DE19538768A1 - Detection of microbial contamination, e.g. in food, drink and water - Google Patents
Detection of microbial contamination, e.g. in food, drink and waterInfo
- Publication number
- DE19538768A1 DE19538768A1 DE1995138768 DE19538768A DE19538768A1 DE 19538768 A1 DE19538768 A1 DE 19538768A1 DE 1995138768 DE1995138768 DE 1995138768 DE 19538768 A DE19538768 A DE 19538768A DE 19538768 A1 DE19538768 A1 DE 19538768A1
- Authority
- DE
- Germany
- Prior art keywords
- sample
- intensity
- photon emission
- detection
- measured
- 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.)
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Links
Classifications
-
- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
Abstract
Description
Die Erfindung betrifft ein Verfahren zum Nachweis mikrobieller Infektion.The invention relates to a method for detection microbial infection.
Bisher hat es sich als nützlich erwiesen, Bakterien oder mikrobielle Infektionen in Flüssigkeiten (zum Beispiel Bier) durch Koloniebildung der möglichen Keime in geeigneten Nährmedien nachzuweisen. Dabei werden die Proben auf Nährböden angesetzt und unter günstigen Wachstumsbedingungen beobachtet. Läßt sich bei hinreichend großer Anzahl von Proben nach einer genügend langen Inkubationszeit von einigen Tagen kein Keimwachstum feststellen, geht man davon aus, daß keine Keime vorhanden sind. Diese Methode wird heute praktisch überall dort eingesetzt, wo der Nachweis mikrobieller Infektionen verlangt wird. Eine ausführliche Beschreibung dieses gebräuchlichen Verfahrens findet man in einschlägigen Lehrbüchern, beispielsweise bei A. Koch, Growth Measurements in: Manual of Methods for General Bacteriology, (ed by Gerhardt, Murray, Costilow, Nester, Wool, Krieg and Phillips) American Society for Microbiology, 1981 pp. 179-206.So far, it has proven useful, bacteria or microbial infections in liquids (e.g. beer) through colony formation of the possible germs in suitable Detect culture media. Doing the samples Culture media set up and under favorable growth conditions observed. With a sufficiently large number of Samples after a sufficiently long incubation period of a few No germ growth observed for days, it is assumed that there are no germs. This method is used today used practically wherever proof microbial infections is required. A detailed one A description of this common method can be found in relevant textbooks, for example by A. Koch, Growth Measurements in: Manual of Methods for General Bacteriology, (ed by Gerhardt, Murray, Costilow, Nester, Wool, Krieg and Phillips) American Society for Microbiology, 1981 pp. 179-206.
Dieses gebräuchliche Verfahren bietet relativ hohe Sicherheit, hat aber den großen Nachteil, daß die Inkubationszeit bis zum sicheren Nachweis vorhandener Bakterien oft länger dauert als es sich ein Hersteller keimfreier Produkte erlauben kann. Die produzierte Ware kann deshalb aus technischen oder wirtschaftlichen Gründen nicht mehr vor der Auslieferung auf Keimfreiheit überprüft werden.This common method offers relatively high ones Security, but has the major disadvantage that the Incubation time until reliable detection of existing Bacteria often take longer than a manufacturer germ-free products. The goods produced can therefore not for technical or economic reasons be checked for sterility before delivery.
In den letzten Jahren haben sich deshalb Fluoreszenz- Verfahren eingebürgert, die den Umstand nutzen, daß Bakterien durch chemische Stoffe oder durch biochemische Eingriffe zur Fluoreszenz angeregt werden können. Dies erlaubt den direkten Nachweis bestimmter Bakterien ohne zeitliche Verzögerung. Die Methode ist vollständig in der Literatur beschrieben, so zum Beispiel in: Wolff, L.F., Anderson, L., Sandberg, G.P., Reither, L., Binsfeld, C.A., Corinaldesi, G., and Shelburne, C.E.: Bacteria Concentration Fluorescence Immunoassay (BCFIA) for the Detection of Periodontopathogens in plaque. J. Periodontol. 1992, 63.1093-1101.For this reason, fluorescence Naturalized procedures that take advantage of the fact that bacteria through chemical substances or through biochemical interventions Fluorescence can be excited. This allows direct Detection of certain bacteria without delay. The Method is fully described in the literature, so for Example in: Wolff, L.F., Anderson, L., Sandberg, G.P., Reither, L., Binsfeld, C.A., Corinaldesi, G., and Shelburne, C.E .: Bacteria Concentration Fluorescence Immunoassay (BCFIA) for the detection of periodontopathogens in plaque. J. Periodontol. 1992, 63.1093-1101.
Die Fluoreszenzmethode kann zwar ohne Zeitverzögerung Bakterien unmittelbar nachweisen, hat aber den Nachteil, daß sie nur für spezielle Bakterien, die sich biochemisch zur Fluoreszenz anregen lassen, tauglich ist. Darüberhinaus liegt die Nachweisgrenze selbst in günstigen Fällen bei 10⁴ Bakterien/ml. Sie ist deshalb nicht generell einsetzbar, sondern relativ individuell und kostspielig.The fluorescence method can be used without a time delay Detect bacteria immediately, but has the disadvantage that they only for special bacteria that are biochemically responsible for Let fluorescence be stimulated. Beyond that the detection limit even in favorable cases at 10⁴ Bacteria / ml. It is therefore not generally applicable but relatively individual and expensive.
Aus den Patenten (Europäisches Patent EP 0430150, Flüssigkeiten DE 44 01 169 A1, Zellkulturen P 43 08 520.2-41) ist ein Verfahren bekannt, das die Photonenemission bis zu einer Empfindlichkeit von 10-17W (entsprechend einiger weniger Quanten pro Sekunde und pro cm²) im optischen Spektralbereich (von ca. 200-800 nm) erlaubt. Mithilfe dieses Verfahrens kann auch das Rekombinationsleuchten von Ladungsträgern in Flüssigkeiten gemessen werden.From the patents (European patent EP 0430150, liquids DE 44 01 169 A1, cell cultures P 43 08 520.2-41) a method is known that the photon emission up to a sensitivity of 10 -17 W (corresponding to a few quanta per second and per cm²) in the optical spectral range (from approx. 200-800 nm) allowed. This method can also be used to measure the recombination lighting of charge carriers in liquids.
Überraschend zeigte sich nun, daß dieses Verfahren auch geeignet ist, mindestens 100 Bakterien/ml in polaren Flüssigkeiten anzuzeigen. Obwohl bereits die hohe Sensitivität des Verfahrens zur Unterscheidung der "Qualität" von Flüssigkeiten als überraschend anzusehen war, lieferten neue Untersuchungen zum Nachweis mikrobieller Infektion den zusätzlich überraschenden Befund, daß alle mikrobiellen Infektionen auch schon bei so geringen Konzentrationen, daß sie mit den anderen Methoden nicht mehr nachweisbar sind, durch signifikante Veränderungen der Photonenabstrahlung auffallen. Dabei hat es sich als vorteilhaft erwiesen, die Probe in einer wäßrigen Lösung in eine 10 ml Quarzglasküvette zu bringen. Um Ladungsträger in die Flüssigkeit zu bringen, kann man vorteilhaft mit 3 mM/l Kochsalz anreichern. An zwei Platinelektroden, die nadelförmig parallel im Abstand von einigen Millimetern in die Flüssigkeit getaucht werden, legt man beispielsweise eine Gleichspannung von 30 Volt, die nach einigen Sekunden wieder abgeschaltet wird. Die Probe befindet sich in absoluter Dunkelheit. Die Intensität der Photonenemission wird mit einem Lichtmeßgerät (das als Gebrauchsmuster beispielsweise in G 94 17 845.3 beschrieben ist) gemessen.Surprisingly, it has now been found that this method too is suitable, at least 100 bacteria / ml in polar Display liquids. Although already the high Sensitivity of the process to differentiate the "quality" of liquids was surprising new investigations to detect microbial infection additionally surprising finding that all microbial Infections even at such low concentrations that they are no longer detectable with the other methods, through significant changes in photon radiation stand out. It has proven to be advantageous that Sample in an aqueous solution in a 10 ml quartz glass cuvette bring to. In order to bring charge carriers into the liquid, can advantageously be enriched with 3 mM / l sodium chloride. On two Platinum electrodes that are needle-shaped parallel at a distance of a few millimeters into the liquid for example, a DC voltage of 30 volts, according to is switched off again for a few seconds. The sample is located yourself in absolute darkness. The intensity of the Photon emission is with a light meter (which as Utility model is described for example in G 94 17 845.3) measured.
Es zeigt sich, daß die Anwesenheit mikrobieller Infektion die Photonenemission empfindlich verändert.It turns out that the presence of microbial infection Sensitive change in photon emission.
Durch geeignete Variation der Meßparameter (Spannung, pH- Wert, Zusammensetzung der Flüssigkeit, externe Anregung, spektrale Auflösung) läßt sich die Art und der Zustand der Infektion bestimmen, beispielsweise die Frage, um welche Bakterien es sich handelt und in welchem Zustand (lebendig oder tot) die Bakterien vorliegen. Deshalb werden neben den ersten beiden Patentansprüchen auch die Ansprüche 3.-7. erhoben.By suitable variation of the measurement parameters (voltage, pH Value, composition of the liquid, external excitation, spectral resolution), the type and state of the Determine infection, for example the question of which Bacteria and in what condition (alive or dead) the bacteria are present. Therefore, in addition to the the first two claims also claims 3-7. raised.
Die Erfindung ist überraschend und neuartig, da die hohe Empfindlichkeit des Verfahrens trotz der bekannten und teilweise patentierten Grundbausteine nicht bekannt oder vorhersehbar war. Diese hohe Empfindlichkeit bei gleichzeitig hinreichender Reproduzierbarkeit macht das Verfahren für viele gewerbliche Gebiete interessant.The invention is surprising and novel because of the high Sensitivity of the method despite the known and partially patented basic building blocks not known or was predictable. This high sensitivity at the same time sufficient reproducibility makes the process for many commercial areas interesting.
Das Verfahren ist einzusetzen in der Getränke- und Brauwirtschaft, bei der Überwachung von Wasser, in der Lebensmittelindustrie, wie beispielsweise bei Milchprodukten und auch in allen anderen Zweigen der gewerblichen Wirtschaft, die auf die Kontrolle mikrobieller Infektionen angewiesen sind.The method is used in the beverage and Brewing industry, in the monitoring of water, in the Food industry, such as dairy products and also in all other branches of the commercial Economy based on the control of microbial infections are instructed.
In eine 10 ml Quarzküvette werden zwei nadelförmige Platinelektroden in einem Abstand von 5 mm parallel eingebracht und mit einer Gleichspannungsquelle verbunden. In die Küvette bringt man hintereinander je 8 ml reine Kochsalzlösung (3 mM/l Kochsalz), die gleiche Lösung mit zusätzlich einer Konzentration von 10 Rhizobium japonicum 1132-2 Bakterien/ml und die gleiche Lösung mit zusätzlich einer Konzentration von 100 Rhizobium japonicum 1132-2 Bakterien/ml.Two needle-shaped are placed in a 10 ml quartz cuvette Platinum electrodes at a distance of 5 mm in parallel introduced and connected to a DC voltage source. In the cuvette is brought 8 ml pure Saline (3 mM / l saline), the same solution with additionally a concentration of 10 Rhizobium japonicum 1132-2 bacteria / ml and the same solution with additional a concentration of 100 Rhizobium japonicum 1132-2 Bacteria / ml.
In jedem Fall wird für eine Zeit von 5 Sekunden eine Gleichspannung von 80 Volt angelegt. Gleichzeitig werden die Intensitäten der Photonenemission (in Anzahl der Photonen/100 ms) über den Zeitraum von 5 s gemessen. Die Messung wird dreimal wiederholt. Es werden die Mittelwerte und Streuungen der drei Messungen gebildet. Tabelle I enthält die Ergebnisse.In any case, for a time of 5 seconds DC voltage of 80 volts applied. At the same time, the Intensities of the photon emission (in number of Photons / 100 ms) measured over a period of 5 s. The Measurement is repeated three times. It will be the averages and scatter of the three measurements. Table I contains the results.
Das Ergebnis zeigt, daß man mit dieser Methode signifikant noch 10 Bakterien/ml nachweisen kann.The result shows that you can use this method significantly can still detect 10 bacteria / ml.
