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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
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
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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
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公开(公告)号：US08142996B2
公开(公告)日：2012-03-27
申请号：US12224388
申请日：2007-02-15
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。
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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。
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公开(公告)号：US06589735B1
公开(公告)日：2003-07-08
申请号：US09434557
申请日：1999-11-12
申请人： Markus Kostrzewa , Thomas Fröhlich , Thomas Wenzel
发明人： Markus Kostrzewa , Thomas Fröhlich , Thomas Wenzel
IPC分类号： C12Q168
CPC分类号： C12Q1/6858 , C12Q2563/167
摘要： The invention relates to a method of investigating by mass spectrometry the genetic material deoxyribonucleic acid (DNA) replicated by polymerase chain reaction (PCR), for the identification of known mutations and polymorphisms; it particularly relates to the analysis of single nucleotide polymorphisms (SNPs) by matrix assisted laser desorption and ionization (MALDI). The invention consists of using a set of nucleoside triphosphates for the selective PCR replication of the DNA in which one or more of the nucleoside triphosphates have been made much heavier by attaching a chemical group, but in such a way that the replication is not disturbed by the polymerase. In this way a single nucleotide polymorphism in DNA pieces with a length of about 40 to 50 bases can very easily be made visible by mass spectrometry without any further manipulation.
摘要翻译： 本发明涉及一种利用聚合酶链反应（PCR）复制的遗传物质脱氧核糖核酸（DNA）进行质谱分析，鉴定已知突变和多态性的方法; 它特别涉及通过基质辅助激光解吸和电离（MALDI）分析单核苷酸多态性（SNP）。本发明包括使用一组核苷三磷酸来进行DNA的选择性PCR复制，其中一个或多个核苷 通过连接化学基团使三磷酸盐变得更重，但是以这样的方式使复制不被聚合酶干扰。 以这种方式，长度为约40至50个碱基的DNA片段中的单核苷酸多态性可以非常容易地通过质谱法可见而无需任何进一步的操作。
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公开(公告)号：US11754572B2
公开(公告)日：2023-09-12
申请号：US13102212
申请日：2011-05-06
申请人： Markus Kostrzewa , Jochen Franzen
发明人： Markus Kostrzewa , Jochen Franzen
IPC分类号： C12Q1/04 , G01N33/68 , G01N33/569
CPC分类号： G01N33/6851 , G01N33/569
摘要： A method of detecting specified target microbes in different types of sample uses only one to two cultivation steps for the enrichment of the target microbes from the sample, preferably in selective culture media, combined with a mass spectrometric detection method that identifies the target microbes in mixtures with other microbes even if the target microbes account for only a small proportion of the mixture. The sample may be a food sample, a sample from bodies of water used for bathing, a soil sample, a swabbed sample, a stool sample, an impactor sample with collected aerosol particles, amongst many others. The detection method is several days faster than standard methods and less expensive.
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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
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。
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公开(公告)号：US20110272573A1
公开(公告)日：2011-11-10
申请号：US13103672
申请日：2011-05-09
申请人： Markus Kostrzewa , Jochen Franzen
发明人： Markus Kostrzewa , Jochen Franzen
IPC分类号： H01J49/40
CPC分类号： H01J49/0027 , H01J49/164 , H01J49/40
摘要： The invention relates to acquisition techniques for time-of-flight mass spectra with ionization of the analyte substances by matrix assisted laser desorption. Generally speaking, these acquisition techniques involve adding together a large number of individual time-of-flight spectra, each with restricted dynamic measuring range, to form a sum spectrum. The invention provides a method that improves, in particular, the reproducibility, the concentration accuracy and therefore the ability to quantify the mass spectra. Particular embodiments also increase the dynamic range of measurement. For this purpose, multiple series of mass spectra are acquired, whereby the energy density in the laser spot is increased in discrete steps. As a result, many ion signals saturate the detector and can therefore no longer be evaluated. However, it is possible to employ a technique in which the ion beam is increasingly defocused, or, secondly, to replace parts of the spectrum that are subject to saturation by intensity extrapolations from mass spectra acquired with lower energy density. In the first case, hundreds or thousands of individual mass spectra must be added together in order to increase the dynamic measuring range. In the second case, the finally acquired mass spectrum, with its replacements, forms a mass spectrum with a high dynamic measuring range, improved reproducibility and better concentration accuracy. The gradient of the increasing intensities of the ion signals, as a function of the energy density, supplies additional information about the proton affinity of the analyte ions. The concentration accuracy is enhanced because the increase in the number of proton donors in the ionization plasma leads to an increase in the ionization of those analyte substances that have a lower proton affinity.
摘要翻译： 本发明涉及通过基质辅助激光解吸附物质分析物质离子化的飞行时间质谱采集技术。 一般来说，这些采集技术包括将大量具有限制动态测量范围的单个飞行时间频谱相加在一起以形成和频谱。 本发明提供了一种特别改进重现性，浓度精度以及因此量化质谱的能力的方法。 具体实施例还增加了测量的动态范围。 为此，获得了多个质谱系列，从而在离散步骤中激光斑点的能量密度增加。 因此，许多离子信号使检测器饱和，因此不再能被评估。 然而，可以采用这样一种技术，其中离子束越来越散焦，或者其次，通过由具有较低能量密度获得的质谱的强度外推替代经受饱和的部分光谱。 在第一种情况下，为了增加动态测量范围，必须将数百或数千个单个质谱加在一起。 在第二种情况下，最终获得的质谱与其替代物形成具有高动态测量范围，改进的再现性和更好的浓度精度的质谱。 作为能量密度的函数的离子信号的增加强度的梯度提供关于分析物离子的质子亲和力的附加信息。 由于离子化等离子体中的质子供体的数量的增加导致具有较低质子亲和力的分析物质的电离增加，所以浓度精度得到提高。
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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。
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公开(公告)号：US20090305327A1
公开(公告)日：2009-12-10
申请号：US12224388
申请日：2007-02-15
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。
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