title: Immunoblot diagnosis of infection with Neospora caninum in cattle based on recombinant NcSAG4 Antigen.
abstract: The Neospora caninum (N. caninum) NcSAG4 gene was subcloned into a pET-28a (+) vector and successfully expressed in Escherichia coli as inclusion body, and confirmed by SDS-PAGE and Western blot using anti-His monoclonal antibody. The purified protein was then purified using Ni-NTA affinity chromatography column and recognized by positive serum from N. caninum-infected cattle. Immunoblot (IB) method based on purified recombinant NcSAG4 (rNcSAG4) antigen to detect antibodies against N. caninum in cattle was developed. Subsequently, both IB and ELISA kit were used to test sera (52) from naturally infected/uninfected cattle. Results showed that 50 and 48 out of 52 was positive for IB and ELISA kit, respectively, revealing that IB is more or at least as sensitive as ELISA when used for serodiagnosis of infected cattle.
authors.affiliation: Dipartimento di Produzioni Animali, Epidemiologia ed Ecologia, Università di Torino, No. 44 Via L. Da Vinci, Grugliasco, Torino, 10095, Italy. hujq0702@hotmail.com
authors.forename: Jinqiang
authors.initials: J
authors.lastname: Hu
authors.forename: Ezio
authors.initials: E
authors.lastname: Ferroglio
authors.forename: Anna
authors.initials: A
authors.lastname: Trisciuoglio
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000235:genetics / Q000302:isolation & purification; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003001:Cloning, Molecular; D003048:Coccidiosis / Q000175:diagnosis / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D015151:Immunoblotting / Q000379:methods; D018688:Neospora / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D010274:Parasitology / Q000379:methods*; D010957:Plasmids; D015800:Protozoan Proteins / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000627:therapeutic use; D012680:Sensitivity and Specificity
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins