WO1995014105A2 - Antibiogramme - Google Patents

Antibiogramme Download PDF

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Publication number
WO1995014105A2
WO1995014105A2 PCT/GB1994/002551 GB9402551W WO9514105A2 WO 1995014105 A2 WO1995014105 A2 WO 1995014105A2 GB 9402551 W GB9402551 W GB 9402551W WO 9514105 A2 WO9514105 A2 WO 9514105A2
Authority
WO
WIPO (PCT)
Prior art keywords
bacteria
sample
antibiotic
test
atp
Prior art date
Application number
PCT/GB1994/002551
Other languages
English (en)
Other versions
WO1995014105A3 (fr
Inventor
Michael Stephen Roberts
Original Assignee
Aquaculture Diagnostics Limited
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
Application filed by Aquaculture Diagnostics Limited filed Critical Aquaculture Diagnostics Limited
Publication of WO1995014105A2 publication Critical patent/WO1995014105A2/fr
Publication of WO1995014105A3 publication Critical patent/WO1995014105A3/fr

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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
    • C12Q1/18—Testing for antimicrobial activity of a material

Definitions

  • This invention relates to an antibiotic sensitivity profile and in particular to a rapid antibiotic sensitivity profile for pathogens.
  • a swab or other sample containing the pathogen is inoculated onto a series of agar plates.
  • the plates are then incubated either aerobically or anaerobically for 48 hours. Assuming a selective medium has been used, any cultures should be the presumptive pathogen.
  • one further culture is made to provide a purity plate for antisera testing and a second is inoculated onto sensitivity test agar; this is a solid medium which contains no constituents which excessively enhance or reduce the inhibitory or lethal effects of the particular antibiotics tested.
  • Onto such a sensitivity agar plate are placed several small absorbent paper discs, each impregnated with a different antibiotic. The plates are then incubated for a further 48 hours.
  • the aim of this invention is to provide an antibiotic sensitivity test which obviates or mitigates these disadvantages.
  • a test comprising the steps of capturing bacteria, introducing a selected antibiotic in a predetermined inhibitory amount to the captured bacteria in a suitable cultivation medium, incubating the bacteria under conditions conducive to normal growth of the bacteria, and testing for the presence of ATP to determine the viability of the bacteria culture.
  • the method of bacteria capture is important but not critical since a number of alternatives are available, including use of an affinity column, or a microfiltration membrane which is optionally blocked in a manner known per se (so-called "dead end filtration"), or an in-line air filter, or a cross-flow and flow-through filtration technique, again optionally using blocking buffer reagents.
  • Suitable support media such as beads, particles, fibres, films and membranes may be formed from cellulose-acetate, cellulose-nitrate, regenerated cellulose, polysulphone, polyacrylonitrile, polyamide, polyimide or the like.
  • a method of determining the antibiotic sensitivity of bacteria comprising, contacting a sample to be tested with selected supported mono-clonal antibodies for a sufficient period of time to permit binding, recovering the sample containing bound target material, mixing the said sample with nutrient broth containing an antibiotic, incubating the mixed sample and subsequently testing the sample for the presence of ATP.
  • Testing is suitably carried out by use of optical detection means, such as a luminometer or spectrometer, the sample being treated with a bioluminescent or chemiluminescent reagent system.
  • optical detection means such as a luminometer or spectrometer
  • the optical detection system would be one tunable to detect the wavelength of emission appropriate for the luminescent reagent system.
  • An affinity column which contains purified highly specific mono-clonal antibodies fixed to glass or other such support beads.
  • the mono-clonal antibodies provided are specific for one particular pathogen i.e. Staphylococcus aureus to allow any of the said bacteria in a sample to be retained in the affinity column.
  • Test fluids obtained by, for example, rinsing swabs in liquid nutrients, or obtaining body fluids directly for testing, are passed through the column an amount in the range of 1 ml - 1000 ml for up to 1 hour.
  • the captured bacteria are eluted from the column using 1 ml of a suitable liquid r.f a different pH from the column. This step takes approximately 2 minutes during which time the pH is brought to near neutral.
  • Two or more micro tubes of nutrient broth each with 100 ⁇ l of the eluted bacterial suspension are added together with known LD100, (Lethal Dose 100%) amounts of antibiotics.
  • the tubes are then incubated at the optimum temperature of the particular bacteria for 8 hours. This incubation is carried out aerobically or anaerobically depending on the growth conditions favoured by the test bacteria.
  • a rapid test is carried out to detect the presence of Adenosine Triphosphate (ATP) , in this case using a bioluminescent technique employing luciferase luciferin L7.
  • ATP Adenosine Triphosphate
  • 100 ⁇ l of the incubated nutrient both containing the concentrated bacteria and the antibiotic are inoculated with 0.02% benzalkonium chloride in order to disrupt the bacterial membranes.
  • a 10 mg/ml aqueous solution of firefly lantern extract is prepared and a 100 ⁇ l amount is dispensed into a suitable cuvette. This cuvette is placed in a luminometer in which the wavelength has been set at 560 n , and thus any background luminescence can be determined and the luminometer zeroed accordingly.
  • a test kit for performing the rapid antibiotic sensitivity test may include,
  • support means such as a membrane or fibre filter, prepared for capture of bacteria and containment means therefor;

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Toxicology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

