GB2116709A - Detecting surface mildew - Google Patents
Detecting surface mildew Download PDFInfo
- Publication number
- GB2116709A GB2116709A GB08206873A GB8206873A GB2116709A GB 2116709 A GB2116709 A GB 2116709A GB 08206873 A GB08206873 A GB 08206873A GB 8206873 A GB8206873 A GB 8206873A GB 2116709 A GB2116709 A GB 2116709A
- Authority
- GB
- United Kingdom
- Prior art keywords
- bioluminescence
- atp
- agent
- contamination
- image
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 claims description 25
- 238000005415 bioluminescence Methods 0.000 claims description 23
- 230000029918 bioluminescence Effects 0.000 claims description 23
- 239000003795 chemical substances by application Substances 0.000 claims description 21
- 239000007788 liquid Substances 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 12
- 238000011109 contamination Methods 0.000 claims description 10
- 102000004190 Enzymes Human genes 0.000 claims description 9
- 108090000790 Enzymes Proteins 0.000 claims description 9
- 241000254158 Lampyridae Species 0.000 claims description 6
- 239000003153 chemical reaction reagent Substances 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 238000011169 microbiological contamination Methods 0.000 claims description 5
- 239000004744 fabric Substances 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 4
- 239000004753 textile Substances 0.000 claims description 4
- 238000011005 laboratory method Methods 0.000 claims description 2
- 238000010998 test method Methods 0.000 claims description 2
- 238000012800 visualization Methods 0.000 claims description 2
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 27
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 27
- 210000004027 cell Anatomy 0.000 description 13
- 239000005082 bioluminescent agent Substances 0.000 description 5
- 238000004020 luminiscence type Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
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
-
- 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
- G01N33/36—Textiles
- G01N33/367—Fabric or woven textiles
-
- 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
- C12Q2304/00—Chemical means of detecting microorganisms
- C12Q2304/60—Chemiluminescent detection using ATP-luciferin-luciferase system
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Textile Engineering (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Food Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Toxicology (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A method has been developed for detecting the presence and even the location of mildew on textile fabrics, and by extension the presence of any living organism at the surface of any object, by the application of an agent for the release, from the cells of the organism, of a substance, exemplified by adenosine triphosphate, which is capable of reaction with a specific enzyme to produce bioluminescence. The bioluminescence is visualised by an image intensifier. The released substance may be allowed to migrate from the surface and be quantitatively determined, or it may be visualised in situ to show the location of the contamination.
Description
SPECIFICATION
Microbiological testing
The invention relates to methods of testing for the presence of living organisms at the surface of an object.
It has been known for several years that all living organisms contain substances contain adenosine triphosphate (ATP), and that hydrolytic reaction of
ATP with certain enzymes results in the production of light, although the mechanism of the reaction is not necessarily fully understood. The amount of light, referred to as bioluminescence, emitted is very small, and it has been the practice, in laboratory experiments with the reaction, to determine the light emission by use of a photometer.
The ATP is contained in cells within the organisms, and can be released from the cells by the application of a reagent capable of attacking the cell membrane.
It has now been realised that the reaction can be adapted to provide a simple, rapid and essentially non-destructive test for the contamination by microorganisms of the surface of objects such as pieces of textile fabric.
ATP is, in fact, only one of a group of biochemical substances, each of which is capable of reaction with a specific enzyme to produce bioluminescence, and although in the following specification reference will be made for the sake of simplicity primarily to ATP, it should be understood that the term is to be contrued as extending to any member of the aforementioned group or a derivative thereof.
According to the invention there is provided a method of test for the presence of living organisms having ATP-bearing cells at the surface of an object, comprising applying to the surface an agent for the release of ATP from the cells, providing a bioluminescent agent for reaction with released ATP to produce bioluminescence, and observing the luminescence produced.
The releasing agent may be in solution or suspension in a liquid, and may be applied by spraying or may be poured onto the surface. The said releasing agent may be applied to the surface first whereby the ATP is released into the liquid and the bioluminescent agent is then added to the liquid.
Alternatively, the bioluminescent agent, in solution or suspension, may be applied to the surface first and the releasing agent is then applied to the thus moistened surface.
The observation may be made by the use of an image intensifier.
Embodiments of the invention will now be described by way of example, with reference to two techniquesforthevisualisation of microbiological contamination of the surface of a piece of textile fabric.
In the first example, a solution of NRB nucleotide releasing reagent for microbial cells is poured onto the surface of the sample in a shallow dish. The reagent has the effect of disrupting the membranes of the cells in any living organism present on or near the surface, such organism being, for example, that responsible for the defect commonly known as mildew. The disruption of the cell membranes allows ATP, which is known to occur in living organisms, to be released from within the cells and to enter the solution so as to spread over substantially the whole area of the dish.
A bioluminescent agent, Firefly luciferinluciferase, such as Limit HS (trade mark), is rehydrated with a buffer to give a solution of pH 7.75, a chelating agent is then added, and the solution is added, at a temperature in the range 20-25 degrees
C, to the liquid in the dish.
