悬浮芯片定量检测鼠疫菌的方法研究

Development of a suspension array method for quantitative detection of Y. pestis

目的利用编码微球悬浮芯片技术,探讨直接从"白色粉末"样品中检测鼠疫菌的可行性,为建立快速、敏感、特异、高通量、同时检测多种生物恐怖因子的技术平台奠定基础。方法用抗体包被编码微球作为反应载体,双抗体夹心法为反应模式,建立鼠疫菌F1抗原悬浮芯片检测方法,对"白色粉末"中的鼠疫耶尔森菌进行检测。通过盲样和标准实验室检测评估对现场样品的适用性。结果建立的定量检测鼠疫菌F1抗原的悬浮芯片方法具有较高的特异性和敏感性,并在0.154～4514ng/ml浓度范围内具有良好的动力学响应特性,比ELISA方法有较高的灵敏度和较宽动态检测范围。结论定量检测鼠疫菌的悬浮芯片方法能快速、敏感、特异地定量检测"白色粉末"中鼠疫菌,在早期识别、快速诊断和应对生物恐怖威胁、传染病暴发中具有广阔的应用前景。

Objective To establish a platform for rapid, sensitive, specific, high-throughput, and simultaneous detection of muhiple biothreat-associated agents, a suspension array-based immunoassay was developed for exploring the feasibility of directly simultaneous detection of Y. pestis in powder samples. Methods The immunoassay using coupled fluorescent beads were employed to detect Y. pestis as a model from powder sample and the feasibility for detection in field samples was demonstrated by both blind and standard laboratory trials. Results The newly developed suspension array appeared to be specific and sensitive, with the detection sensitivities of 0. 154ng/ml for Y. pestis F1 antigen, and the dynamic ranges of 0.154 -4514ng/ml, which were higher than those of corresponding conventional ELISA tests. Conclusion The suspension array could rapidly, sensitively, specifically and quantitatively detect pathogen from powder samples, which would be useful for early identification, rapid diagnosis, and response for the attack of bioterrorism and outbreak of infectious disease.