Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Multiplex PCR Detection of Waterborne Intestinal Protozoa: Microsporidia, Cyclospora, and Cryptosporidium

  • Lee, Seung-Hyun (Department of Microbiology, Konkuk University School of Medicine) ;
  • Joung, Mi-Gyo (Department of Environmental and Tropical Medicine, Konkuk University School of Medicine) ;
  • Yoon, Se-Joung (Department of Environmental and Tropical Medicine, Konkuk University School of Medicine) ;
  • Choi, Kyoung-Jin (Department of Environmental and Tropical Medicine, Konkuk University School of Medicine) ;
  • Park, Woo-Yoon (Department of Radiation Oncology, College of Medicine, Chungbuk National University) ;
  • Yu, Jae-Ran (Department of Environmental and Tropical Medicine, Konkuk University School of Medicine)
  • Received : 2010.09.27
  • Accepted : 2010.11.18
  • Published : 2010.12.31
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Abstract

Recently, emerging waterbome protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from $10^1$ to $10^2$ oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium paNum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

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References

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