Processing

Please wait...

PATENTSCOPE will be unavailable a few hours for maintenance reason on Saturday 31.10.2020 at 7:00 AM CET
Settings

Settings

Goto Application

1. WO1996002666 - CAPTURE ASSAYS

Publication Number WO/1996/002666
Publication Date 01.02.1996
International Application No. PCT/GB1995/001643
International Filing Date 12.07.1995
Chapter 2 Demand Filed 08.02.1996
IPC
C12Q 1/04 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
02involving viable microorganisms
04Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
C12Q 1/48 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
48involving transferase
G01N 33/569 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
569for microorganisms, e.g. protozoa, bacteria, viruses
CPC
C12Q 1/04
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
02involving viable microorganisms
04Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
C12Q 1/48
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
48involving transferase
G01N 33/569
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
569for microorganisms, e.g. protozoa, bacteria, viruses
Applicants
  • THE SECRETARY OF STATE FOR DEFENCE [GB]/[GB] (AllExceptUS)
  • SQUIRRELL, David, James [GB]/[GB] (UsOnly)
Inventors
  • SQUIRRELL, David, James
Agents
  • BECKHAM, Robert, William
Priority Data
9414096.913.07.1994GB
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) CAPTURE ASSAYS
(FR) DOSAGES PAR DETECTION
Abstract
(EN)
A method is provided for determining the presence and/or amount of a microorganism and/or its intracellular material present in a sample characterised in that:(a) exposing the sample to a specific binding agent that has been immobilised upon a solid substrate, the specific binding agent being capable of binding to the microorganism or its intracellular material such that it becomes associated with the solid substrate, (b) exposing the solid substrate to an agent capable of making adenylate kinase associated with the microorganism and/or its intracellular material accessible to solutions applied to the substrate, (c) applying a solution containing adenosine diphosphate (ADP) to the substrate under conditions whereby adenosine triphosphate (ATP) may be produced by any adenylate kinase present, and (d) measuring the amount of adenosine triphosphate (ATP) and relating that to the presence and/or amount of microorganism or intracellular contents. Step (d) may be carried out using an assay that includes a colour forming reaction, but is preferably carried out by use of luciferase/luciferin reagent to produce light proportional to the amount of ATP produced, and that is detected using a luminometer. Preferably step (c) includes presence of magnesium ions at a molar concentration sufficient to allow maximal conversion of ADP to ATP. Most preferably step (b) and (c) are carried out by adding extractant, ADP and magnesium ions to the sample and incubating the mixture for a predetermined period to effect conversion of ADP to ATP.
(FR)
On décrit un procédé destiné à déterminer dans un échantillon la présence et/ou le niveau d'un micro-organisme et/ou la substance intracellulaire présente dans celui-ci, ledit procédé consistant: (a) à exposer l'échantillon à un agent de liaison spécifique qui a été immobilisé sur un substrat solide, cet agent de liaison étant susceptible de se lier au micro-organisme ou à la substance intracellulaire de celui-ci et s'associe au substrat solide; (b) à exposer le substrat solide à un agent susceptible d'induire l'association de l'adénylate-kinase au micro-organisme et/ou à la substance cellulaire de celui-ci accessible aux solutions appliquées sur le substrat; (c) à appliquer une solution contenant de l'adénosine-diphosphate (ADP) sur le substrat dans des conditions dans lesquelles l'adénosine-triphosphate (ATP) peut être produite par une quelconque adénylate-kinase présente, et (d) à mesurer le niveau d'adénosine-triphosphate (ATP) et à établir un rapport entre ce niveau et la présence et/ou les niveaux du micro-organisme ou de la teneur intracellulaire de celui-ci. On peut effectuer l'étape (d) en utilisant un dosage comprenant une réaction de formation de couleur, mais on préfère effectuer celle-ci en utilisant un réactif de type luciférase/luciférine afin de produire une lumière proportionnelle au niveau d'ATP produite, cette lumière étant détectée à l'aide d'un luminomètre. De préférence, l'étape (c) comprend la présence d'ions magnésium à une concentration molaire suffisante pour permettre une conversion maximale d'ADP en ATP. Les étapes (b) et (c) sont préférablement réalisées par l'addition dans l'échantillon d'un solvant d'extraction, de l'ADP ainsi que des ions magnésium et par mise en incubation du mélange pendant une période de temps prédéterminée pour induire la conversion d'ADP en ATP.
Latest bibliographic data on file with the International Bureau