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目的建立可快速检测单核细胞增生李斯特菌(Listeria monocytogenes,LM)的上转发光免疫层析技术(up-converting phosphor technology based lateral flow assay,UPT-LF),即LM-UPT-LF。方法针对LM特异的p60蛋白制备单克隆抗体,并与上转发光纳米颗粒(up-converting phosphor nanoparticles,UCP-NPs)共价偶联作为结合物,采用双抗体夹心免疫层析模式建立LM-UPT-LF,并对其敏感性与特异性进行评价。结果建立了可快速检测LM的LM-UPT-LF平台,在样本<10 cfu、10~99 cfu、100~1000 cfu绝对LM污染量条件下可分别在培养20、18、16 h检出阳性。其他13种食源性致病菌在高浓度(109cfu/ml)时无非特异性交叉。结论所建立的LM-UPT-LF操作简便快速,具有良好的敏感性和特异性,适用于一线病原调查与检测。
OBJECTIVE To establish an up-converting phosphor technology based lateral flow assay (UPT-LF), LM-UPT-LF, for the rapid detection of Listeria monocytogenes (LM). Methods Monoclonal antibodies against LM-specific p60 protein were prepared and covalently coupled to up-converting phosphor nanoparticles (UCP-NPs) as a conjugate. Double-antibody sandwich immunochromatography was used to establish LM-UPT -LF, and its sensitivity and specificity were evaluated. Results The LM-UPT-LF platform for rapid detection of LM was established. Positive samples could be detected at 20, 18, 16 h after culture at 10 cfu, 10 ~ 99 cfu and 100 ~ 1000 cfu absolute LM. The other 13 food-borne pathogens had no non-specific crossover at high concentrations (109 cfu / ml). Conclusion The established LM-UPT-LF is simple and rapid in operation, has good sensitivity and specificity and is suitable for first-line pathogen investigation and detection.