title: Development of an Indirect ELISA Using Different Fragments of Recombinant Ncgra7 for Detection of Neospora caninum Infection in Cattle and Water Buffalo.
abstract: BACKGROUND
  Dense granules are immunodominant proteins for the standardization of immunodiagnostic procedures to detect neosporosis. In the presented study different fragment of a dense-granule protein was evaluated for serodiagnosis of Neospora caninum in cattle and water buffalo.
  
  
  METHODS
  NcGRA7, from N. caninum tachyzoites was amplified. PCR product and pMAL-c2X plasmid were digested with EcoR1 restriction enzyme and expressed in Escherichia coli to evaluate its competence for detection of anti- N. caninum antibodies with ELISA in comparison with commercial IDEXX ELISA. Furthermore, 230 sera of presumably healthy cattle and water buffaloes (108 cattle and 122 water buffaloes) were analyzed by both tests to determine the agreement of these two procedures.
  
  
  RESULTS
  Sensitivities and specificities of NcGRA7-based ELISA were 94.64% and 90.38% respectively using sera of cattle, but were 98.57% and 86.54% in the case of buffaloes respectively. A good correlation between the results of IDEXX ELISA and ELISA based on recombinant NcGRA7 for detecting N. caninum antibodies was appeared. Analyzing by Mc Nemar's showed that NcGRA7-based ELISA has acceptable capability to differentiate the positive results in comparison with IDEXX ELISA.
  
  
  CONCLUSIONS
  NcGRA7-based ELISA considering utilized new fragment of genomic DNA is a good tool for serodiagnosis of anti- N. caninum antibodies for screening and epidemiological purposes on cattle herd and water buffaloes as well.
authors.affiliation: Dept. of Pathobiology, Veterinary Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Hamidinejat
authors.affiliation: Dept. of Pathobiology, Veterinary Faculty, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
authors.forename: Massoud Reza
authors.initials: MR
authors.lastname: Seifi Abad Shapouri
authors.affiliation: Razi Vaccine and Serum Research Institute, Shiraz Branch, Shiraz, Iran.
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Namavari
authors.affiliation: Dept. of Parasitology, Veterinary Faculty, University of Tehran, Tehran, Iran.
authors.forename: Parviz
authors.initials: P
authors.lastname: Shayan
authors.affiliation: Dept. of Parasitology, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
authors.forename: Marzieh
authors.initials: M
authors.lastname: Kefayat
keywords: Cattle; ELISA; Iran; NcGRA7; Neospora caninum; Recombinant antigen; Water buffalo