The effect of the bioluminescent agent is to react with released ATP in the liquid so as to produce light in quantity proportional to the quantity of ATP released from the sample. It is understood that only about one photon of light is produced from a single molecule of ATP, and unless the sample had been heavily contaminated, and thus a large quantity of
ATP had been released, the light emitted would be hardly visible to the naked eye though possibly sufficient to affect a sensitive photographic film.
However, even in the case of relatively slight contamination, the luminescence of the liquid in the dish can be visualised by the use of an image intensifier, such as Mullard (trade mark) XX1500TV which has a gain of approximately 65,000. By the use of the intensifier the luminescence provided by the sample can thus be directly observed by the eye and a semi-quantitative estimate can be made of the contamination present relative to that present in a standard or another unknown sample. A permanent record of the intensified image of the luminescent liquid can be produced by use of a camera or video recorder.
In a particularly advantageous application of the method, a visual image showing the precise location of contamination at a surface is provided. The method is particularly suitable for the examination of surfaces in situ and may be used to follow variations in contamination with time.
The method involves first applying the Firefly luciferin-luciferase enzyme, prepared as mentioned above, onto the surface to be investigated, conveniently by spraying. No significant reaction with ATP from surface contamination occurs at this stage because the ATP is predominantly enclosed within cell membranes. Commercially available firefly enzyme may itself be slightly contaminated with ATP however and there may be a slight bioluminescent background from reaction of the enzyme with such traces of ATP, and such background luminescence should be discounted in considering that revealed after the next subsequent step.
NRB nucleotide releasing reagent is then sprayed onto the surface wetted with the enzyme solution. As the ATP-bearing cells of any micro organisms present at the surface of the sample are disrupted by the releasing agent, the ATP is released into an environment of firefly enzyme whereby bioluminescence is emitted in the immediate vicinity of the cell from which the ATP was released.
If sufficient quantities of liquid were added, the bioluminescence would eventually spread over the whole wetted area, but in the presence of minimum liquid and over a relatively short period of time, the bioluminescence provides a substantially accurate image of the location and concentration of ATPbearing cells and thus of microbiological contamination.
By use of an image intensifier, the image can be seen by the eye or recorded by a camera. The image intensifier may be provided with a video camera and recording system, and provided that the reagents applied do not inhibit the growth of the microorganism, the technique would appear to provide a method for observing and recording the developing extent of contamination as a function of time.
A particularly advantageous aspect of the invention lies in the fact that the sample can be examined without any handling, which would involve the risk of itself adding undesirable microbilogical contamination, and without resorting to masseration as employed in the laboratory techniques hitherto employed for demonstrating bioluminescence.
Whilst in the forgoing description reference has been made to the use only of Firefly enzyme to act as a bioluminescence agent with the ATP occuring in living organisms to produce bioluminescence, methods are to be regarded as being within the scope of the invention which employ other agents which result in the emission of bioluminescence on reaction with biochemical molecules or derivatives thereof.
CLAIMS (Filed on 3 March 83)
1. A method oftesting forthe presence, at the surface of an object, of living organisms having
ATP-bearing cells, comprising applying to said surface an agent for releasing ATP from the cells, providing agent for reaction with released ATP to produce bioluminescence, and observing the bioluminescence produced.
2. A method according to Claim 1 wherein the releasing agent is applied to the surface in solution or suspension in a liquid.
3. A method according to Claim 2 wherein the bioluminescence agent is subsequently added to the liquid.
4. A method according to Claim 1 wherein the bioluminescence agent in solution or suspension is added to the surface first and the releasing agent is then applied to the thus moistened surface.
5. A method according to any one of the preceding claims wherein the observation is made by use of an image intensifier.
6. A method according to Claim 5 wherein the image produced by the intensifier is recorded.
7. A method according to any one of the preceding claims used for visualisation of microbiological contamination of a textile fabric.
8. A method of testing for the presence, at the surface of an object, of living organisms having
ATP-bearing cells, substantially as described.
**WARNING** end of DESC field may overlap start of CLMS **.
Claims (8)
1. A method oftesting forthe presence, at the surface of an object, of living organisms having
ATP-bearing cells, comprising applying to said surface an agent for releasing ATP from the cells, providing agent for reaction with released ATP to produce bioluminescence, and observing the bioluminescence produced.
2. A method according to Claim 1 wherein the releasing agent is applied to the surface in solution or suspension in a liquid.
3. A method according to Claim 2 wherein the bioluminescence agent is subsequently added to the liquid.
4. A method according to Claim 1 wherein the bioluminescence agent in solution or suspension is added to the surface first and the releasing agent is then applied to the thus moistened surface.
5. A method according to any one of the preceding claims wherein the observation is made by use of an image intensifier.
6. A method according to Claim 5 wherein the image produced by the intensifier is recorded.
7. A method according to any one of the preceding claims used for visualisation of microbiological contamination of a textile fabric.
