US20100105093A1 - Assay method for the detection of viable microbial cells in a sample - Google Patents

Assay method for the detection of viable microbial cells in a sample Download PDF

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US20100105093A1
US20100105093A1 US12/375,724 US37572407A US2010105093A1 US 20100105093 A1 US20100105093 A1 US 20100105093A1 US 37572407 A US37572407 A US 37572407A US 2010105093 A1 US2010105093 A1 US 2010105093A1
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atp
sample
assay
assay method
phosphate
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Gerald J. Bugler
Catherine M. Ramsay
William J. Simpson
Mark B. Driscoll
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3M Innovative Properties Co
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Assigned to 3M INNOVATIVE PROPERTIES COMPANY reassignment 3M INNOVATIVE PROPERTIES COMPANY ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: SIIMPSON, WILLIAM J., BUGLER, GERALD J., DRISCOLL, MARK B., RAMSAY, CATHERINE M.
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    • 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
    • C12Q1/66—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving luciferase
    • 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
    • C12Q1/06—Quantitative determination

Definitions

  • Microbial contamination of samples may be due to a number of different types of microorganisms.
  • the microorganisms may be present as either single colonies or multiple types of microbe colonies and for example, bacteria and fungi may be present in equal or differing amounts in or on the sample to be tested.
  • ATP Adenosine triphosphate
  • the firefly luciferase reaction for ATP employs a purified enzyme (firefly luciferase), a substrate (D-luciferin), magnesium ions and oxygen.
  • a purified enzyme firefly luciferase
  • D-luciferin a substrate
  • magnesium ions magnesium ions
  • Light produced through the reaction of ATP with D-luciferin and oxygen is measured with a luminometer.
  • the intensity of light produced by the reaction is proportional to the amount of ATP present in the sample.
  • the light produced in the reaction is emitted as a continuous glow which makes the assay more reliable, as it reduces the reliance on precise timing to view a result on the part of the analyst.
  • ATP degrading enzymes are known to be used to remove extracellular ATP prior to extraction of cellular ATP. Suitable enzymes used include ecto-ATPases among others and specifically apyrases. Apyrases are enzymes that catalyse the hydrolysis of ATP, producing adenosine diphosphate (ADP) and inorganic phosphate. In a second step they can further degrade ADP to adenosine monophosphate (AMP) and inorganic phosphate. For optimum activity, they require divalent cations, such as calcium or magnesium.
  • Apyrases can be obtained from a range of sources including potatoes, peas, and bacteria. Apyrases from a variety of sources display a large degree of homology, particularly in relation to the active site of the enzyme.
  • the removal of ATP by apyrase is not a rapid process. Typically it is the slowest step in the entire biomass estimation procedure which is not desirable with high throughput sampling. While the extraction and ATP measurement steps can be carried out within a timeframe of seconds, removal of extracellular ATP using apyrase requires a timescale of minutes. Treatment periods in the range 5-30 minutes are typical.
  • Assay protocols have been devised in which apyrase is immobilized on a solid support to provide a localized concentration of enzyme which allows the contact time between apyrase and extracellular ATP to be reduced. Even so, several minutes are still required for removal of ATP. Further, increasing the concentration of enzyme can contribute to an additional problem, in that there may be residual apyrase activity during the extraction step. This leads to sample instability, making the overall assay both time-sensitive and operator-dependent.
  • a complementary strategy may be to use additional enzymes to assist in destruction of ATP by apyrase. Such enzymes can include adenosine phosphate deaminase, hexokinase and adenosine triphosphatase. However, this additional step can increase assay times, and increase the cost of performing the assay as additional reagents are used.
  • a second problem associated with known techniques is that the ability of apyrase to remove ATP from test samples is often less than that seen in the case of model solutions. This is due to the presence of materials in test samples which inhibit the activity of apyrase.
  • ortho-vanadate has been used to inhibit apyrase activity when measuring concentrations of extracellular ATP remaining after apyrase treatment of beer samples and prior to ATP extraction.
