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    • 59. 发明授权
    • ATP-bioluminescence immunoassay
    • ATP生物发光免疫测定
    • US08518658B1
    • 2013-08-27
    • US12767946
    • 2010-04-27
    • Daniel V. LimDawn M. Hunter
    • Daniel V. LimDawn M. Hunter
    • C12Q1/66
    • G01N33/56916C12Q1/04C12Q1/10C12Q1/14G01N33/56938G01N33/56944Y02A50/451
    • Disclosed is a method and associated device for the rapid identification of viable bacterial contaminants in food products. The method detects viable microbes by using a combined ATP-bioluminescence immunoassay. Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium were selected as target organisms in various matrices including ground beef homogenate, apple juice, milk, and phosphate-buffered saline. Specific antibodies were immobilized on the surface of well plates in which the sample matrices were incubated. The plates were washed, and the wells were incubated with BacTiter-Glo reagent in Mueller-Hinton II broth. Bioluminescent output was measured with a luminometer and signal-to-noise ratios were calculated. The LOD was not affected by the presence of non-target cells. A strong linear correlation was observed between the number of cells and luminescent output over 4 orders of magnitude. This method provides a means of simultaneously detecting and identifying viable pathogens in complex matrices.
    • 公开了用于快速鉴定食品中的活细菌污染物的方法和相关装置。 该方法通过使用组合的ATP-生物发光免疫测定来检测活微生物。 选择大肠杆菌O157:H7和沙门氏菌血清型鼠伤寒沙门氏菌作为各种基质的靶生物,包括研磨牛肉匀浆,苹果汁,牛奶和磷酸盐缓冲盐水。 将特异性抗体固定在其中样品基质孵育的孔板的表面上。 洗涤板,并将孔与Mueller-Hinton II肉汤中的BacTiter-Glo试剂一起温育。 用发光计测量生物发光输出,并计算信噪比。 LOD不受非靶细胞的存在的影响。 在4个数量级的细胞数和发光量之间观察到强烈的线性相关性。 该方法提供了一种在复杂基质中同时检测和鉴定活的病原体的方法。