DD146769A3 - METHOD FOR TESTING THE CHEMOTHERAPEUTIC SENSITIVITY OF BACTERIA - Google Patents
METHOD FOR TESTING THE CHEMOTHERAPEUTIC SENSITIVITY OF BACTERIA Download PDFInfo
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- DD146769A3 DD146769A3 DD76195937A DD19593776A DD146769A3 DD 146769 A3 DD146769 A3 DD 146769A3 DD 76195937 A DD76195937 A DD 76195937A DD 19593776 A DD19593776 A DD 19593776A DD 146769 A3 DD146769 A3 DD 146769A3
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- German Democratic Republic
- Prior art keywords
- bacteria
- chemotherapeutic
- testing
- sensitivity
- chemotherapeutic sensitivity
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- 238000012360 testing method Methods 0.000 title claims abstract description 13
- 238000000034 method Methods 0.000 title claims abstract description 11
- 230000035945 sensitivity Effects 0.000 title claims abstract description 10
- 230000000973 chemotherapeutic effect Effects 0.000 title claims abstract description 9
- 241000894006 Bacteria Species 0.000 title claims abstract description 8
- 230000001580 bacterial effect Effects 0.000 claims description 8
- 239000000725 suspension Substances 0.000 claims 1
- 239000002246 antineoplastic agent Substances 0.000 abstract description 15
- 229940127089 cytotoxic agent Drugs 0.000 abstract description 14
- 101100072620 Streptomyces griseus ind2 gene Proteins 0.000 abstract 2
- 229940079593 drug Drugs 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 230000004060 metabolic process Effects 0.000 abstract 1
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000002503 metabolic effect Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 210000002700 urine Anatomy 0.000 description 4
- 238000011534 incubation Methods 0.000 description 3
- 230000000721 bacterilogical effect Effects 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 208000019206 urinary tract infection Diseases 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 201000004538 Bacteriuria Diseases 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 230000000254 damaging effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000013208 measuring procedure Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 230000036651 mood Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/497—Physical analysis of biological material of gaseous biological material, e.g. breath
-
- 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
- C12Q1/18—Testing for antimicrobial activity of a material
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Hematology (AREA)
- Pathology (AREA)
- Toxicology (AREA)
- General Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Beschreibung eines Verfahrens zur Pruefung der chemotherapeutischen Empfindlichkeit von Bakterien, indem das Ergebnis der elektro-chemischen Bestimmung des pO&ind2! oder pCO&ind2! in bakterienhaltigen Loesungen unter Gegenwart eines Chemotherapeutikums als Ausdruck des Umfangs der Stoffwechselbeeinflussung verbindliche Aussagen zur Wahl des geeigneten Medikaments vermittelt.Description of a Method for Testing the Chemotherapeutic Sensitivity of Bacteria by Determining the Result of the Electrochemical Determination of the pO & ind2! or pCO & ind2! in bacteria-containing solutions in the presence of a chemotherapeutic agent as an expression of the extent of the metabolism influencing mediates binding statements on the choice of the appropriate drug.
Description
-4- f 7 J 7 O /-4- f 7 J 7 O /
Titel der Sirf indung:Title of sirf indung:
Elektro-chemisches Verfahren zur Seime Übe Stimmung der chemotherapeutischen Empfindlichkeit pathogener MikroorganismenElectrochemical Process for the Study of Mood of the chemotherapeutic sensitivity of pathogenic microorganisms
Anwendungsgebiet der Erfindung:Field of application of the invention:
Für eine rationelle und vor allem erfolgversprechende Behandlun von bakteriell bedingten Infektionen ist aufgrund des Resistenz Phänomens eine in-vitro-Empfindlichkeitsprüfung des die Erkrankung im Einzelfall verursachenden Bakterienstammes unabdingbar. Dies gilt besonders für Infektionen mit lebensbedrohendem Krankheitsbild bzw, primär chron. rezidivierendenxVerlauf „For a rational and, above all, promising treatment of bacterially induced infections, an in vitro susceptibility testing of the bacterial strain causing the disease in an individual case is indispensable on account of the resistance phenomenon. This is especially true for infections with life-threatening disease or, primarily chron. recurrencexchange "
Als Ergebnis derartiger Voruntersuchungen resultiert ein sog. Antibiogramm, aus dem sich ergibt, welches Chemotherapeutikum die besten Voraussetzungen zur Behandlung der Infektion bietet·As a result of such preliminary investigations, a so-called antibiogram results, which shows which chemotherapeutic agent offers the best conditions for the treatment of the infection.
Die Durchführung dieser Untersuchungen ist ausnahmslos an mikrobiologische Speziallaboratorien gebunden·The execution of these examinations is without exception tied to special microbiological laboratories.
Charakteristik der bekannten technischen Lösungen:Characteristic of the known technical solutions:
Zur Erstellung des Antibiogramms selbst stehen ζ.Zt. 2 verschiedene methodische Prinzipien zur Verfügung.To prepare the antibiogram itself, ζ.Zt. 2 different methodological principles available.
1. Röhrchen- oder Plattenverdünnungsteste: In Reagenzröhrchen mit flüssigem oder in Petrischalen mit feste] Nährboden werden Verdünnungsreihen mit den zu prüfenden Chemotherapeutika angesetzt. Jedes Röhrchen bzw. jede Platte enthält unter Abstufung der Chemotherapeutika-Konzentration die gleiche Menge des vom Patienten isolierten Bakterienstammes. Nach ca. 24 otd. Inkubation bei 37 0C wird die Cheraotherapeutika-Konzentration ermittelt, die zu einer Hemmung des Bakterienwachstums geführt hat. Dabei gilt für jedes Chemotherapeutikum ein Grenzwert, d.h. wird ein Stamm erst durch eine Konzentration gehemmt, die über dieses Limit hinausgeht, muß er im Antibiogramm als resistent gegen das jeweils geprüfte Chemotherapeutikum bezeichnet werden.1. Tube or plate dilution tests: Dilution series with the chemotherapeutic agents to be tested are prepared in test tubes with liquid or in Petri dishes with solid culture medium. Each tube or plate contains the same amount of the bacterial strain isolated from the patient, grading the chemotherapeutic agent concentration. After about 24 otd. Incubation at 37 0 C, the Cheraotherapeutika concentration is determined, which has led to an inhibition of bacterial growth. For each chemotherapeutic agent, a limit value applies, ie if a strain is only inhibited by a concentration which exceeds this limit, it must be described in the antibiogram as resistant to the particular chemotherapeutic agent tested.
-a- 195937-a- 195937
2. Diffusionsteste, fast ausschließlich in Form des 'igardiffusionstests2. Diffusion tests, almost exclusively in the form of the igardiffusion test
Nach Ausspateln des zu prüfenden Bakterienstammes auf einen festen Nährboden mit Agar-Zusatz werden die einzelnen Chemotherapeutika in empirisdh ermittelten Dosen aufgetragen. Als Methode der Wahl'gilt der sog. Blättchentest (mit Chemotherapeutika getränkten Eilterpapierblättchen).After balancing the bacterial strain to be tested on a solid nutrient medium with addition of agar, the individual chemotherapeutic agents are applied in empirically determined doses. As a method of Wahl'gilt the so-called Blättchentest (soaked with chemotherapeutic Eilterpapierblättchen).
Besteht Empfindlichkeit des isolierten Bakterienstammes gegenüber einem bestimmten Chemotherapeutikum, ist nach ca, 24 Std. Inkubation bei 37 0C um das aufgelegte Blättchen eine Wachstumshemmzone mit einem bestimmten Mindestdurchmesser entstanden.If there is sensitivity of the isolated bacterial strain to a particular chemotherapeutic agent, after about 24 hours of incubation at 37 ° C. a growth inhibiting zone having a certain minimum diameter has formed around the applied leaflets.
Beide Verfahren haben neben dem nicht unerheblichen technologischen Zeitaufwand den Nachteil, daß das Ergebnis der Testungen erst frühestens nach 16 - 20 Std. vorliegt, was unter Umständen den Verlauf bakterieller Infektionen in fataler Weise beeinflussen kann·Both methods have the disadvantage, in addition to the not inconsiderable technological time expenditure, that the result of the tests is not present until after 16 to 20 hours at the earliest, which may possibly have a fatal effect on the course of bacterial infections.
Ziel der Erfindung:Object of the invention:
Aufgabe und Inhalt der vorliegenden Erfindung ist die Entwicklung eines Verfahrens, das unter Anwendung eIektro-chemischer Meßprinzipien in wesentlich kürzerer Zeit und ohne den bisher unumgänglichen konventionell-bakteriologischen Aufwand zuverlässige Informationen über die chemotherapeutische Empfindlichkeit pathogener Mikroorganismen liefert·The object and content of the present invention is the development of a method which provides reliable information about the chemotherapeutic sensitivity of pathogenic microorganisms using electrochemical measurement principles in a significantly shorter time and without the previously unavoidable conventional bacteriological effort.
Darlegung des Wesens der Erfindung;Explanation of the essence of the invention;
Grundvoraussetzung der erfindungsgemäßen Lösung dieser Aufgabe ist die in geeigneten Nährmedien jederzeit nachweisbare allgemeine Stoffwechselaktivität von Bakterien. Sie ist u.a. am einfachsten durch Messung des O^-Verbrauchs oder der COp-BiI-dung pro Zeiteinheit feststellbar.The basic requirement of the solution according to the invention to this task is the general metabolic activity of bacteria which can be detected at any time in suitable nutrient media. She is u.a. most easily detectable by measuring the O ^ consumption or the COp formation per unit time.
Die erfindungsgeiaäße'Lösung selbst geht von der Tatsache aus, daß alle, eine Bakterienpopulation schädigenden Einflüsse zwangsläufig meßbare negative Konsequenzen für die Stoffwechselaktivität haben müssen. Alle bisher entwickelten bakterio-The solution according to the invention itself is based on the fact that all influences which have a damaging effect on a bacterial population must inevitably have measurable negative consequences for the metabolic activity. All previously developed bacteriological
-3- 19593 7-3- 19593 7
statisch- oder bakterizidwirksamen Chemotherapeutika hemmen bei gegebener Empfindlichkeit der Bakterien deren Stoffwechselaktivität mit entsprechendem Rückgang des Qo-Verbrauchs bzw, der COp^-Produktion. Unter Verwendung handelsüblicher, ursprünglich jedoch hierfür nicht vorgesehener, Meßinstrumente und Elektroden ist es möglich, im Rahmen einer geeigneten Testanordnung aus dem Verhalten des pO2 bzw· P0O2 Schlüsse auf die chemotherapeutische !Empfindlichkeit der jeweils vorliegenden Erregerpopulation zu ziehen.Static or bactericidal chemotherapeutic drugs inhibit their metabolic activity with a corresponding sensitivity of the bacteria with a corresponding decrease in Qo consumption or COp ^ production. Using commercially available, but originally not intended, measuring instruments and electrodes, it is possible within the framework of a suitable test arrangement from the behavior of pO 2 or · P 0 O 2 conclusions on the chemotherapeutic! Sensitivity of each existing pathogen population to draw.
Darstellung des technologischen Vorgehens am Beispiel einer HarnwegsInfekt ion:Presentation of the technological procedure using the example of a urinary tract infection:
5 - 10 ml Urin (Einsatzmenge des Untersuchungsmaterial ist abhängig von der Geometrie der Elektrode!) werden durch kurzfristiges kräftiges Schütteln mit O2 angereichert und danach eine halbe Stunde bei 37 0G inkubiert. Besteht eine behandlungsbedürftige Bakteriurie (Keimzahl > lOvml), ist mit Ablauf der Inkubation der pO2 stark abgesunken bzw· der pOOp in gleicher Größenordnung angestiegen.5 - 10 ml of urine (amount of the test material depends on the geometry of the electrode!) Are enriched by short vigorous shaking with O 2 and then incubated for half an hour at 37 0 G. If there is a bacteriuria requiring treatment (bacterial count> 10Vml), the pO 2 has dropped sharply with the expiration of the incubation or the pOOp has increased in the same order of magnitude.
Zur sich sofort anschließenden 'Bestimmung der chemotherapeutischen JSßipfindlichkeit werden je nach Zahl der zu prüfenden Chemotherapeutika gleichvolumige Proben desselben Urins mit ausreichenden Mengen der Chemotherapeutika unter Zugabe von 10 - 20 mg z.B. eines Mono- oder Disaccharids versetzt. In jeweils einer Urinprobe wird ein Chemotherapeutikum getestet! I>ie zugefügten Chemotherapeutika-Dosen bewegen sich in der Größenordnung der im Urin erreichbaren bzw. notwendigen Konzentrationen. Die .Ansätze werden ca. 2 Std. bei 37 0C inkubiert, danach zur O^-Anreicherung kräftig geschüttelt und erneut eine halbe Std. bei 37 0G inkubiert und dann der pO2 oder pC02 gemessen, !empfindlichkeit'gegenüber einem Chemotherapeutikum liegt immer dann vor, -wenn die p0o- oder pC02-Änderung sich deutlich von der einer mitzuführenden Kontrolle ohne Chemotherapeutikumzusatz im Sinne einer -Abnahme der Stoffwechsel aktivität unterscheidet.For the immediate subsequent determination of the chemotherapeutic susceptibility, depending on the number of chemotherapeutic agents to be tested, equal-volume samples of the same urine are mixed with sufficient amounts of the chemotherapeutic agents with the addition of 10-20 mg of, for example, a mono- or disaccharide. In each urine sample a chemotherapeutic agent is tested! The dosages of chemotherapeutic agents added are on the order of the levels that can be achieved or required in the urine. The mixtures are incubated for about 2 hours at 37 0 C, then vigorously shaken to O ^ enrichment and incubated again for half an hour at 37 0 G and then the pO 2 or pC0 2 measured, 'sensitivity' to a chemotherapeutic agent is always present if the change in p0 o or pC0 2 differs markedly from that of a control to be carried out without the addition of chemotherapeutic agents in the sense of a decrease in metabolic activity.
In hintereinander ablaufenden Meßvorgängen können somit im Zeitraum von ca. 3 Std. nicht nur eine Harnwegsinfektion ausge-In consecutive measuring procedures, therefore, not only a urinary tract infection can be excluded within a period of about 3 hours.
-*- 19 593 7- * - 19 593 7
schlossen oder verifiziert, sondern zugleich die Voraussetzungen für eine optimale Chemotherapie geschaffen werden. Beim z.Zt· noch susschließlich angewandten konventionellen Vorgehen benötigt man hierzu mindestens 2-3 Tage. In gleicher Weise ist das beschriebene Verfahren auch zur Testung aller anderen im Zusammenhang mit infektiösen Prozessen isolierten Bakterien anwendbar, vorausgesetzt die Bakterien werden in ausreichender Menge in ein geeignetes flüssiges Näbxme&ium, z.B. Peptonlösung, gebracht· Die beschriebene Technologie gestattet im übrigen, derartige •Testungen weitgehend zu automatisieren und damit auch größere Serien rationell zu überprüfen.closed or verified, but at the same time the conditions for optimal chemotherapy are created. In the case of the currently used conventional procedure, this requires at least 2-3 days. Likewise, the method described is also applicable to the testing of all other bacteria isolated in connection with infectious processes, provided that the bacteria are transferred in sufficient quantity to a suitable liquid seed, e.g. Peptone solution, brought · The technology described allows moreover to automate such • tests largely and thus to check even larger series efficiently.
Claims (1)
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DD76195937A DD146769A3 (en) | 1976-11-24 | 1976-11-24 | METHOD FOR TESTING THE CHEMOTHERAPEUTIC SENSITIVITY OF BACTERIA |
| SE7713070A SE7713070L (en) | 1976-11-24 | 1977-11-21 | ELECTROCHEMICAL PROCEDURE FOR QUICK DETERMINATION OF THE CHEMOTHERAPEUTIC RESPONSIBILITY OF PATOGENA MICRO-ORGANISMS |
| DK519277A DK519277A (en) | 1976-11-24 | 1977-11-23 | ELECTROCHEMICAL PROCEDURE FOR QUICK DETERMINATION OF THE CHEMOTHERAPEUTIC SENSITIVITY OF PATHOGENIC MICRO-ORGANISMS |
| DE19772752356 DE2752356A1 (en) | 1976-11-24 | 1977-11-23 | METHOD OF TESTING THE CHEMOTHERAPEUTIC SENSITIVITY OF MICROORGANISMS |
| FR7735410A FR2372229A1 (en) | 1976-11-24 | 1977-11-24 | Microbial esp. bacterial chemotherapeutic sensitivity testing - by electrochemical measurement of change in oxygen or carbon di:oxide partial pressure |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DD76195937A DD146769A3 (en) | 1976-11-24 | 1976-11-24 | METHOD FOR TESTING THE CHEMOTHERAPEUTIC SENSITIVITY OF BACTERIA |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DD146769A3 true DD146769A3 (en) | 1981-03-04 |
Family
ID=5506402
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DD76195937A DD146769A3 (en) | 1976-11-24 | 1976-11-24 | METHOD FOR TESTING THE CHEMOTHERAPEUTIC SENSITIVITY OF BACTERIA |
Country Status (5)
| Country | Link |
|---|---|
| DD (1) | DD146769A3 (en) |
| DE (1) | DE2752356A1 (en) |
| DK (1) | DK519277A (en) |
| FR (1) | FR2372229A1 (en) |
| SE (1) | SE7713070L (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5666749A (en) * | 1979-11-02 | 1981-06-05 | Kyowa Hakko Kogyo Co Ltd | Measuring method of activity of animal/botanical tissue |
| FR2622597B1 (en) * | 1987-10-30 | 1990-02-16 | Jeulin | DEVICE FOR MEASURING CELLULAR ACTIVITY |
| AU4338489A (en) * | 1988-09-20 | 1990-04-18 | Marvin Murray | Accelerated microdilution determination of bacteria susceptibility to antibiotics |
| JP2886984B2 (en) * | 1992-05-29 | 1999-04-26 | デユコーア・エル・ピー | Measuring mold growth on amorphous substrates. |
| CN119876329A (en) * | 2025-03-28 | 2025-04-25 | 浙江师范大学 | Detection method for microorganism drug sensitivity test based on threshold time and viable bacteria colony count combined calibration curve |
-
1976
- 1976-11-24 DD DD76195937A patent/DD146769A3/en unknown
-
1977
- 1977-11-21 SE SE7713070A patent/SE7713070L/en unknown
- 1977-11-23 DE DE19772752356 patent/DE2752356A1/en not_active Withdrawn
- 1977-11-23 DK DK519277A patent/DK519277A/en unknown
- 1977-11-24 FR FR7735410A patent/FR2372229A1/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| SE7713070L (en) | 1978-05-25 |
| DE2752356A1 (en) | 1978-06-01 |
| FR2372229A1 (en) | 1978-06-23 |
| DK519277A (en) | 1978-05-25 |
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