Selon l'invention, on peut obtenir un antibiogramme destiné à des agents pathogènes par un procédé comprenant les étapes consistant à prélever des bactéries, à introduire dans ces bactéries une quantité inhibitrice prédéterminée d'un antibiotique choisi, dans un milieu de culture approprié, à incuber ces bactéries dans des conditions conduisant à une croissance bactérienne normale, puis à rechercher la présence d'adénosine-triphosphate (ATP) afin de déterminer la viabilité de la culture bactérienne.
PCT/GB1994/002551 1993-11-18 1994-11-18 Antibiogramme WO1995014105A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9323760.0 1993-11-18
GB939323760A GB9323760D0 (en) 1993-11-18 1993-11-18 Antibiotic sensitivity profile

Publications (2)

Publication Number Publication Date
WO1995014105A2 true WO1995014105A2 (fr) 1995-05-26
WO1995014105A3 WO1995014105A3 (fr) 1995-06-08

Family

ID=10745356

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1994/002551 WO1995014105A2 (fr) 1993-11-18 1994-11-18 Antibiogramme

Country Status (2)

Country Link
GB (1) GB9323760D0 (fr)
WO (1) WO1995014105A2 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2348438A (en) * 1999-03-19 2000-10-04 David Anthony Stafford Detecting microbes by bioluminescence
WO2003025208A1 (fr) * 2001-09-21 2003-03-27 The Secretary Of State For Defence Procede pour determiner la presence de bacteries resistant a des antibiotiques de lyse cellulaire
WO2004090089A1 (fr) * 2003-04-09 2004-10-21 Acolyte Biomedica Limited Systeme pour une identification de micro-organismes et un test antibacterien rapides
WO2005042778A1 (fr) * 2003-10-22 2005-05-12 Acolyte Biomedica Limited Utilisation d'acides nucleiques pour essais cliniques microbiologiques
US9562253B1 (en) 2012-11-09 2017-02-07 Point Of Care Diagnostics, Llc Distinguishing between a bacterial and non-bacterial infection at the point of care

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1982004264A1 (fr) * 1981-06-04 1982-12-09 Mattiasson Bo Gustav Procede de saisie de donnees biochimiques sur des micro- organismes
WO1992016648A1 (fr) * 1991-03-18 1992-10-01 Environmental Test Systems, Inc. Test chimioluminescent pour microorganismes
EP0563858A1 (fr) * 1992-04-01 1993-10-06 Nihon Millipore Kogyo Kabushiki Kaisha Procédé de détermination quantitative des microorganismes viables

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1982004264A1 (fr) * 1981-06-04 1982-12-09 Mattiasson Bo Gustav Procede de saisie de donnees biochimiques sur des micro- organismes
WO1992016648A1 (fr) * 1991-03-18 1992-10-01 Environmental Test Systems, Inc. Test chimioluminescent pour microorganismes
EP0563858A1 (fr) * 1992-04-01 1993-10-06 Nihon Millipore Kogyo Kabushiki Kaisha Procédé de détermination quantitative des microorganismes viables

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BERCHTOLD, M. (ED.). FORTSCHRITTE DER VETERINAERMEDIZIN, NO. 35. BERICHT DES 14. KONGRESSES DER DEUTSCHEN VETERINAERMEDIZINISCHEN GESELLSCHAFT BAD NAUHEIM 9.-11. APRIL 1981 (ADVANCES IN VETERINARY MEDICINE, NO. 35. REPORT ON THE 14TH CONVENTION OF TH pages 323 - 328 WEISS R et al 'Untersuchungen zur Pr}fung der Antibiotikaempfindlichkeit von Bakterien mittels des Biolumineszenzverfahrens' *
BIOLUMIN. CHEMILUMIN., PROC. INT. BIOLUMIN. CHEMILUMIN. SYMP., 4TH (1987), MEETING DATE 1986, 491-4. EDITOR(S): SCHOELMERICH, J. PUBLISHER: WILEY, CHICHESTER, UK. CODEN: 56LVAL, 1987 Nilsson, L. et al 'Bioluminescent assay for studies of effects of antimicrobial agents on bacteria and fungi' *
J APPL BACTERIOL 56 (1). 1984. 145-150. CODEN: JABAA4 ISSN: 0021-8847 MCWALTER P W 'Determination of Susceptibility of Staphylococcus-Aureus to Methicillin by Luciferin Luciferase Assay of Bacterial ATP.' *
ZENTRALBL VETERINAERMED REIHE B 29 (5). 1982. 359-371. CODEN: ZVRBA2 ISSN: 0514-7166 WEISS R et al 'Resistenzpr}fung von Bakterien mittels der Firefly-Biolumineszenz' *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2348438A (en) * 1999-03-19 2000-10-04 David Anthony Stafford Detecting microbes by bioluminescence
WO2003025208A1 (fr) * 2001-09-21 2003-03-27 The Secretary Of State For Defence Procede pour determiner la presence de bacteries resistant a des antibiotiques de lyse cellulaire
US7648830B2 (en) 2001-09-21 2010-01-19 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland Method for determining the presence of bacteria resistant to cell lysing antibiotics
WO2004090089A1 (fr) * 2003-04-09 2004-10-21 Acolyte Biomedica Limited Systeme pour une identification de micro-organismes et un test antibacterien rapides
WO2005042778A1 (fr) * 2003-10-22 2005-05-12 Acolyte Biomedica Limited Utilisation d'acides nucleiques pour essais cliniques microbiologiques
US9562253B1 (en) 2012-11-09 2017-02-07 Point Of Care Diagnostics, Llc Distinguishing between a bacterial and non-bacterial infection at the point of care

Also Published As

Publication number Publication date
WO1995014105A3 (fr) 1995-06-08
GB9323760D0 (en) 1994-01-05

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