一种基于数字免疫原理的伤寒沙门菌高灵敏度检测方法  

Digital immune high sensitivity to detect the establishment of Salmonella typhi

作　　者：王冠 窦雪晨 张宁 邓橙 杜耀华 WANG Guan;DOU Xue-chen;ZHANG Ning;DENG Cheng;DU Yao-hua(Institute of Medical Support Technology,Academy of System Engineering of Academy of Military Science of Chinese PLA,Tianjin 300161,China)

机构地区：[1]军事科学院系统工程研究院卫勤保障技术研究所,天津300161

出　　处：《微生物学免疫学进展》2022年第1期35-40,共6页Progress In Microbiology and Immunology

摘　　要：目的设计一种基于数字化免疫显色原理的检测方法,实现伤寒沙门菌的高灵敏度快速检测。方法以伤寒沙门菌为检测对象,选取氨基磁珠对目标细菌进行富集,以偶联特异性抗体的量子点对目标细菌进行特异性荧光标记;设计制作反应腔室,在液相条件下完成对目标细菌的富集和特异性荧光标记;通过荧光显微成像和荧光特征自动提取,实现对目标细菌的“数字化”精确定量。结果利用单抗体夹心免疫复合物,建立了基于数字化免疫的伤寒沙门菌检测方法。氨基磁珠对伤寒沙门菌的富集效率>95%;特异性量子点可实现对伤寒沙门菌的标记,检测1000μL浓度为1×10^(3)cfu/mL的伤寒沙门菌时,磁珠最佳添加量为50μL,37℃温浴条件下最佳反应时间为30 min;伤寒沙门菌检测效率>90%,检测灵敏度达到2.635×10^(3)cfu/mL。结论基于数字免疫技术原理建立的免疫复合体系能够实现对于伤寒沙门菌的高灵敏度检测。Objective This article is designed with a method based on digital immunochrome techniques to achieve rapid detection of Salmonella typhi.Methods Salmonella typhi was used as the test object,amino magnetic beads were selected to enrich the target bacteria,and quantum dots with specific antibody were used to specifically label the target bacteria.The reaction was designed and fabricated to enrich the target bacteria in liquid phase.The"digital"accurate quantitative counting of target bacteria can be realized by fluorescence microscopic imaging and automatic extraction of fluorescence characteristics.Results A method for the detection of Salmonella typhi based on digital immunity was established by using the sandwich immune complex of single antibody.The enrichment efficiency of amino magnetic beads against Salmonella typhi was over 95%.Specific quantum dots can be used to label Salmonella typhi successfully.For the detection of 1000μL 10^(3)cfu/mL Salmonella typhi,the optimal dosage of magnetic beads was 50μL,and the optimal reaction time was 30 min under 37℃temperature bath.The detection efficiency of Salmonella typhi was>90%and the detection sensitivity reaches 2.635×10^(3)cfu/mL.Conclusion Immunohydrated system optimized based on digital immune technology can achieve high sensitivity testing of Salmonella typhi.

关 键 词：数字免疫 磁珠 量子点 微生物检测 高灵敏度 伤寒沙门菌 

分 类 号：Q93-331[生物学—微生物学]