[Development of a PCR assay for detecting Schistosoma japonicum-infected Oncomelania hupensis].
OBJECTIVE
To establish a sensitive and specific PCR assay for detecting Schistosoma japonicum-infected Oncomelania hupensis.


METHODS
Based on 18S-rRNA gene of S. japonicum, a PCR assay for detecting Oncomelania snails infected with S. japonicum was established. The PCR product was sequenced, and the sensitivity, cross-reaction and mass detection experiments of PCR assay were performed.


RESULTS
The location of PCR product for detecting Oncomelania snails infected with S. japonicum was similar to the target DNA, with a length of 469 bp and the same sequence as the target DNA. It was registered in GenBank (Accession No. DQ442999). There was no PCR product for detecting uninfected snail. Experiments showed that the minimum DNA concentration of S. japoncium miracidium to be detected was 40 pg/Rpl. DNA from snail infected with single-tail cercaria could not be detected. The maximum dilution concentration of infected snail DNA pooled with uninfected snail DNA that could be detected was 1:640.


CONCLUSIONS
The PCR assay for detecting S. japonicum-infected Oncomelania snails shows high sensitivity, specificity and effect of mass detection.
Chen
Jun-hu
JH
Institute of Parasitic Diseases, Zhejiang Academy of Medical Sciences, WHO Collaborating Center for Research on Helminthiasis, Hangzhou 310013, China.
Wen
Li-yong
LY
Zhang
Xu-zhao
XZ
Zhang
Jian-feng
JF
Yu
Li-ling
LL
Hong
Lin-di
LD
D000818:Animals; D001483:Base Sequence; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S; D012549:Schistosoma japonicum / Q000235:genetics*; D012908:Snails / Q000469:parasitology*
D012337:RNA, Ribosomal, 18S