Bruker Daltonik Gmbh

Other

Daimler Chrysler Ag

Thomas Maier

Markus Kostrzewa

Jochen Franzen

Karsten Michelmann

Norman Mauder

Wolfgang Pusch

Gert Volk

Wolfram Thomas

Stefan Klepel

Jurgen Trost

Peter Lasch

Dieter Naumann

Martin Rothmund

Jens Boβmeyer

Thomas Kind

Jens Bossmeyer

Erwin Schmid

J&uuml;rgen Trost

Richard Gall

Thilo Schauer

Maren Stämmler

Michael Maier

Detlef Beckmann

G&uuml;nter Ahlert

The invention relates to a microbial test standard for use in infrared spectrometry which has at least two resealable vessels which are liquid-tight when closed, each of which contains a predefined amount of dried biomass of a microorganism. The microorganisms in the different vessels differ in at least one characteristic, which is selected in particular from the group comprising species, subspecies, strain, serovar, pathovar, toxivar and variety, and the difference manifests itself in a predefined intermicrobial spectral distance. The disclosure furthermore comprises a method of using the microbial test standard.

Microbial test standard for use in infrared spectrometry

The invention mainly relates to the mass spectrometric identification of pathogens in blood cultures from bloodstream 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.

Mass spectrometric diagnosis of septicemia

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.

Mass spectrometric diagnosis of sepsis without blood culture

The invention relates to methods for the mass spectrometric analysis of microbes, in particular to a transfer method of microbes that are required to be identified from agar plates onto mass spectrometric sample supports and their preparation for ionization by matrix-assisted laser desorption (MALDI). Microbes from microcolonies which have grown on the agar plates after a culture time of only six to eight hours are transferred with a high transfer yield onto contact surfaces of suitable size by direct contact and cell disrupted on the contact surface; the released proteins are prepared with matrix material on the contact surface as MALDI samples. The ionization by matrix-assisted laser desorption also takes place on the contact surface.

Analysis of microbes by MALDI mass spectrometry

A method is provided for identifying a hyphal fungus in a sample by similarity comparisons between a mass spectrum of the fungus and reference spectra. The method includes growing fresh hyphae without any adhering contact to surfaces as to form a mycelium with undifferentiated hyphae cells. A sample is prepared from the hyphae cells, and a mass spectrum is acquired of the hyphae cell sample. The mass spectrum is matched with at least one reference spectrum.

Mass spectrometric identification of hyphal fungi

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.

Acquisition technique for MALDI time-of-flight mass spectra

A simplified sample preparation on a sample support for ionization with laser desorption (MALDI or LDCI, for example) includes depositing an analyte material onto a deposition site, providing a sample site which is intended as the substrate for a matrix crystal layer at a distance from the deposition site, and establishing a liquid communication between the deposition site and the sample site. A device for the preparation of samples for ionization with matrix-assisted laser desorption, and a sample support for use with the inventive method are also disclosed.

Sample preparation for ionization with matrix-assisted laser desorption

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.

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.

Spectrophotometric identification of microbe subspecies

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.

Mass scale alignment of time-of-flight mass spectra

A method of adapting image information to the perceptive capacity of the human eye, having the following steps: (a) displaying lightness values and colorimetric values or chromaticity steps from an original image as a group of points in an initial color space; (b) transferring the group of points into a physiological substantially equal-spaced perceived color space while maintaining the geometry of the group of points within the limits of an output color space projected into the perceived color space; (c) transforming the group of clouds from the perceived color space into the output color space by using the transformation equations existing between the perceived color space and the output color space; and/or (d) displaying the image with lightness values and colorimetric values or chromaticity steps in accordance with the points contained in the output color space.

Method of registering image information

A single processor system features independent multiple watchdog units allocated to a processor unit of the system, the watchdog units operable to monitor for system faults and, upon detecting a fault, further operative to place the processor system into a predetermined fault reaction state. The multiple watchdog units are each provided with respective different watchdog calls which are output to the processor unit depending upon program execution.

Single-processor system

An eating utensil with a spoon shell fastened to a handle enables handicapped people, who have, e.g., only one usable hand, to reduce in size pieces of food, e.g., meat, by cutting into bite-sized pieces. To achieve this, the spoon shell ( ) and the handle ( ) are of a two-part design. The spoon shell ( ) has a first shell part ( ) with a first handle shaft ( ) and a second shell part ( ) with a second handle shaft ( ). The respective shell parts ( ) and handle shafts ( ), which are rigidly connected to one another, are connected to one another by a scissor hinge ( ) and can be moved against one another in a scissor-like manner. The shell parts ( ) have cutting edges ( ) cooperating in a scissor-like manner.

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Lilienthal, Germany

Thomas Maier