公开(公告)号：US20060095212A1

公开(公告)日：2006-05-04

申请号：US11249147

申请日：2005-10-12

申请人： Markus Kostrzewa ,  Stefan Klepel ,  Thomas Maier

发明人： Markus Kostrzewa ,  Stefan Klepel ,  Thomas Maier

IPC分类号： G06F19/00

CPC分类号： H01J49/0036 ,  Y10T436/143333

摘要： The invention generates mass scale comparability between mass spectra which are acquired in time-of-flight mass spectrometers, particularly with ionization by matrix-assisted laser desorption. Always slightly distorted mass scales of different mass spectra from the same type of sample can be aligned. The flight times of identical ions always differ slightly from one mass spectrum to the next due to non-reproducible processes in the ionization method. Thus the apparent mass values of ion signals of identical substances in different mass spectra do not match even if the flight times are converted into mass values with the identical calibration equation. After alignment of the mass scales, mass spectra can be reliably compared with respect to deviations in intensities of bio-makers, or be added together without deterioration in the mass resolution, and improved reference spectrum libraries can be created. Furthermore, the invention allows more reliable library searches to be carried out.

摘要翻译： 本发明在飞行时间质谱仪中获得的质谱之间产生质谱比较，特别是通过基质辅助激光解吸电离。 始终可以对齐来自相同类型样品的不同质谱的轻微变形质谱。 由于电离方法中不可重复的过程，相同离子的飞行时间总是与一个质谱图稍微不同。 因此，即使以相同的校准方程将飞行时间转换为质量值，不同质谱中相同物质的离子信号的表观质量也不匹配。 在对质量标尺进行比对后，质谱可以与生物制造者的强度偏差可靠地进行比较，或者加在一起，而不会降低质量分辨率，并且可以创建改进的参考光谱库。 此外，本发明允许执行更可靠的库搜索。

公开(公告)号：US20100248298A1

公开(公告)日：2010-09-30

申请号：US12695235

申请日：2010-01-28

申请人： Markus Kostrzewa ,  Thomas Maier ,  Stefan Klepel

发明人： Markus Kostrzewa ,  Thomas Maier ,  Stefan Klepel

IPC分类号： C12Q1/04

CPC分类号： G01N33/6848 ,  C12Q1/04 ,  G16B20/00 ,  Y02A50/58

摘要： Microorganisms are identified as present in a complex sample or mixed culture by acquiring a mass spectrum of the sample and comparing it to combination spectra, each of which is formed by combining at least two reference mass spectra of known microorganisms. Microorganisms corresponding to the reference spectra used to form the combination spectrum are identified as present in the sample if that combination spectrum exhibits a better match with the sample mass spectrum than any one of reference mass spectra used to form that combination spectrum. It is also possible to identify microorganisms by forming a difference spectrum by subtracting a reference mass spectrum from the sample mass spectrum and comparing the difference spectrum to the reference mass spectra.

摘要翻译： 通过获取样品的质谱并将其与组合光谱进行比较，将微生物鉴定为存在于复合物样品或混合培养物中，每个组合谱通过组合已知微生物的至少两个参考质谱而形成。 与用于形成组合光谱的参考光谱相对应的微生物被鉴定为存在于样品中，如果该组合光谱与样品质谱比与用于形成该组合光谱的参考质谱中的任何一个相比更好的匹配。 也可以通过从样品质谱中减去参考质谱并将差谱与参考质谱进行比较来形成差谱来鉴定微生物。

公开(公告)号：US07391017B2

公开(公告)日：2008-06-24

申请号：US11249147

申请日：2005-10-12

申请人： Markus Kostrzewa ,  Stefan Klepel ,  Thomas Maier

发明人： Markus Kostrzewa ,  Stefan Klepel ,  Thomas Maier

IPC分类号： B01D59/44 ,  H01J49/00

CPC分类号： H01J49/0036 ,  Y10T436/143333

摘要： The invention generates mass scale comparability between mass spectra which are acquired in time-of-flight mass spectrometers, particularly with ionization by matrix-assisted laser desorption. Always slightly distorted mass scales of different mass spectra from the same type of sample can be aligned. The flight times of identical ions always differ slightly from one mass spectrum to the next due to non-reproducible processes in the ionization method. Thus the apparent mass values of ion signals of identical substances in different mass spectra do not match even if the flight times are converted into mass values with the identical calibration equation. After alignment of the mass scales, mass spectra can be reliably compared with respect to deviations in intensities of bio-makers, or be added together without deterioration in the mass resolution, and improved reference spectrum libraries can be created. Furthermore, the invention allows more reliable library searches to be carried out.

