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EP1147386A1 - Detecteur pour micro-organismes - Google Patents

Detecteur pour micro-organismes

Info

Publication number
EP1147386A1
EP1147386A1 EP00914026A EP00914026A EP1147386A1 EP 1147386 A1 EP1147386 A1 EP 1147386A1 EP 00914026 A EP00914026 A EP 00914026A EP 00914026 A EP00914026 A EP 00914026A EP 1147386 A1 EP1147386 A1 EP 1147386A1
Authority
EP
European Patent Office
Prior art keywords
organisms
micro
optical
fluctuations
microorganisms
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00914026A
Other languages
German (de)
English (en)
Inventor
Vladimir Agueev
Olga Agueev
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mediquant GmbH
Original Assignee
Mediquant GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from RU99101910/13A external-priority patent/RU99101910A/ru
Application filed by Mediquant GmbH filed Critical Mediquant GmbH
Publication of EP1147386A1 publication Critical patent/EP1147386A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/21—Polarisation-affecting properties
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics

Definitions

  • the invention relates to measurement technology and in particular to optical biosensors. Microorganisms in liquids are currently recorded using optical microscopy, bioluminescence and selective coloring. The common disadvantage of all of these methods and their metrological implementation is the low degree of automation of the measuring process.
  • a device (prototype) for recording mobile microorganisms (BM) which consists of a laser radiation source, a cuvette and a photo receiver (FE), which are optically coupled to one another [1].
  • the spectrum of the intensity fluctuations of the scattered radiation is recorded. This spectrum is different for Brownian particles (BT) and for BM.
  • the disadvantage of this method is its low selectivity in the detection of the microorganisms against the background of the scatter signal of the Brownian particles.
  • UM immobile microorganisms against the background of Brownian particles in liquid is not possible with this method, since the fluctuation scatter spectra of BT and UM do not differ.
  • Another shortcoming of this method is its low sensitivity, which is related to the fact that the useful signal is superimposed by the noise of the laser beam itself, which is particularly noticeable when using non-stabilized lasers.
  • the photo receivers are each optically coupled to their associated cuvette and are illuminated by a common laser radiation source observed, the spectra of the scattering intensity fluctuation in the two cuvettes, and the difference is an indication of whether '.n one of the two cuvettes BM present or only if they have different activity in the cells.
  • the aim of the invention is to develop an optical variant of the device which makes it possible to record mobile as well as immobile living microorganisms in liquid against the background of a large number of Brownian particles. This is achieved by recording both the movement (for BM) and the course of the metabolic processes (for UM and BM).
  • the reliability of the detection of individual microorganisms and the improvement in sensitivity are achieved by increasing the examination volume and suppressing the noise of the radiation source.
  • the essence of the invention is that a linearly polarized examination radiation penetrates the liquid to be examined in the cuvette, the examination volume being the same as the cuvette volume, and the fluctuations in the optical activity of the medium with BM and UM being recorded using two possible methods. In the first method, a combined polarizer and analyzer (at an extinction angle) and a photo receiver are arranged in the light beam that penetrates the examination volume.
  • the signal from the photo receiver is sent to an amplifier that has a large gain factor, but only amplifies the variable part of the signal and does not record the constant part. This distinguishes it from the generally known devices used in saccharometry [3]. If there are no optically active particles in the examination volume or if their concentration does not change over time, then the output signal of the amplifier is zero. If, on the other hand, living microorganisms are present in the examination volume, they lead to stochastic changes in the optical activity of the medium through their life processes and the metabolic processes and organic chemical reactions taking place in them. The amplitude of the fluctuations recorded is proportional to the concentration of the microorganisms and the intensity of the metabolic processes taking place in them, which is e.g.
  • the device requires a light source that is highly stabilized in terms of intensity, since its noise is recorded directly by the photoreceiver and amplified by the amplifier of the variable signal, which considerably reduces the signal-to-noise ratio and accordingly the sensitivity of the device.
  • a differential circuit is used for the recording, in which the radiation, after penetrating the examination volume, is passed to a separating device for orthogonal polarizations, e.g.
  • a glan prism arrives, and the intensity of the two light beams separated by the prism with orthogonal polarizations is recorded by two photo receivers, each of which is optically coupled with their polarization.
  • the two signals to be recorded are applied to different inputs of the differential amplifier (DV), the output of which is coupled to an evaluation unit that determines the mean signal amplitude for a fixed period of time, the dispersion and the Fourier spectrum.
  • the noise of the light source is present at the same time and in phase at both photo receivers and is subtracted from the DV with high accuracy (up to 100 dB).
  • the signal of the intensity fluctuations which is related to the fluctuation of the optical activity, is not only not subtracted from the DV, but even amplified.
  • Figure 1 shows the block diagram of the device.
  • 1 - source of the stabilized linearly polarized optical radiation e.g. a stabilized laser
  • 2 - cuvette with the medium to be examined
  • 3 - polarizer and analyzer
  • 4 - photo receiver
  • 5 - variable signal amplifier
  • 6 - evaluation and display unit
  • Figure 2 shows the same block diagram, but with a differential circuit for recording.
  • Figure 3 shows the dependence of the mean fluctuation amplitude on the time for the rehydrant of the immobile microorganisms Saccharomyces cerevisiae, which were introduced into the test volume with initially sterile water with glucose. It can be seen that there are no fluctuations and consequently no metabolic processes in the course of about 20 minutes, which would be necessary for the activation of the spores of the microorganisms and an increase in the signal after the start of the active phase.
  • Figure 4 shows the dependence of the dispersion of the fluctuation amplitude on the time before and after heating the test volume to 80 ° C. There is an abrupt decrease in the dispersion after heating, which is related to the partial death of the microorganisms and the reduction in the metabolic processes.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Immunology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Toxicology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

