Iranian Journal of Parasitology 2015. 10(1):69-77.

Development of an Indirect ELISA Using Different Fragments of Recombinant Ncgra7 for Detection of Neospora caninum Infection in Cattle and Water Buffalo.
Hossein Hamidinejat, Massoud Reza Seifi Abad Shapouri, Mohammad Mehdi Namavari, Parviz Shayan, Marzieh Kefayat

Abstract


 

Background: Dense granules are immunodominant proteins for the standardi-zation of immunodiagnostic procedures to detect neosporosis. In the presented study different fragment of a dense-granule protein was evaluated for serodiagno-sis 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 ex-pressed in Escherichia coli to evaluate its competence for detection of anti- N. cani-num antibodies with ELISA in comparison with commercial IDEXX ELISA. Fur-thermore, 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 an-tibodies 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.

Conclusion:NcGRA7-based ELISA considering utilized new fragment of ge-nomic 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.


Keywords


Cattle; ELISA; Iran; NcGRA7; Neospora caninum; Recombinant antigen; Water buffalo

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