Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
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A highly sensitive molecular diagnosis method for detecting Toxoplasma gondii tachyzoite: a PCR/dot blot hybridization

  • Hong, Sun-Hwa (Center for Animal Resources Development, Wonkwang University) ;
  • Lee, Yun-Seong (Center for Animal Resources Development, Wonkwang University) ;
  • Kim, Young-Ho (Department of Biochemistry, School of Medicine, Wonkwang University) ;
  • Kim, Ok-Jin (Center for Animal Resources Development, Wonkwang University)
  • Received : 2013.08.13
  • Accepted : 2014.01.15
  • Published : 2014.03.30
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Abstract

This study aimed at finding a fast, sensitive, and efficient protocol for molecular identification of intracellular protozoa Toxoplasma (T.) gondii. For molecular detection of T. gondii, we developed a polymerase chain reaction coupled with dot blot hybridization assay (PCR/DBH). For DBH analysis, the amplified DNA of T. gondii tachyzoite was labeled by incorporation of digoxigenin. The DBH assay alone was capable of detecting down to $1{\times}10^4$ pg of T. gondii genomic DNA. The PCR alone was capable of detecting down to $1{\times}10^3$ pg of T. gondii genomic DNA, whereas the PCR/DBH assay was capable of detecting down to $1{\times}10^2$ pg of T. gondii genomic DNA, indicating that sensitivity of the PCR/DBH method was approximately 10 to 100 times higher than PCR or DBH alone. Our PCR/DBH assay will be useful for confirming the presence of T. gondii on the samples and differentiating T. gondii infection from other intracellular protozoa infections.

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References

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