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- Title
Analytical sensitivity of a multiplex quantitative PCR for Toxoplasma gondii and Neospora caninum.
- Authors
Truong, Marcus; Šlapeta, Jan
- Abstract
Cyst-forming coccidia, Toxoplasma gondii and Neospora caninum, are recognised as important causes of animal disease. Molecular diagnostics based on the presence of DNA in animal tissue are required to specifically detect T. gondii and N. caninum while achieving high levels of analytical sensitivity. We optimised available single-plex probe base qPCR assays into a multiplexed qPCR panel to detect cyst-forming coccidia, i.e. T. gondii and N. caninum. The T. gondii assay is based on a 529-bp repetitive (REP) element and the N. caninum assay on the NC5 repetitive region. Using target sequence synthetic DNA, the limit of detection (LOD) was determined to be 100 copies, that is less than a single tachyzoite of either T. gondii or N. caninum. The T. gondii and N. caninum multiplexed qPCR assay optimised in this study can be used to effectively detect parasite DNA for diagnostic purposes in animal tissue.
- Subjects
NEOSPORA caninum; TOXOPLASMA gondii; ARTIFICIAL chromosomes; ANIMAL diseases; POLYMERASE chain reaction; COCCIDIA; MOLECULAR diagnosis
- Publication
Parasitology Research, 2023, Vol 122, Issue 4, p1043
- ISSN
0932-0113
- Publication type
Article
- DOI
10.1007/s00436-023-07796-5