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Method and device for detecting bacteria

A technology to detect bacteria and to be detected, which is applied in the accelerated and highly sensitive detection of bacterial pathogens, in the field of bacteria-phage-complex, which can solve the problems of food recall, economic loss, prolonged storage time, etc., and achieve easy processing, cost reduction, The effect of highly sensitive identification

Inactive Publication Date: 2021-02-02
SINAMIRA AG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] On the other hand, foodborne bacterial food poisoning and food toxin infection is a global problem and causes huge economic losses
The long period until reliable detection of pathogens is obtained implies considerable financial losses due to prolonged storage of the product until resale or the corresponding food may even be recalled

Method used

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  • Method and device for detecting bacteria
  • Method and device for detecting bacteria
  • Method and device for detecting bacteria

Examples

Experimental program
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Effect test

Embodiment 1

[0175] Will 1×10 11 pfu previously purified and concentrated TB54 phage (see above) was resuspended in a solution containing 0.1M NaHCO 3 10ml of PBS (pH 8.5). Subsequently, every 1 x 10 10 Add 100 nmol 5(6)-carboxyfluorescein NHS ester to pfu. Coupling reactions were performed at room temperature in the dark for 1 hour. Such as Figure 7 Schematically shown in , the NHS ester of 5(6)-carboxyfluorescein (2) reacts with primary amino groups of proteins on the surface of TB54 bacteriophage (1) to form stable amide bonds.

[0176] Conjugates were purified by precipitation or by chromatographic methods as described above. Subsequently, the conjugate was photophysically characterized by UV / Vis and fluorescence spectroscopy.

[0177] exist Figure 8 , the excitation and emission spectra of 5(6)-carboxyfluorescein-labeled TB54 phage are illustrated. The spectra show the excitation and emission maxima of 5(6)-carboxyfluorescein at a wavelength of 495 nm and in the range of 520...

Embodiment 2

[0179] A solution of fluorescein isothiocyanate (FTIC) in 0.1 M carbonate buffer (pH 9.0) was prepared. Appropriate aliquots were added to the phage solution prepared in Example 1 and mixed carefully. The phage suspension was incubated in the dark for 1 hour at room temperature. Purification and characterization of the conjugate was performed as described in Example 1.

[0180] 3. Specificity of Bacteria-Phage Interactions

[0181]To demonstrate the great host specificity of the test phage of the tagged species, in a preparatory step, strain C600 (PTC PhageTechnology Center GmbH) was prepared in 10 ml TSB (Tryptic Soy Broth) (Sigma-Aldrich) under standard conditions at 37°C. ), ECOR34 (PTC Phage Technology Center GmbH) and XL10-Gold (Agilent) E. coli bacteria overnight culture. The next day overnight cultures were diluted (1:10 in TSB) and incubated for an additional 2 hours at 37°C. The bacterial suspensions obtained in this way were then mixed in equal proportions (1:1:1...

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Abstract

The present invention relates to a rapid, simple and very sensitive method for the detection of bacteria comprising the steps of: providing one or more suspensions each comprising at least one species of labeled test phage, said The labeled test phage specifically binds the bacterial species to be detected; adding a sample to be tested for the presence of at least one bacterial species to be detected is added to the one or more suspensions; filtering the reaction mixture; testing the retentate Bacteria-phage-complexes on the filter surface, provided that there is at least one bacterial species to be detected, wherein said complex consists of bacteria of said at least one bacterial species to be detected and at least one of said at least one bacterial species bound to it The test phage composition of a kind of test phage; the unbound test phage in the filtrate is detected; the processor assists in processing the received detection signal and outputs the detection result. Furthermore, reaction vessels and measuring instruments for carrying out the method are disclosed.

Description

technical field [0001] The present invention relates to a method for the detection of bacteria, in particular an accelerated and highly sensitive method for the detection of bacterial pathogens, which is based on the formation of specific bacteria-phage-complexes. In addition, reaction vessels and measuring devices for carrying out the method of the invention are disclosed. Background technique [0002] Bacteria are often considered the causative agents of serious disease. In particular, strains of multidrug-resistant bacteria, also known as "nosocomial pathogens", are responsible for a large number of bacterial infections in hospitals. Especially in light of demographic changes and the possibilities of modern intensive care medicine, an increasing number of elderly patients who are particularly susceptible to bacterial infections are found in intensive care units. One of the important components of intensive care therapy is microbiological diagnosis. Rapid and correct id...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N21/00B01L3/00
CPCB01L3/502B01L3/545G01N33/56911B01L2300/021B01L2300/022B01L2300/0654B01L2300/0681B01L2300/0816B01L2300/0864Y02A50/30B01D29/00C12Q1/04C12Q1/70G01N1/28B01L3/502715
Inventor W·米德勒O·弗雷德里希D·吉伯特
Owner SINAMIRA AG
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