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Introducing an optimized method for purifying Toxoplasma gondii genomic DNA from rat brain to increase diagnosis efficiency of Toxoplasma infection
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Setayesh Yasami-Khiabani   , Samira Choopani , Jalal Babaie  , Majid Golkar , Mohammad Sayyah * |
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Abstract: (825 Views) |
Background and Aim: Extraction of Toxoplasma gondii genomic DNA from rat brain is challenging due to low level of contamination and high fat content. Quantitative real-time polymerase chain reaction (RT-qPCR) is associated with false negative results despite detecting small amounts of the gene. By changing method of DNA extraction, we could decrease number of false negative results.
Methods: Brains of eleven male Wistar rats infected with Tehran strain of Toxoplasma parasite (with 8 weeks of parasite infection) and three uninfected rats were isolated. The whole brain was used to purify DNA instead of 20 mg brain in the conventional method. Amount of protein kinase was doubled and the incubation period increased to 6 times. Then DNA of the samples was measured by RT-qPCR.
Results and Conclusion: Only three of the eleven infected rats were infection positive by the conventional method, while nine mice were positive by the optimizing method.
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| Keywords: DNA extraction, RT-qPCR, REP-529 |
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Full-Text [PDF 1222 kb]
(340 Downloads)
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Type of Study: Original Research |
Subject:
Neuroscience
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