日本血吸虫感染分子诊断技术的研究

Detection of Schistosoma Japonicum Infection by Nested-PCR

目的建立检测日本血吸虫感染的巢式PCR技术。方法设计2对巢式引物特异性扩增日本血吸虫高拷贝的Sjα1基因片段,分析反应的敏感性和特异性,并对感染小鼠血清、全血样本以及实验和现场钉螺样本进行检测。结果建立的巢式PCR方法特异性扩增日本血吸虫420bp的Sjα1片段,和曼氏血吸虫没有交叉,基因组DNA作为模板时最低检测量为0.1fg。小鼠感染日本血吸虫后2周的血清样本即能检测出特异性DNA,建立的方法能同时检测血清和全血标本。钉螺实验感染4hr后能检测到日本血吸虫DNA,现场采集钉螺的检测结果显示比传统的镜检方法敏感性高。结论建立的巢式PCR检测日本血吸虫感染具有较高的敏感性和特异性,为疾病诊断和媒介调查提供了新的分子生物学检测技术。

Objective To develop a nested-PCR for specific detection of Schistosoma japonicum.Methods Two pairs of nested-primers were designed targeted the Schistosoma japonicum with highly repeated Sjα1 gene sequence.The sensitivity and specificity of the nested-PCR were analysed. The samples from experimentally infected mice and Oncomelania snails were tested by the nested-PCR. Results A 420 bp DNA fragment was specifically amplified by the nested-PCR. The detection limit was 0.1fg when genomic Schistosoma japonicum...