  • ortho-vanadate has drawbacks in that it is highly toxic to both users and the microbial cells that are being assayed, so false readings can result.
  • Gadolinium has been shown to inhibit apyrase activity. However, this element has antimicrobial properties and is thus unsuitable for inhibiting potato apyrase in microbiological tests.
  • ATP assays to inhibit apyrase activity prior to ATP extraction
  • compounds which have been considered and rejected for use in ATP assays to inhibit apyrase activity prior to ATP extraction include but are not limited to: sodium deoxycholate, sodium fluoride, sodium azide, and various other small molecule inhibitors. While some of these materials are relatively good inhibitors of apyrase, all suffer from drawbacks with respect to their use in tests for live microorganisms. Some are inhibitory to microbial cell metabolism; some are hazardous to users; while others strongly inhibit the activity of firefly luciferase. Proteolytic enzymes, including trypsin, chymotrypsin, subtilisin, proteinase K and papain have also been used.
  • the invention relates to an assay method for the detection of viable microbial cells in a sample.
  • the invention relates to the assessment of the cleanliness of surfaces and the accurate measurement of the numbers of live microorganisms (viable microbial cells/viable cell biomass and which forms are interchangeable) in various types of samples including solids, liquids and gases.
  • live microorganisms viable microbial cells/viable cell biomass and which forms are interchangeable
  • microorganism includes bacteria, yeasts, fungi or algal blooms.
  • the present invention provides an assay with apyrase inhibitors that:
  • the present invention provides a quick, efficient and accurate indication of the level of viable microbial cells in a sample.
  • phosphorous-containing materials inhibit apyrase activity. Inhibition of apyrase activity by the phosphorous/phosphate containing materials is unexpected because assays involving the use of apyrase have traditionally used the detection of phosphate production as the measure of activity of the enzyme. This has essentially precluded the use of phosphorous-containing buffers or compounds in the test solution in order to avoid assay interference.
  • an assay method for the detection of viable microbial cells in a sample comprising the steps of:
  • the method includes the step of first isolating a sample suspected of containing viable microbial cells having intracellular ATP and extraneous material having extracellular ATP.
  • an extractant is added to the sample prior to assaying the level of undegraded ATP in order to extract undegraded ATP from the sample.
  • the ATP degrading enzyme is an ectoATPase.
  • the ectoATPase is an apyrase.
  • the ATP degrading enzyme such as the apyrase is buffered.
  • the apyrase reagent is buffered to have a pH value of substantially 6.5.
  • Standard buffers may be used such as HEPES Buffer.
  • the pH may be in the range of substantially 9 to 5, 8 to 6 or 7.5 to 6.2.
  • the apyrase is of a concentration in the range of 1 ml apyrase to 10 ⁇ of 10 ⁇ 7 M of ATP.
  • the apyrase reaction is typically carried out at room temperature and a typical reaction time is 1-30 minutes, more typically 15 minutes.
  • divalent cations are added to a reagent including the apyrase. It is preferred that the divalent cations are calcium ions and/or magnesium ions. The addition of divalent cations is beneficial because apyrase has low activity in the presence of substances which can bind to divalent cations since it requires calcium ions (or other divalent cations) for maximum activity. Typically a solution of Magnesium Chloride is used at a typical concentration of 5 mM.
  • the phosphate containing compound is selected from one or more of sodium phosphate, potassium phosphate, sodium pyrophosphate, penta-sodium triphosphate or sodium polyphosphate.
  • the extractant is a surface-active agent that is used to extract ATP from live cells.
  • the surface-active agents include one or more of quaternary ammonium compounds, dialkyl ammonium salts and bis-biguanides.
  • the extractant is selected from one or more of the group consisting of N—N-dimethyldodecan-1-amine (GENAMIN), chloroprozamine, liquid lysate (LL1), dichlorogallium (III) ⁇ -diketonato derivatives GaCl 2 (BDK) and combinations thereof.
  • GAAMIN N—N-dimethyldodecan-1-amine
  • chloroprozamine chloroprozamine
  • LL1 liquid lysate
  • BDK dichlorogallium
  • BDK dichlorogallium
  • the extractants are used at a concentration of 0.05-500 and more typically 0.1-5 g/l.
