Detection of Schistosoma japonicum cercaria DNA by loop-mediated isothermal amplification / 中国血吸虫病防治杂志

ABSTRACT

Objective To establish a method for detecting cercaria of Schistosoma japonicum.Methods The loop-mediated isothermal amplification(LAMP)was used.The cercaria DNA of Schistosoma japonicum was extracted by using GeneReleaser.Four primers which recognized 6 distinct regions on the calcium-binding protein gene of cercaria of Schistosoma japonicum were designed and used for LAMP assay and Clonorchis sinensis was used as the negative control for evaluating the specificity and 20,10,5,1 cercariae of Schistosoma japonicum were amplified by LAMP for evaluating the sensitivity.The LAMP results were judged with the naked eyes and electrophoretic analysis.Results After LAMP reaction,the positive signal was observed with cercariae of Schistosoma japonicum.By contrast,no positive signal was observed for Clonorchis sinensis.The detection limit of LAMP assay was 1 cercaria of Schistosoma japonicum per reaction.Conclusion LAMP assay has usefulness for rapid detection of cercariae of Schistosoma japonicum.