Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice |
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| Institution: | 1. Department of Pathology and Parasitology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic;2. Central European Institute of Technology, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic;3. Department of Bacteriology, Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic;4. Department of Biology and Wildlife Diseases, University of Veterinary and Pharmaceutical Sciences, Palackého 1-3, 612 42 Brno, Czech Republic;5. Institute of Vertebrate Biology, Academy of Sciences of the Czech Republic, Brno and Studenec, Czech Republic;1. Department of Biological Sciences, Boise State University MS-1515, 1910 University Avenue, Boise, ID 83725-1515, USA;2. Department of Zoology, Comenius University, Mlynská dolina B-1, Bratislava SK-84215, Slovak Republic;1. Laboratory of Protozoology, Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, China;2. The Key Laboratory of Biomedical Information Engineering, Ministry of Education, Department of Biology and Engineering, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an 710049, China;3. Zoology Department, King Saud University, Riyadh 11451, Saudi Arabia;4. Laboratory of Protozoology, Key Laboratory of Ecology and Environment Science in Guangdong Higher Education, College of Life Science, South China Normal University, Guangzhou 510631, China;1. Kastamonu University, Faculty of Science and Art, Biology Department, 37100 Kuzeykent, Kastamonu, Turkey;2. Ookusa Animal Clinic, Matsue, Shimane 690-0032, Japan;1. The Key Laboratory of Biomedical Information Engineering, Ministry of Education, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an 710049, PR China;2. School of Life Sciences, SunYat-Sen (Zhongshan) University, Guangzhou 510275, PR China;3. Department of Life Sciences, Natural History Museum, London SW7 5BD, UK;1. Universität Salzburg, FB Organismische Biologie, Hellbrunnerstrasse 34, A-5020 Salzburg, Austria;2. Boise State University, Department of Biological Sciences, Boise, ID 83725-1515, USA;3. University of the West Indies, Electron Microscopy Unit, Kingston, Jamaica;4. Universität Kaiserslautern, FB Biologie, Erwin-Schrödingerstrasse 14, D-67663 Kaiserslautern, Germany |
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| 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 104 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. |
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| Keywords: | House mice Magnetic capture Quantitative PCR |
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