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Detection of Neospora caninum in dog organs using real time PCR systems

Ghalmi, Farida, China, Bernard, Kaidi, Rachid, Daube, Georges, Losson, Bertrand
Veterinary parasitology 2008 v.155 no.1-2 pp. 161-167
dogs, dog diseases, Neospora caninum, animal pathogens, neosporosis, disease detection, animal tissues, paresis, polymerase chain reaction, microbial genetics, nucleotide sequences, liver, spleen, disease prevalence, fluorescent antibody technique, accuracy, seroprevalence
Neospora caninum is a parasite responsible for paresis in dogs. The dog can harbour enkysted parasites in several organs. The detection of N. caninum was performed using 3 different real time PCR systems all amplifying the NC5 DNA region. One system was based on Sybr®green, one on Plexor[trade mark sign] technology and the last on Taqman® probe. Comparison of the three methods indicated that the detection limit was 1 equivalent genome on pure DNA but that this detection limit increased in the presence of foreign DNA using the Sybrgreen and Plexor systems. Therefore, the Taqman system was chosen to detect N. caninum in liver and spleen of naturally infected dogs. The overall prevalence was 32.2%. Comparison between PCR results and serological results using IFAT showed that among the 28 PCR positive dogs only 9 were seropositive and that 8 seropositive dogs were PCR negative. Therefore serology can underestimate the real carriage in dogs. However, PCR methods must be improved in terms of sensitivity and inhibition problems.