摘要
目的采用Luminex液相悬浮芯片检测方法,建立一管同时检测19种葡萄球菌肠毒素和蜡样芽胞杆菌肠毒素的快速筛查方法。方法根据GenBank公布的葡萄球菌肠毒素entA-entR基因、蜡样芽胞杆菌cesB,nheB基因,设计特异性引物和探针,优化反应体系和条件,建立19种肠毒素的Luminex液相悬浮芯片检测法。结果 Luminex液相悬浮芯片体系检测19种肠毒素互不干扰,经验证80株金葡菌和蜡样芽胞杆菌菌株,均出现特异性荧光中值信号。Lu minex液相悬浮芯片技术从样品处理到检测结果需3.5h左右。结论 Luminex液相悬浮芯片筛查方法可应用于食物中毒等应急检测的快速和高通量筛查。
Objective To develop a Luminex liquid chip assay for the simultaneous detection of 19 enterotoxins in a single-tube reaction.Methods Specific primers and probes were designed and synthesized for entA-entR gene of Staphyloccocus,cesB and nheB gene of Bacillus cereus based on the target gene sequences from GenBank..Then the Luminex liquid chip assay for the detection of 19 enterotoxins were developed.Results The developed assay were tested against 80 strains and no cross-reaction and false signals were observed using Luminex liquid chip assay.Conclusion The Luminex liquid chip assay could be applied to the identification of 19 enterotoxins and the assay would be finished within 3.5 hours including samples DNA preparation and detection.The method could be applied to the rapid and simultaneous detection of enerotoxins in food poisoning.
出处
《中国热带医学》
CAS
2012年第11期1307-1309,1316,共4页
China Tropical Medicine
基金
国家自然科学基金项目(No.81071433)
广东省医学科学技术研究基金(No.B2012323)
深圳市科技计划项目(No.200902085)
