Rapid and sensitive identification ofNeospora caninumbyin vitroamplification of the internal transcribed spacer 1

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Abstract
Summary: Neospora caninumandN. 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 fromN. caninum. After determining the sequence of the internal transcribed spacer 1 (ITSl) ofN. caninumandToxoplasma gondii, and part of the sequences for & species ofSarcocystis, we designed a primer set for the amplification of a 279-base-pair fragment of ITSl fromN. caninum. The PCR system made possible the specific detection of 5N. caninumorganisms and no amplification was observed from any of the other cyst-forming coccidia tested, including the closely relatedT. gondii. Furthermore, we were also able to demonstrate the presence ofN. caninumin brain and lung tissue samples from experimentally infected mice. Our data also link the 5-8S rRNA gene forT. gondiiandN. caninumto the 16S-like rRNA gene, within the rDNA unit.