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1.发明申请公开(公告)号：US20130095511A1
公开(公告)日：2013-04-18
申请号：US13582372
申请日：2011-06-10
IPC分类号： C12Q1/18
CPC分类号： C12Q1/18 , C12Q1/34 , G01N33/6851 , G01N2333/986 , G01N2800/26 , G01N2800/44
摘要： The invention relates to the determination of resistances of microorganisms which produce β-lactamases, in particular “extended spectrum β-lactamases” (ESBL). The invention provides a method whereby the microbial resistance can be measured very simply and quickly by means of the catalytic effect of the microbially produced β-lactamases on β-lactam antibiotics, which consists in a hydrolytic cleavage of the β-lactam ring. The method determines the resistance of the bacteria a few hours after a suitable substrate, either a β-lactam antibiotic or a customized β-lactam derivative, has been added to a suspension of the microbes, by direct mass spectrometric measurement of the substrate breakdown caused by the β-lactamases.
摘要翻译： 本发明涉及测定产生β-内酰胺酶的微生物的抗性，特别是“延长光谱β-内酰胺酶”（ESBL）。 本发明提供了一种通过微生物产生的β-内酰胺酶对β-内酰胺抗生素的催化作用非常简单和快速地测量微生物抗性的方法，其中β-内酰胺抗生素包括β-内酰胺环的水解裂解。 该方法通过直接质谱测量引起的底物破坏，在合适的底物（β-内酰胺抗生素或定制的β-内酰胺衍生物）已添加到微生物的悬浮液中后几小时确定细菌的抗性 由β-内酰胺酶。
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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：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。
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8.发明授权公开(公告)号：US07160680B2
公开(公告)日：2007-01-09
申请号：US10079043
申请日：2002-02-20
CPC分类号： C12Q1/6858 , C12Q2565/627 , C12Q2523/319
摘要： The invention relates to a method of a mass-spectrometric analysis of known mutation sites in the genome, such as single nucleotide polymorphisms (SNPs), using the method of restricted primer extension. The invention consists of the use of primers with a photocleavable linker. The linker creates a gap in a DNA strand which is almost the same size as a natural DNA building block (nucleoside). The linker forms a bridge over a base pair without inhibiting hybridization or enzymatic extension. However, the linker allows the primers to be cleaved after extension in order to obtain short DNA fragments which can be more easily detected on the mass spectrometer.
摘要翻译： 本发明涉及使用限制性引物延伸的方法对基因组中已知突变位点进行质谱分析的方法，例如单核苷酸多态性（SNP）。 本发明包括使用具有光可透洗接头的引物。 接头在DNA链中产生与天然DNA构建块（核苷）几乎相同大小的间隙。 连接体在碱基对上形成桥，而不抑制杂交或酶促延伸。 然而，接头允许引物在延伸后被切割，以获得在质谱仪上更容易检测到的短DNA片段。
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公开(公告)号：US10006076B2
公开(公告)日：2018-06-26
申请号：US14113354
申请日：2011-07-29
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.
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公开(公告)号：US09275188B2
公开(公告)日：2016-03-01
申请号：US13005863
申请日：2011-01-13
申请人： Markus Kostrzewa
发明人： Markus Kostrzewa
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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