Korean J Parasitol.  2010 Dec;48(4):297-301. 10.3347/kjp.2010.48.4.297.

Multiplex PCR Detection of Waterborne Intestinal Protozoa; Microsporidia, Cyclospora, and Cryptosporidium

• Lee SH
• Joung M
• Yoon S
• Choi K
• Park WY
• Yu JR

• KMID: 1104695
• DOI: http://doi.org/10.3347/kjp.2010.48.4.297

Abstract

Recently, emerging waterborne 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 101 to 102 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 parvum, 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.

Keyword

Cyclospora; Cryptosporidium; microsporidia; multiplex PCR; waterborne protozoa

MeSH Terms

Cryptosporidium/*isolation & purification
Cyclospora/*isolation & purification
DNA Primers/genetics
DNA Restriction Enzymes/metabolism
DNA, Protozoan/genetics/metabolism
Humans
Microsporidia/*isolation & purification
Parasitology/*methods
Polymerase Chain Reaction/*methods
Polymorphism, Restriction Fragment Length
Sensitivity and Specificity
Water/*parasitology