›› 2006, Vol. 24 ›› Issue (5): 8-355.

• 实验研究 • Previous Articles     Next Articles

Development of PCR Assay for Detection of Angiostrongyluscantonensis in Pomacea canaliculata

ZHANG Yi1;ZHOU Xiao-nong1;LIU He-xiang1;Lv Shan1;
LI Li-sha2;LIN Jin-xiang2;LI You-song2   

  1. 1 National Institute of Parasitic Diseases，Chinese Center for Disease Control and Prevention，WHO Collaborating Center for Malaria，Schistosomiasis and Filariasis，Shanghai 200025，China；2 Fujian Provincial Center for Disease Control and Prevention，Fuzhou 350001，China
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-10-30 Published:2006-10-30

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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， mini-mum 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. Conclusion A PCR assay has been developed for detecting A.cantonensis larva in Pomacea canaliculata.

Key words: Pomacea canaliculata, PCR, cDNA, Angiostrongylus cantonensis, Larva