Objective To establish a simple and rapid detection technique for Oncomelania infected with Schistosoma japonicum(SJ), with high sensitivity and good specificity .Methods The gene fragment of SJ was amplified by PCR , and cloned into the T-vector to construct positive-reference.An isothermal nucleic acid amplification reaction system for detecting Oncomelania infected with SJ was set up , and its sensitivity was analyzed by detecting positive-reference diluted according to geometric proportion , and its specificity by detecting the genomic DNA of relative samples .Then, a corresponding means of purifying nucleic acid was designed to assemble a reagent detecting Oncomelania infected with SJ . This reagent was validated by detecting Oncomelania samples.Results The 213 bp amplified products were obtained and used to construct recombination T-vector for positive reference .An isothermal nucleic acid amplification reaction system was set up for detecting Oncomelania infected with SJ , and the amplification results could be simply determined by color change, with better sensitivity and specificity .The reagents for detecting Oncomelania infected with SJ were assembled , which could detect samples containing only 1% infected Oncomelania.Conclusion A visible detection method for Oncomelania infected with SJ is successfully established and validated .%目的 建立一种方便、快速且灵敏度高、特异性强的日本血吸虫( SJ )感染性钉螺检测方法. 方法 PCR扩增SJ基因片段,并将其克隆到T载体构建阳性参照物;建立SJ核酸恒温扩增反应体系,并通过检测经等比稀释的阳性参照物和相关样本基因组DNA分析其灵敏度和特异性;设计相应的核酸纯化方法,组装SJ感染性钉螺检测试剂,并通过检测钉螺样本对其进行验证. 结果 获得长度213 bp的PCR扩增产物,并用该扩增产物构建了重组T载体,即阳性参照物;建立了SJ核酸恒温扩增反应体系,该体系可根据颜色变化判定扩增结果,且灵敏度和特异性良好;组装了SJ感染性钉螺检测试剂,该试剂可检测出含1%阳性钉螺的钉螺样本. 结论 成功建立了一种可视化的SJ感染性钉螺检测方法,并通过应用对其进行了验证.
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