血清exosomes中蛋白、miRNAs、lncRNAs多指标联合用于血吸虫活虫感染诊断的可行性研究

Schistosomiasis is still an important public-health problem in China. Schistosomiasis lack effective diagnosis and curative evaluation methods. In our study, we intend to use mice serum exosomes as the research object. Serum exosomes are derived from control mice, schistosoma active infection mice, mice after curing at different time points. We detect differentially expressed proteins by 2D-nano-LC-MS/MS technology. Specific monoclonal antibodies of differentially expressed proteins are prepared. These antibodies are used to validate and further refine differentially expressed proteins in mice and human serum by double antibody sandwich ELISA. We also detect differentially expressed miRNAs and lncRNAs by high-throughput chip,establish different expression profiles of miRNAs and lncRNAs. Differentially expressed miRNAs and lncRNAs are validated and further refined in mice and human serum using qRT-PCR. Our research is intend to provide basic data and technical reserves for the effective diagnosis and epidemiological investigation of schistosomiasis.

日本血吸虫病仍是我国重要的公共卫生问题，缺乏有效诊断与疗效考核方法。本研究拟以正常对照、血吸虫活虫感染后、治愈后不同时间点小鼠血清exosomes为研究对象，采用2D-nano-LC-MS/MS技术筛选差异表达蛋白，建立差异蛋白表达谱，制备特异性单克隆抗体，运用双抗体夹心ELISA法在动物模型及人群血清中进行验证和进一步筛选；并用高通量miRNAs、lncRNAs芯片筛选差异表达的miRNAs、lncRNAs，建立差异表达谱，运用qRT-PCR法在动物模型及人群血清中进行验证和进一步筛选。为血吸虫病有效诊断和流行病学调查提供基础数据和技术储备。

本研究以正常对照、血吸虫活虫感染后、治愈后不同时间点小鼠血清exosomes为研究对象，采用2D-nano-LC-MS/MS技术筛选到3种差异表达蛋白，制备了特异性单克隆抗体，运用双抗体夹心ELISA法在动物模型及人群血清中进行验证和筛选，建立了一种血吸虫病诊断方法；并用高通量miRNAs、lncRNAs芯片筛选差异表达的miRNAs、lncRNAs，建立差异表达谱，运用qRT-PCR法在动物模型及人群血清中进行了验证和筛选。本研究可为血吸虫病有效诊断和流行病学调查提供基础数据和技术储备。