title: [Development of PCR assay for detection of Angiostrongylus cantonensis in Pomacea canaliculata].
abstract: OBJECTIVE
  To establish a PCR assay for detecting the third-stage larvae of Angiostrongylus cantonensis in Pomacea canaliculata.
  
  
  METHODS
  Polymerase chain reaction primers were designed by the software Lasergene, based on the specific cDNA of the third-stage larvae of A.cantonensis in Genbank. The total RNA was prepared from the third-stage larvae of A.cantonensis and of the snails by TRIzol one-step protocol. Amplification by RT-PCR was carried out following the kit protocol.
  
  
  RESULTS
  RT-PCR assay revealed a clear differentiation between infected and negative snails. When a mixture of the total RNA from the negative snails and the third-stage larvae of A.cantonensis was tested by the PCR assay, the detectable level was 128 pg RNA, a concentration close to one third-stage larva of A.cantonensis, minimum concentration that could be found by naked eyes. The minimum detected total RNA concentration of the third-stage larvae of A.cantonensis was 105 pg by PCR assay.
  
  
  CONCLUSIONS
  A PCR assay has been developed for detecting A.cantonensis larva in Pomacea canaliculata.
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai 200025, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xiao-nong
authors.initials: XN
authors.lastname: Zhou
authors.forename: He-xiang
authors.initials: HX
authors.lastname: Liu
authors.forename: Shan
authors.initials: S
authors.lastname: Lv
authors.forename: Li-sha
authors.initials: LS
authors.lastname: Li
authors.forename: Jin-xiang
authors.initials: JX
authors.lastname: Lin
authors.forename: You-song
authors.initials: YS
authors.lastname: Li
mesh_terms: D017185:Angiostrongylus cantonensis / Q000235:genetics* / Q000254:growth & development; D000818:Animals; D007814:Larva; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012908:Snails / Q000469:parasitology*; D017206:Strongylida Infections / Q000175:diagnosis / Q000469:parasitology*
chemical_list: D018510:RNA, Helminth