L'invention concerne une technique de mesure et notamment des biocapteurs optiques. Elle peut être employée par exemple pour déterminer l'efficacité de différents antibiotiques sur divers micro-organismes pathogènes ou pour contrôler la qualité de l'eau. Le dispositif permet de détecter des micro-organismes mobiles ou immobiles dans l'eau par des techniques optiques. Celles-ci permettent de déterminer les fluctuations de l'activité optique du milieu, qui sont liées aussi bien à la mobilité des micro-organismes qu'à leurs processus métaboliques. Afin d'améliorer la sensibilité de mesure, on utilise un circuit différentiel qui permet de supprimer le bruit de fond du rayonnement servant à l'analyse.
EP00914026A 1999-02-05 2000-02-02 Detecteur pour micro-organismes Withdrawn EP1147386A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
RU99101910 1999-02-05
RU99101910/13A RU99101910A (ru) 1999-02-05 Детектор микроорганизмов
PCT/DE2000/000343 WO2000046579A1 (fr) 1999-02-05 2000-02-02 Detecteur pour micro-organismes

Publications (1)

Publication Number Publication Date
EP1147386A1 true EP1147386A1 (fr) 2001-10-24

Family

ID=20215316

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00914026A Withdrawn EP1147386A1 (fr) 1999-02-05 2000-02-02 Detecteur pour micro-organismes

Country Status (2)

Country Link
EP (1) EP1147386A1 (fr)
WO (1) WO2000046579A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IL145683A0 (en) * 2001-09-26 2002-06-30 Enoron Technologies Ltd Apparatus and method for measuring optically active materials

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8625530D0 (en) * 1986-10-24 1986-11-26 Goodall D M Optical apparatus
US4902134A (en) * 1988-02-03 1990-02-20 Rudolph Research Corporation Optical amplifier and method for amplifying optical polarization state change effects
US5268305A (en) * 1989-06-15 1993-12-07 Biocircuits Corporation Multi-optical detection system
US5398681A (en) * 1992-12-10 1995-03-21 Sunshine Medical Instruments, Inc. Pocket-type instrument for non-invasive measurement of blood glucose concentration

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0046579A1 *

Also Published As

Publication number Publication date
WO2000046579A1 (fr) 2000-08-10

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