title: Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice.
abstract: Toxoplasma gondii is a zoonotic parasite with a world-wide distribution. House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to improve the diagnosis, we tested a novel protocol for T. gondii detection in mice and compared this technique to a standard PCR-based protocol using a commercial kit for DNA isolation. Efficacy of magnetic capture for isolation of T. gondii DNA from whole host brains was tested in brain samples of laboratory mice spiked with 1 up to 10(4) tachyzoites. Real-time PCR revealed that even 1-5 tachyzoites can be detected after magnetic capture. Also this method is suitable to quantify parasite numbers in mouse brains with more than 10 tachyzoite equivalents. To assess the two techniques in wild mice, we employed a dataset consisting of 243 individuals. The prevalence of T. gondii detected by magnetic capture and qPCR and by commercial isolation and PCR was 1.2% and 0%, respectively. The magnetic capture and quantitative PCR seems to be a highly sensitive and specific diagnostic method for both laboratory research and wild population surveys.
authors.affiliation: Department of Pathology and Parasitology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic. Electronic address: yanniska@seznam.cz.
authors.forename: Jana
authors.initials: J
authors.lastname: Juránková
authors.affiliation: Department of Pathology and Parasitology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic; Central European Institute of Technology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic.
authors.forename: Lada
authors.initials: L
authors.lastname: Hůrková-Hofmannová
authors.affiliation: Department of Bacteriology, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Volf
authors.affiliation: Department of Biology and Wildlife Diseases, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic.
authors.forename: Vojtech
authors.initials: V
authors.lastname: Baláž
authors.affiliation: Institute of Vertebrate Biology, Academy of Sciences of the Czech Republic, Brno and Studenec, Czech Republic.
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Piálek
mesh_terms: D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis / Q000302:isolation & purification*; D005260:Female; D005821:Genetic Techniques / Q000592:standards*; D057230:Limit of Detection; D060328:Magnetic Phenomena; D051379:Mice; D016133:Polymerase Chain Reaction / Q000592:standards; D014122:Toxoplasma / Q000235:genetics*; D014124:Toxoplasmosis, Animal / Q000175:diagnosis*
chemical_list: D016054:DNA, Protozoan
keywords: House mice; Magnetic capture; Quantitative PCR; Toxoplasma gondii