摘要翻译： 本发明在飞行时间质谱仪中获得的质谱之间产生质谱比较，特别是通过基质辅助激光解吸电离。 始终可以对齐来自相同类型样品的不同质谱的轻微变形质谱。 由于电离方法中不可重复的过程，相同离子的飞行时间总是与一个质谱图稍微不同。 因此，即使以相同的校准方程将飞行时间转换为质量值，不同质谱中相同物质的离子信号的表观质量也不匹配。 在对质量标尺进行比对后，质谱可以与生物制造者的强度偏差可靠地进行比较，或者加在一起，而不会降低质量分辨率，并且可以创建改进的参考光谱库。 此外，本发明允许执行更可靠的库搜索。

公开(公告)号：US20110012016A1

公开(公告)日：2011-01-20

申请号：US12834504

申请日：2010-07-12

申请人： Thomas Maier ,  Markus Kostrzewa

发明人： Thomas Maier ,  Markus Kostrzewa

IPC分类号： B01D59/44

CPC分类号： G01N33/6848 ,  C12Q1/04 ,  G01N2560/00 ,  G06F19/24

摘要： 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.

摘要翻译： 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。

公开(公告)号：US10006076B2

公开(公告)日：2018-06-26

申请号：US14113354

申请日：2011-07-29

申请人： Jochen Franzen ,  Markus Kostrzewa ,  Thomas Maier ,  Karsten Michelmann ,  Wolfgang Pusch

发明人： Jochen Franzen ,  Markus Kostrzewa ,  Thomas Maier ,  Karsten Michelmann ,  Wolfgang Pusch

IPC分类号： C12Q1/04 ,  G01N33/68 ,  G01N21/77

CPC分类号： C12Q1/04 ,  G01N21/77 ,  G01N33/6851 ,  G01N2021/7786 ,  G01N2800/26

摘要： 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.

公开(公告)号：US08237107B2

公开(公告)日：2012-08-07

申请号：US12834504

申请日：2010-07-12

申请人： Thomas Maier ,  Markus Kostrzewa

发明人： Thomas Maier ,  Markus Kostrzewa

IPC分类号： B01D59/44 ,  H01J49/00

CPC分类号： G01N33/6848 ,  C12Q1/04 ,  G01N2560/00 ,  G06F19/24

摘要： 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.

摘要翻译： 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。

公开(公告)号：US10144946B2

公开(公告)日：2018-12-04

申请号：US13102085

申请日：2011-05-06

申请人： Katrin Sparbier ,  Markus Kostrzewa

发明人： Katrin Sparbier ,  Ulrich Weller ,  Markus Kostrzewa

IPC分类号： C12Q1/04 ,  G01N33/68

摘要： The invention relates to the detection of specified, flagellated bacteria, particularly Salmonella, in food and stool. A single culturing period of about 12 to 24 hours in a liquid nutrient medium without agitation is combined with a position-selective sampling of the flagellated microbes from the liquid of the culture, after which a mass spectrometric detection method is used which recognizes the target bacteria in mixtures. A second culture step is only necessary in exceptional cases. A species-selective or genus-selective culture medium is advantageous. Positional selection becomes possible because these bacteria use their flagella to counteract sedimentation by chemotaxis, and they collect near the surface. This provides a low-cost detection method that is several days faster than conventional methods

公开(公告)号：US20120107864A1

公开(公告)日：2012-05-03

申请号：US13102085

申请日：2011-05-06

申请人： Katrin Sparbier ,  Markus Kostrzewa

发明人： Katrin Sparbier ,  Markus Kostrzewa

IPC分类号： C12Q1/04

CPC分类号： C12Q1/04 ,  G01N2560/00 ,  Y02A50/451

摘要： The invention relates to the detection of specified, flagellated bacteria, particularly Salmonella, in food and stool. A single culturing period of about 12 to 24 hours in a liquid nutrient medium without agitation is combined with a position-selective sampling of the flagellated microbes from the liquid of the culture, after which a mass spectrometric detection method is used which recognizes the target bacteria in mixtures. A second culture step is only necessary in exceptional cases. A species-selective or genus-selective culture medium is advantageous. Positional selection becomes possible because these bacteria use their flagella to counteract sedimentation by chemotaxis, and they collect near the surface. This provides a low-cost detection method that is several days faster than conventional methods

