title: [Preparation of DNA from Cryptosporidium parvum oocysts for PCR detection].
abstract: OBJECTIVE
  To establish three methods of DNA extraction from Cryptosporidium parvum oocysts and test by PCR.
  
  
  METHODS
  After three freeze-thaw cycles, three kinds of templates were extracted from the oocysts by Chelex-100, phenol/chloroform or genomic DNA purification system kit, and used for PCR detection. According to the sequence of a C. parrum gene (L.16996), a pair of primers was designed and synthesized, and used for PCR. The sensitivity of the template by Chelex 100 method was also tested by PCR.
  
  
  RESULTS
  One 446 bp PCR product was observed by agarose gel electrophoresis for all three kinds of templates. The PCR sensitivity by Chelex-100 extracted DNA reached for detection of a specimen containing only 1/2 oocyst.
  
  
  CONCLUSIONS
  The three kinds of extraction can all be served as templates for PCR detection of C. parvum oocysts, while Chelex 100 method is simpler, quicker and more reliable for DNA extraction of the parasite.
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Center of Malaria, Schistosomiasis and Filariasis, Shanghai 200025, China.
authors.forename: Yu-juan
authors.initials: YJ
authors.lastname: Shen
authors.forename: Jian-ping
authors.initials: JP
authors.lastname: Cao
authors.forename: Wei-yuan
authors.initials: WY
authors.lastname: Lu
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Li
authors.forename: Hai-peng
authors.initials: HP
authors.lastname: Liu
authors.forename: Yu-xin
authors.initials: YX
authors.lastname: Xu
authors.forename: Xiao-nong
authors.initials: XN
authors.lastname: Zhou
authors.forename: Lin-hua
authors.initials: LH
authors.lastname: Tang
authors.forename: Shu-xian
authors.initials: SX
authors.lastname: Liu
mesh_terms: D000818:Animals; D002648:Child; D016785:Cryptosporidium parvum / Q000235:genetics*; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification*; D003967:Diarrhea / Q000469:parasitology; D005243:Feces / Q000469:parasitology; D006801:Humans; D033661:Oocysts / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan