Development of a polymerase chain reaction assay for the diagnosis of neosporosis using the Neospora caninum 14-3-3 gene |
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Affiliation: | 1. ANSES, Laboratoire de santé animale de Maisons-Alfort, UMR BIPAR, Université Paris-Est, Maisons-Alfort, France;2. National Reference Laboratory for Toxoplasmosis, Institute for Medical Research, University of Belgrade, Dr. Subotića 4, P.O. Box 102, Belgrade 11129, Serbia;3. Ecole Nationale Vétérinaire d''Alfort, UMR BIPAR, Université Paris-Est, Maisons-Alfort, France;4. Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Naples, Italy;1. Transboundary Animal Diseases Research Center, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto 1-21-24, Kagoshima, Japan;2. Department of Veterinary Parasitological Diseases, Faculty of Applied Biological Sciences, Gifu University, Yanagido 1-1, Gifu, Japan;3. Department of Veterinary Clinical Theriogenology, Faculty of Applied Biological Sciences, Gifu University, Yanagido 1-1, Gifu, Japan;4. Yamatokohgen Animal Medical Clinic, Habikino, Osaka, Japan |
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Abstract: | Neospora caninum is a recently described apicomplexan parasite which causes neuromuscular disease in dogs, and abortion and neonatal morbidity in cattle, sheep and horses. Morphological similarites and serological cross-reactivity between N. caninum and the closely related parasite Toxoplasma gondii, have resulted in the frequent misdiagnosis of neosporosis as toxoplasmosis. This report describes the isolation and characterization of an N. caninum cDNA clone encoding a 14-3-3 protein homologue. The 14-3-3 proteins are a class of proteins which show a high degree of amino acid sequence conservation across several eukaryotic taxa. Using less conserved regions of the N. caninum cDNA clone, nested primers were designed for the amplification of a 614-bp N. caninum DNA fragment by the polymerase chain reaction (PCR). The DNA fragment was amplified from N. caninum genomic DNA, but not from T. gondii, Sarcocystis muris, Sarcocystis tenella, or Sarcocystis cruzi genomic DNA. Additionally, the fragment was amplified from DNA prepared from the brains of N. caninum-infected mice, but not from the brain of a mouse infected with T. gondii. These results suggest that this PCR assay may be useful for the diagnosis of neosporosis. |
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