基因芯片检测日本血吸虫及其现场应用

Development and Preliminary Application of the Genechip for Detection of Schistosoma japonicum

目的:建立敏感、特异的检测日本血吸虫的基因芯片并对其应用价值进行评价。方法:根据日本血吸虫高度保守的编码免疫原性毛蚴抗原5D基因,筛选、设计聚合酶链反应(PCR)的引物和基因探针,制成日本血吸虫基因芯片。采用基因芯片对江西、湖南和安徽三省现场采集的钉螺进行检测。检测时抽提待测钉螺的DNA,进行PCR扩增及不对称PCR荧光标记,然后与检测芯片杂交,最后进行扫描及分析。结果:建立的基因芯片能特异、敏感地检测出感染性钉螺,江西、湖南感染性钉螺的检出率为100％,安徽省品系的感染性钉螺检出率略低。结论:成功地建立了检测日本血吸虫的基因芯片,具有较高的实际应用价值。

Objective: To develop an accurate and highly specific assay of genechip for detection of Schistosoma japonicum (Sj. ) and evaluate the application value of the genechip. Methods; A conservative and specific gene sequence of an immunogenic miracidial 5D antigen of Sj. was used as probe and synthesized to be spotted on the microarray. After being extracted from sample Oncomelanian snail, a DNA fragment was amplified and labeled with fluorescence Cy3 -dUTP by dissymmetrical PCR and hybridized with the genechip, and then the genechip was scanned and analyzed. Results: The genechip showed significantly higher positive detection rate . The positive rates of areas from Jiangxi, Hunan in these positive subjects were all 100% (30/30, 20/20). The positive rate of Anhui was 85. 7%(18/21). Conclusion; The genechip can detect Oncomelanian containing Sj. with an adequate sensitivity and specificity, and it may be used for large - scale investigation.