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目的:寻找一种有效的日本血吸虫病诊断及疗效考核方法。方法:以血吸虫的成虫、虫卵、尾蚴DNA为模板,利用聚合酶链反应(PCR)对日本血吸虫编码免疫原性毛蚴抗原的5D基因进行扩增,检测其敏感性与特异性。结果:用PCR检测日本血吸虫的成虫、虫卵、尾蚴DNA,均出现明显特异性条带。通过引物1、引物2单扩增,即能检测到单个虫卵、单个尾蚴或针尖大小的成虫组织。采用Nest-PCR和非同位素探针能将敏感性进一步提高。常见的肠道细菌及人基因组DNA无交叉阳性出现。结论:PCR方法检测日本血吸虫基因的敏感性与特异性均满意。
Objective: To find an effective method for diagnosis and efficacy evaluation of schistosomiasis japonica. Methods: The 5D gene of Schistosoma japonicum encoding the immunogenic miRNA antigen of Schistosoma japonicum was amplified by polymerase chain reaction (PCR) using the DNA of adult schistosomiasis, eggs and cercariae as templates. The sensitivity and specificity of 5D gene were determined. Results: PCR was used to detect the adult worm, eggs and cercaria DNA of Schistosoma japonicum, all showing obvious specific bands. By primer 1, primer 2 single amplification, that can detect a single egg, a single cercaria or needle-sized adult tissue. Sensitivity can be further enhanced with Nest-PCR and non-isotopic probes. Common intestinal bacteria and human genomic DNA without positive cross appears. Conclusion: The sensitivity and specificity of PCR to detect Schistosoma japonicum gene are satisfactory.