Rapid and sensitive identification of Neospora caninum by in vitro amplification of the internal transcribed spacer 1.
Neospora caninum and N. caninum-like organisms are cyst-forming coccidian parasites known to cause neuromuscular disorders in dogs and abortion in cattle. In this article we report on the use of the polymerase chain reaction (PCR) for the detection of DNA from N. caninum. After determining the sequence of the internal transcribed spacer 1 (ITS1) of N. caninum and Toxoplasma gondii, and part of the sequences for 4 species of Sarcocystis, we designed a primer set for the amplification of a 279-base-pair fragment of ITS1 from N. caninum. The PCR system made possible the specific detection of 5 N. caninum organisms and no amplification was observed from any of the other cyst-forming coccidia tested, including the closely related T. gondii. Furthermore, we were also able to demonstrate the presence of N. caninum in brain and lung tissue samples from experimentally infected mice. Our data also link the 5.8S rRNA gene for T. gondii and N. caninum to the 16S-like rRNA gene, within the rDNA unit.
Holmdahl
O J
OJ
Department of Parasitology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Mattsson
J G
JG
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000469:parasitology; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics*; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics*; D005260:Female; D008168:Lung / Q000469:parasitology; D051379:Mice; D008969:Molecular Sequence Data; D018688:Neospora / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics; D012522:Sarcocystis / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D014122:Toxoplasma / Q000235:genetics
D017931:DNA Primers; D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012340:RNA, Ribosomal, 5.8S