摘要翻译： 本发明涉及在食物和粪便中检测特定的鞭毛细菌，特别是沙门氏菌。 将不搅拌的液体营养培养基中的约12至24小时的单次培养期与来自培养液的鞭毛微生物进行位置选择性取样组合，之后使用识别目标细菌的质谱检测方法 在混合物中。 第二个文化步骤仅在特殊情况下才有必要。 物种选择性或属选择性培养基是有利的。 位置选择成为可能，因为这些细菌使用其鞭毛来抵抗趋化性的沉降，并且它们在表面附近收集。 这提供了比常规方法快几天的低成本检测方法

公开(公告)号：US08142996B2

公开(公告)日：2012-03-27

申请号：US12224388

申请日：2007-02-15

申请人： Jochen Franzen ,  Karsten Michelmann ,  Markus Kostrzewa

发明人： Jochen Franzen ,  Karsten Michelmann ,  Markus Kostrzewa

IPC分类号： C12Q1/00 ,  G01N33/49

CPC分类号： G01N33/6848 ,  C12Q1/37 ,  C12Q1/56 ,  G01N2333/75 ,  G01N2333/976

摘要： The invention relates to the determination of the nature and strength of enzymatic activity in blood using mass spectrometric measurement of a profile of the reaction products. The determination of the enzymatic activity can be used for medical diagnostics, for example, and also to check the effectiveness of medication. The invention provides a method whereby adding probe substances usually not present in blood offers standardized substrates for measuring the enzymatic activity. The probe substances may be added to whole blood, plasma, or serum. The mass spectrometric measurement of the reaction products, after their reversible immobilization on actively binding surfaces of solids, for example, can deliver biomarker patterns of the reaction products which may be indicators for metabolic anomalies or diseases, since these are often accompanied by the formation or activation of characteristic enzymes.

摘要翻译： 本发明涉及使用反应产物分布的质谱测量来测定血液中酶活性的性质和强度。 酶活性的测定可以用于医学诊断，例如，以及检查药物的有效性。 本发明提供了通过添加通常不存在于血液中的探针物质来提供用于测量酶活性的标准化底物的方法。 探针物质可以添加到全血，血浆或血清中。 反应产物在其可固定地固定在固体的活性结合表面上之后的质谱测量可以提供反应产物的生物标志物模式，其可以是代谢异常或疾病的指标，因为它们通常伴随着形成或 特征酶的活化。

公开(公告)号：US20110202282A1

公开(公告)日：2011-08-18

申请号：US13005863

申请日：2011-01-13

申请人： Markus Kostrzewa

发明人： Markus Kostrzewa

IPC分类号： G06F19/00

CPC分类号： G06F19/28 ,  C12Q1/04 ,  G01N33/6848 ,  H01J49/0036

摘要： Microbes in a sample are identified by calculating similarities between a mass spectrum of the sample and reference mass spectra in a spectral library. The spectral library is divided into a hierarchy of sub-libraries where each sub-library contains reference mass spectra of microbes which are statistically the most prevalent in the samples, but are not included in other sub-libraries and all additional reference mass spectra in the library that have substantial similarity to the reference mass spectra of these microbes. Only if the search in a sub-library does not provide a hit with sufficient certainty of identification, is the search carried out in sub-libraries of higher stages.

摘要翻译： 通过计算样品的质谱和光谱库中的参考质谱之间的相似性来鉴定样品中的微生物。 光谱库被划分成子库的层次结构，其中每个子库包含在样品中统计学上最普遍的微生物的参考质谱图，但不包括在其他子文库中，并且所有其他参考质谱图 文库与这些微生物的参考质谱图具有很大的相似性。 只有在子图书馆中进行搜索并没有提供足够的识别确定性的情况下，才能在更高阶段的子图书馆中进行搜索。