8. A method of testing for the presence, at the surface of an object, of living organisms having
ATP-bearing cells, substantially as described.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB08206873A GB2116709B (en) | 1982-03-09 | 1982-03-09 | Microbiological testing detecting surface mildew |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB08206873A GB2116709B (en) | 1982-03-09 | 1982-03-09 | Microbiological testing detecting surface mildew |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| GB2116709A true GB2116709A (en) | 1983-09-28 |
| GB2116709B GB2116709B (en) | 1986-02-19 |
Family
ID=10528883
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| GB08206873A Expired GB2116709B (en) | 1982-03-09 | 1982-03-09 | Microbiological testing detecting surface mildew |
Country Status (1)
| Country | Link |
|---|---|
| GB (1) | GB2116709B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1986007094A1 (en) * | 1985-05-22 | 1986-12-04 | University Of Wales College Of Medicine | Bacterial cell extractant |
| WO1987006707A1 (en) * | 1986-04-29 | 1987-11-05 | Murex Corporation | Bound assay components |
| GB2246197A (en) * | 1990-07-12 | 1992-01-22 | Bio Rad Laboratories | Detection and imaging in biochemical assays using phosphorescent screens |
| GB2294115A (en) * | 1994-10-11 | 1996-04-17 | Whitlock Hughes Ltd | Measuring surface contamination by organisms |
| GB2348438A (en) * | 1999-03-19 | 2000-10-04 | David Anthony Stafford | Detecting microbes by bioluminescence |
| GB2358124A (en) * | 2000-01-14 | 2001-07-18 | Chemisphere Uk Ltd | Method of cleaning and monitoring in ware washing machines |
| JP2002525123A (en) * | 1998-09-30 | 2002-08-13 | パッカード バイオサイエンス ベスローテン フェンノートシャップ | How to detect ATP |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1379506A (en) * | 1971-05-25 | 1975-01-02 | Baxter Laboratories Inc | Receptacle having a distendable sidewall |
| GB1550076A (en) * | 1976-09-30 | 1979-08-08 | Battelle Institut E V | Methods of and samplers suitable for taking and collecting sample material |
| GB2073885A (en) * | 1980-04-15 | 1981-10-21 | Whitlock G D | Method of and apparatus for detecting the presence of live organisms in substances |
| GB1601031A (en) * | 1977-01-31 | 1981-10-21 | Plakas C J | Method for detection of bacterial concentration by luminescence |
| GB1604249A (en) * | 1977-05-31 | 1981-12-02 | Kolehmainen S | Selective measurement of somatic and microbial cells |
-
1982
- 1982-03-09 GB GB08206873A patent/GB2116709B/en not_active Expired
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1379506A (en) * | 1971-05-25 | 1975-01-02 | Baxter Laboratories Inc | Receptacle having a distendable sidewall |
| GB1550076A (en) * | 1976-09-30 | 1979-08-08 | Battelle Institut E V | Methods of and samplers suitable for taking and collecting sample material |
| GB1601031A (en) * | 1977-01-31 | 1981-10-21 | Plakas C J | Method for detection of bacterial concentration by luminescence |
| GB1604249A (en) * | 1977-05-31 | 1981-12-02 | Kolehmainen S | Selective measurement of somatic and microbial cells |
| GB2073885A (en) * | 1980-04-15 | 1981-10-21 | Whitlock G D | Method of and apparatus for detecting the presence of live organisms in substances |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1986007094A1 (en) * | 1985-05-22 | 1986-12-04 | University Of Wales College Of Medicine | Bacterial cell extractant |
| WO1987006707A1 (en) * | 1986-04-29 | 1987-11-05 | Murex Corporation | Bound assay components |
| GB2246197A (en) * | 1990-07-12 | 1992-01-22 | Bio Rad Laboratories | Detection and imaging in biochemical assays using phosphorescent screens |
| GB2246197B (en) * | 1990-07-12 | 1994-03-16 | Bio Rad Laboratories | Detection and imaging in biochemical assays using phosphor screens |
| GB2294115A (en) * | 1994-10-11 | 1996-04-17 | Whitlock Hughes Ltd | Measuring surface contamination by organisms |
| JP2002525123A (en) * | 1998-09-30 | 2002-08-13 | パッカード バイオサイエンス ベスローテン フェンノートシャップ | How to detect ATP |
| GB2348438A (en) * | 1999-03-19 | 2000-10-04 | David Anthony Stafford | Detecting microbes by bioluminescence |
| GB2358124A (en) * | 2000-01-14 | 2001-07-18 | Chemisphere Uk Ltd | Method of cleaning and monitoring in ware washing machines |
| GB2358124B (en) * | 2000-01-14 | 2003-08-20 | Chemisphere Uk Ltd | Method of cleaning and monitoring in ware washing machines |
Also Published As
| Publication number | Publication date |
|---|---|
| GB2116709B (en) | 1986-02-19 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 732 | Registration of transactions, instruments or events in the register (sect. 32/1977) | ||
| PCNP | Patent ceased through non-payment of renewal fee |
Effective date: 19980309 |