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Analysis of microbes by MALDI mass spectrometry

A technology of mass spectrometry and microbiology, which is applied in the field of mass spectrometers with matrix-assisted laser desorption ionization, can solve problems such as the influence of identification reliability, and achieve the effects of saving culture time, avoiding errors, and reducing costs

Active Publication Date: 2015-02-25
BRUKER DALTONIK GMBH & CO KG
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Problems solved by technology

Identification reliability is only slightly affected

Method used

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  • Analysis of microbes by MALDI mass spectrometry
  • Analysis of microbes by MALDI mass spectrometry
  • Analysis of microbes by MALDI mass spectrometry

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Embodiment Construction

[0040]There are 4 stages of microbial growth. During an initial short adjustment period ("lag phase"), the microorganisms adapt their metabolism to the medium by eg producing different digestive enzymes. This is followed by an unimpeded growth phase ("log phase") in which the microorganisms grow exponentially until nutrient shortages or production of toxins stop the exponential growth angle and enter a "plateau phase" asymptomatically. Finally, the microorganisms die ("death phase") due to lack of nutrients or poisoning. During unhindered growth, bacterial cells divide after each characteristic period ("generation time"). The generation time depends on the microbial species as well as the external conditions; for optimal conditions (temperature, medium), the generation time in each case ranged between 15 minutes and 24 hours. Most clinically important pathogens divide within a focal period of 15 to 45 minutes; however, there are also clinically important pathogens that grow ...

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Abstract

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 (MALDl). The objective of the invention is to significantly shorten the mass spectrometric identification method for diagnostically important microbes. To achieve this, 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 MALDl samples. The ionization by matrix-assisted laser desorption also takes place on the contact surface. With a preparation that produces a high ion yield in the MALDl process, the critical period from delivery of the samples for analysis until the identification of the microbes can be reduced from around 18 to 24 hours to around only eight hours for most of the clinically relevant microbes.

Description

technical field [0001] The invention relates to a method for using mass spectrometry to analyze microorganisms in bacterial colonies on the surface of a culture medium, in particular to a mass spectrometer using matrix-assisted laser desorption ionization (MALDI: ionization by matrix-assisted laser desorption). Background technique [0002] Routine fast and error-free microbiological analysis plays an important role in clinical and extraclinical infection diagnosis, sanitation monitoring in hospitals or swimming rivers or lakes, food analysis, biotechnological process monitoring and control, and microbiological research. "Microbial species" here may be simply referred to as microorganisms, usually small microscopic organisms such as single-celled fungi (such as yeast), microscopic algae, protozoa (such as the malaria-causing Plasmodium), although identification efforts usually focus on bacteria. [0003] Identification of microorganisms means, in principle, the determinatio...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569H01J49/00
CPCG01N33/569H01J49/0027H01J49/164
Inventor 彼得·拉什迪特尔·瑙曼马伦·斯坦勒托马斯·迈尔延斯·波斯迈尔
Owner BRUKER DALTONIK GMBH & CO KG
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