology and Pathology , Universiti Putra Malaysia , Serdang , Malaysia.
authors: Opeyemi Mayowa
authors: OM
authors: Onilude
authors: a Department of Microbiology and Pathology , Universiti Putra Malaysia , Serdang , Malaysia.
authors: Sabri
authors: S
authors: Mohd Yusoff
authors: b Department of Veterinary Pathology , University of Ibadan , Ibadan , Nigeria.
authors: Benjamin Obukowho
authors: BO
authors: Emikpe
authors: a Department of Microbiology and Pathology , Universiti Putra Malaysia , Serdang , Malaysia.
authors: Polycarp
authors: P
authors: Tanko
authors: a Department of Microbiology and Pathology , Universiti Putra Malaysia , Serdang , Malaysia.
authors: Salisi M
authors: SM
authors: Shahrom
authors: c Institute of Marine Biotechnology , Universiti Malaysia Terengganu , Terengganu , Malaysia.
authors: Mohammed
authors: M
authors: Effendy
mesh_terms: D000818:Animals; D017347:Brucella melitensis / Q000276:immunology / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis / Q000276:immunology* / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006041:Goats / Q000382:microbiology*; D007164:Immunosorbents / Q000737:chemistry*
chemical_list: D007164:Immunosorbents
keywords: Brucella melitensis; diagnosis; dot-ELISA; shedding pattern