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    • 4. 发明申请
    • MASS SPECTROMETRIC DIAGNOSIS OF SEPSIS WITHOUT BLOOD CULTURE
    • 没有血液文化的食物的大量光谱诊断
    • US20160251694A1
    • 2016-09-01
    • US14113354
    • 2011-07-29
    • Jochen FranzenMark KostrzewaThomas MaierKarsten MichelmannWolfgang Pusch
    • Jochen FranzenMark KostrzewaThomas MaierKarsten MichelmannWolfgang Pusch
    • C12Q1/04G01N21/77
    • C12Q1/04G01N21/77G01N33/6851G01N2021/7786G01N2800/26
    • The invention relates to methods and instruments for the rapid detection and rapid mass spectrometric identification of microbial infective agents in blood or other body fluids. The invention recognizes that blood is not a good environment for the cultivation of microbes and provides a method which (a) largely destroys or dissolves the human particles in body fluids, such as erythrocytes and leukocytes in blood, without impairing the ability of the microbes to reproduce, (b) separates the microbial pathogens from the fluid, (c) cultivates them in a nutrient broth which contains none of the antimicrobial components of the body fluids, (d) separates them from the nutrient broth, and (e) identifies the microbes by a mass spectrum of the microbial proteins. The dissolution of the human particles also releases the microbes nesting in macrophages. The cultivation in an optically clear nutrient broth with optimum composition not only accelerates the propagation of the microbes compared to all other cultivation methods, but also makes it possible to continuously measure their quantitative growth starting from a low microbe density. This firstly allows the mass spectrometric identification to be carried out at the earliest possible time, secondly provides a positive detection of microbes far ahead of their identification, which can be lifesaving for the patient; and thirdly makes it possible to start the determination of resistances early.
    • 本发明涉及快速检测和快速质谱鉴定血液或其他体液中微生物感染因子的方法和仪器。 本发明认识到,血液不是培养微生物的良好环境,并且提供了(a)在体内流体中大量地破坏或溶解血液中的红细胞和白细胞的方法,而不会损害微生物的能力 (b)将微生物病原体与流体分离,(c)将其培养在不含体液抗微生物成分的营养肉汤中,(d)将其与营养液分离,以及(e) 微生物通过微生物蛋白质的质谱。 人类颗粒的溶解也释放出嵌入巨噬细胞的微生物。 在具有最佳组成的光学透明营养肉汤中培养不仅加速微生物与所有其他培养方法相比的繁殖,而且可以从低微生物密度开始连续测量其定量生长。 这首先允许在早期进行质谱鉴定,其次提供远远超过其鉴定的微生物的阳性检测,这可以为患者挽救; 第三,可以提前开始确定电阻。
    • 7. 发明申请
    • MASS SPECTROMETRIC DIAGNOSIS OF SEPTICEMIA
    • 大分子诊断分析
    • US20120115182A1
    • 2012-05-10
    • US13322363
    • 2010-07-14
    • Thomas Maier
    • Thomas Maier
    • C12Q1/04
    • C12Q1/24C12Q1/04G01N33/6851G01N2560/00G01N2570/00G01N2800/26
    • The invention mainly relates to the mass spectrometric identification of pathogens in blood cultures from blood-stream infections (septicemia). The invention provides a method with which microbial pathogens can be separated in purified form from blood after a relatively brief cultivation in a blood culture flask, without any interfering human proteins or any residual fractions of blood particles such as erythrocytes and leukocytes, and can be directly identified by mass spectrometric measurement of their protein profiles. The method is based on the use of relatively strong tensides to destroy the blood particles by dissolving the weak cell membranes and most of the internal structures of the blood particles; in spite of the fact that tensides are regarded as strong ionization inhibitors in MALDI and other ionization processes required for mass spectrometric measurements. This method allows unknown pathogens to be obtained in their pure form by centrifuging or filtration and to be identified on the taxonomic level of species or subspecies. Problems with DNA from high levels of leukocytes can be resolved by special measures. After sufficient cultivation, the identification in a mass spectrometric laboratory takes only half an hour.
    • 本发明主要涉及血流感染(败血病)血液培养物中病原体的质谱鉴定。 本发明提供了一种在血培养瓶中比较短暂的培养后,将微生物病原体与血液分离的方法,其中没有任何干扰的人类蛋白质或血液颗粒如红细胞和白细胞的任何残留部分,并且可直接 通过其蛋白质谱的质谱测定来鉴定。 该方法基于使用相对较强的表面活性剂通过溶解弱细胞膜和血液颗粒的大部分内部结构来破坏血液颗粒; 尽管在MALDI和质谱测量所需的其他电离过程中,表面活性剂被认为是强电离抑制剂。 该方法允许通过离心或过滤以其纯形式获得未知的病原体,并根据物种或亚种的分类级别进行鉴定。 通过特殊措施可以解决高水平白细胞DNA的问题。 在充分培养后,质谱实验室的鉴定只需半小时。
    • 10. 发明申请
    • SPECTROPHOTOMETRIC IDENTIFICATION OF MICROBE SUBSPECIES
    • 微量元素的分光光度鉴定
    • US20110012016A1
    • 2011-01-20
    • US12834504
    • 2010-07-12
    • Thomas MaierMarkus Kostrzewa
    • Thomas MaierMarkus Kostrzewa
    • B01D59/44
    • G01N33/6848C12Q1/04G01N2560/00G06F19/24
    • A dual-stage method is provided for identifying a microbe by, for example, its species or its subspecies. The method includes measuring a mass spectrum of the microbe using a mass spectrometer, calculating indicators for similarities between reference mass spectra in a library and the measured mass spectrum, selecting a group of reference mass spectra similar to the measured mass spectrum, determining a distinguishing weight for each mass signal of the reference mass spectra in the group, where the distinguishing weights emphasize differences between the reference mass spectra in the group, and calculating indicators for similarities between the reference mass spectra in the group and the measured mass spectrum as a function of the distinguishing weights.
    • 提供了用于通过例如其物种或其亚种鉴定微生物的双阶段方法。 该方法包括使用质谱仪测量微生物的质谱,计算文库中参考质谱与测量质谱之间的相似性的指标,选择与测得的质谱相似的一组参考质谱,确定鉴别重量 对于组中参考质谱的每个质量信号,其中区分权重强调组中参考质谱之间的差异,并且计算组中参考质谱与测量质谱之间的相似性的指标,作为 区别权重。