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The Development of Lyophilized Loop-mediated Isothermal Amplification Reagents for the Detection of Coxiella burnetii
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[Detection of Toxoplasma gondii DNA by loop-mediated isothermal amplification]
Objective:
To detect Toxoplasma gondii DNA by loop-mediated isothermal amplification (LAMP).
Methods:
DNA was extracted by phenol-chloroform extraction from T. gondii tachyzoites. Four primers which recognized 6 distinct regions on the B1 gene of T. gondii were designed and used for LAMP assay. To evaluate the specificity of the method, Plasmodium vivax, P. falciparum, Pneumocystis carinii, Schistosoma japonicum, and mouse leucocytes were used gs controls. The parasite extract (T. gondii) was 10-fold serially diluted for evaluating the sensitivity of LAMP, and was amplified by LAMP. LAMP results were read with naked eye and analyzed by electrophoresis.
Results:
After LAMP reaction, positive amplification was observed with T. gondii, but no positive signal was noted for the negative controls in the study. The sensitivity of LAMP assay reached up to 2-3 T. gondii tachyzoites/ml per reaction.
Conclusion:
LAMP assay shows proper specificity and sensitivity for the detection of T. gondii.
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