Claims (7)
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19538768A DE19538768C2 (en) | 1995-10-18 | 1995-10-18 | Procedure for the detection of a microbial infection |
CN 96105260 CN1072799C (en) | 1995-10-18 | 1996-05-28 | Sensitive method for quickly testing microbe pollution by electroluminescence and its use |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19538768A DE19538768C2 (en) | 1995-10-18 | 1995-10-18 | Procedure for the detection of a microbial infection |
Publications (2)
Publication Number | Publication Date |
---|---|
DE19538768A1 true DE19538768A1 (en) | 1997-04-24 |
DE19538768C2 DE19538768C2 (en) | 2003-02-27 |
Family
ID=7775154
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE19538768A Expired - Fee Related DE19538768C2 (en) | 1995-10-18 | 1995-10-18 | Procedure for the detection of a microbial infection |
Country Status (2)
Country | Link |
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CN (1) | CN1072799C (en) |
DE (1) | DE19538768C2 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN106085833B (en) * | 2016-05-31 | 2018-03-13 | 华北理工大学 | Microorganism metering device and its metering method in water |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4283490A (en) * | 1978-07-28 | 1981-08-11 | Plakas Chris J | Method for detection of low level bacterial concentration by luminescence |
DE3038255A1 (en) * | 1980-10-10 | 1982-05-19 | Wolfgang Prof. 7500 Karlsruhe Mehlhardt | Examining biological effects on foodstuffs of seeds - by measuring intensity of ultra-weak photon radiation in vitro |
DE3040855A1 (en) * | 1980-10-30 | 1982-06-09 | Wolfgang Prof. 7500 Karlsruhe Mehlhardt | Examining biological effects on foodstuffs of seeds - by measuring intensity of ultra-weak photon radiation in vitro |
US4421848A (en) * | 1980-04-15 | 1983-12-20 | Whitlock Gerald D | Method of detecting the presence of live organisms in substances |
DE3737649A1 (en) * | 1987-11-06 | 1989-05-24 | Inst Zellforschung Und Biolumi | Method for determining the luminescence of cell cultures, and device for carrying out the method |
DE3935974A1 (en) * | 1989-10-28 | 1991-05-02 | Mueller Klieser Wolfgang Prof | Determn. of spatial distribution of metabolites in tissue samples - by bio-luminescence, using conc. viscous soln. of luciferase and counting photons in unit area |
EP0430150A2 (en) * | 1989-11-29 | 1991-06-05 | Fritz-Albert Popp | Method for testing quality and quality changes of biological systems and organochemical compositions interacting with these systems using measurements of ultraweak photon emission |
DE4308520A1 (en) * | 1993-03-17 | 1994-09-22 | Popp Fritz Albert Dr | Method for differentiating between homozygotes, heterozygotes and normal cells of an organism |
DE4401169A1 (en) * | 1994-01-17 | 1995-07-20 | Popp Fritz Albert Dr | Faster procedure for detecting differences in fluid characteristics |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE9417845U1 (en) * | 1994-11-08 | 1995-04-20 | Popp Fritz Albert Dr | System for measuring ultra-weak photon emission (photon measuring system) |
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1995
- 1995-10-18 DE DE19538768A patent/DE19538768C2/en not_active Expired - Fee Related
-
1996
- 1996-05-28 CN CN 96105260 patent/CN1072799C/en not_active Expired - Fee Related
Patent Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4283490A (en) * | 1978-07-28 | 1981-08-11 | Plakas Chris J | Method for detection of low level bacterial concentration by luminescence |
US4421848A (en) * | 1980-04-15 | 1983-12-20 | Whitlock Gerald D | Method of detecting the presence of live organisms in substances |
DE3038255A1 (en) * | 1980-10-10 | 1982-05-19 | Wolfgang Prof. 7500 Karlsruhe Mehlhardt | Examining biological effects on foodstuffs of seeds - by measuring intensity of ultra-weak photon radiation in vitro |
DE3040855A1 (en) * | 1980-10-30 | 1982-06-09 | Wolfgang Prof. 7500 Karlsruhe Mehlhardt | Examining biological effects on foodstuffs of seeds - by measuring intensity of ultra-weak photon radiation in vitro |
DE3737649A1 (en) * | 1987-11-06 | 1989-05-24 | Inst Zellforschung Und Biolumi | Method for determining the luminescence of cell cultures, and device for carrying out the method |
DE3935974A1 (en) * | 1989-10-28 | 1991-05-02 | Mueller Klieser Wolfgang Prof | Determn. of spatial distribution of metabolites in tissue samples - by bio-luminescence, using conc. viscous soln. of luciferase and counting photons in unit area |
EP0430150A2 (en) * | 1989-11-29 | 1991-06-05 | Fritz-Albert Popp | Method for testing quality and quality changes of biological systems and organochemical compositions interacting with these systems using measurements of ultraweak photon emission |
DE3939411A1 (en) * | 1989-11-29 | 1991-06-06 | Popp Fritz Albert | METHOD FOR THE EXAMINATION OF THE QUALITY AND QUALITY CHANGES OF BIOLOGICAL SYSTEMS AND INTERACTIVE ORGANIC CHEMICAL COMPOUNDS THEREOF BY MEASUREMENT OF THE ULTRA-CHANGING PHOTON EMISSION |
DE4308520A1 (en) * | 1993-03-17 | 1994-09-22 | Popp Fritz Albert Dr | Method for differentiating between homozygotes, heterozygotes and normal cells of an organism |
DE4401169A1 (en) * | 1994-01-17 | 1995-07-20 | Popp Fritz Albert Dr | Faster procedure for detecting differences in fluid characteristics |
Also Published As
Publication number | Publication date |
---|---|
CN1072799C (en) | 2001-10-10 |
DE19538768C2 (en) | 2003-02-27 |
CN1150246A (en) | 1997-05-21 |
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Owner name: BUEHLER AG, UZWIL, CH |
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Effective date: 20110502 |