【摘要】 目的采用纳米免疫磁珠分离副溶血性弧菌,建立副溶血性弧菌环介导等温扩增检测方法。方法采用副溶血性弧菌单克隆抗体,制备纳米免疫磁珠,特异性吸附副溶血性弧菌,结合环介导等温扩增技术,建立副溶血性弧菌快速检测方法。结果副溶血性弧菌纳米免疫磁珠在菌体浓度为103cfu/ml水平时,对副溶血性弧菌的捕获率达到74%。免疫磁分离结合环介导等温扩增技术,在纯培养、无需增菌情况下,检测灵敏度达到140 cfu/ml增菌液;通过对134株副溶血性弧菌和74株非目标菌的测试,环介导等温扩增技术具有良好的特异性;食品基质添加试验中,在增菌时间缩短至8 h的条件下,其检测限为2 cfu/25 g样品。结论副溶血性弧菌免疫纳米磁珠结合环介导等温扩增技术,有效缩短了增菌时间,适用于副溶血性弧菌的快速检测。更多还原

【Abstract】 Objective Anoval method of nano-immunomagnetic separation(Nano-IMS) plus loop-mediated isothermal amplification(LAMP) was established for the rapid detection of Vibrio parahemolyticus.Methods The Nano-Immunomagnetic Beads(Nano-IMB) were created by using the monoclonal antibody of V.parahemolyticus and named Nano-IMB-Vp,which was highly specific to V.parahemolyticus.The method of Nano-IMS-LAMP was established for the rapid detection of V.parahemolyticus from seefoods.Results The capture ratio of Nano-IMB-Vp was reached 74% at the level of 103 cfu/ml.In pure culture,the sensitivity of Nano-IMS-LAMP was reached 140 cfu/ml culture medium.The specific of this LAMP method was tested by using 134 targets and 74 non-targets bacteria.The results showed that the LAMP method was highly specific to V.parahemolyticus.Nocross-reaction was founded.The detection limit of Nano-IMS-LAMP reached 2 cfu/25 g in artficial samples when the period of time for enrichment was shorted to 8 h.Conclusion The method of Nano-IMS-LAMP could shorten the period of time for enrichment effectively,which can apply to the rapid detection of V.parahemolyticus.更多还原