title: Multiplex PCR detection of waterborne intestinal protozoa: microsporidia, Cyclospora, and Cryptosporidium.
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 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 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.
authors.affiliation: Department of Microbiology, Konkuk University School of Medicine, Seoul 143-701, Korea.
authors.forename: Seung-Hyun
authors.initials: SH
authors.lastname: Lee
authors.forename: Migyo
authors.initials: M
authors.lastname: Joung
authors.forename: Sejoung
authors.initials: S
authors.lastname: Yoon
authors.forename: Kyoungjin
authors.initials: K
authors.lastname: Choi
authors.forename: Woo-Yoon
authors.initials: WY
authors.lastname: Park
authors.forename: Jae-Ran
authors.initials: JR
authors.lastname: Yu
mesh_terms: D003458:Cryptosporidium / Q000302:isolation & purification*; D021744:Cyclospora / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004262:DNA Restriction Enzymes / Q000378:metabolism; D016054:DNA, Protozoan / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016814:Microsporidia / Q000302:isolation & purification*; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D014867:Water / Q000469:parasitology*
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan; D014867:Water; D004262:DNA Restriction Enzymes