  • the extractant period is carried out 3-10 minutes, more typically 4-8 minutes and most preferably for a period of substantially 5 minutes.
  • the ATP is extracted at room temperature, which reduces the risk of viable cells being killed by elevated temperatures.
  • one or more neutralizing agents are added to the sample containing the extractant. This has the benefit, in particular with surface-active agents, of inactivating the reagents so they do not have a destructive effect on the viable microorganisms in the sample.
  • the assay is a light emitting assay.
  • light emitting assay to monitor the levels of ATP is a luciferase assay.
  • phosphate-containing materials in the sample are precipitated prior to performing the apyrase reaction.
  • the residual activity of apyrase in assays for viable microbial cells is inhibited by incorporating the one or more phosphate-containing substances in a reagent which is added to the sample matrix after the apyrase treatment step.
  • a second reagent is added to the sample.
  • a second reagent is added that is designed to increase intracellular ATP levels by manipulation of cellular metabolism, an ATP extraction reagent, or an ATP detection reagent containing luciferase and luciferin. These are illustrative of the materials that can be used and are not intended to be limiting.
  • an assay kit for the detection of viable microbial cells in a sample including a container including an ATP degrading enzyme, a container with a phosphate containing compound together with means to combine the enzyme and phosphate containing compound to carry out an assay method comprising the steps of:
  • the assay kit includes an extractant to be added to the sample prior to assaying the level of undegraded ATP to extract the undegraded ATP from the sample.
  • the ATP degrading enzyme is an ectoATPase.
  • the ectoATPase is an apyrase.
  • the ATP degrading enzyme such as the apyrase is provided as a buffered solution at a pH value repeat range of substantially 6.5.
  • the apyrase is of a concentration in the range of 1 ml apyrase to 10 ⁇ l of 10 ⁇ 7 M of ATP. A typical value is 0.96 units per ml.
  • an appropriate amount of apyrase can be adjusted for a particular reaction.
  • divalent cations are included in a reagent including the apyrase.
  • the divalent cations are calcium ions and/or magnesium ions at a typical concentration of 5 mM.
  • the phosphate containing compound is selected from one or more of sodium phosphate, potassium phosphate, sodium pyrophosphate, penta-sodium triphosphate or sodium polyphosphate.
  • the extractant can be a surface-active agent that is used to extract ATP from live cells. It is preferred that the surface-active agents include one or more of quaternary ammonium compounds, dialkyl ammonium salts and bis-biguanides. More preferably, the extractant is selected from one or more or the group consisting for example of N—N-dimethyldodecan-1-amine (GENAMIN), chloroprozamine, liquid lysate (LL1), dichlorogallium(III) ⁇ -diketonato derivatives GaCl 2 (BDK).
  • GAAMIN N—N-dimethyldodecan-1-amine
  • LL1 liquid lysate
  • BDK dichlorogallium(III) ⁇ -diketonato derivatives GaCl 2
  • the extractants are used at a concentration of 0.05-50 g/l more typically 0.1-5 g/l.
  • the assay kit includes neutralizing agents which are added to the sample containing the extractant.
  • FIGS. 1.1 , 2 . 1 , 3 . 1 and 4 . 1 show the Relative Light Units (RLU) provided by samples using different extractants, one sample having had a phosphate compound added, and the other sample being a control in which phosphate has not been added;
  • RLU Relative Light Units
  • FIGS. 1.2 , 2 . 2 , 3 . 2 and 4 . 2 show the amount of ATP released from samples using different extractants both with and without phosphate included after 5 and 10 minutes;
  • FIGS. 1.3 , 2 . 3 , 3 . 3 and 4 . 3 show the light emitted by samples containing various extractants with and without phosphate over a period of 70-80 seconds.
  • Leuconostoc sp MD110 was grown by streaking a bead onto Tryptone Soya Broth agar to revive bead for stock (place in 30° C. incubator overnight)
  • Cells for the experiment were prepared using a 100 ⁇ dilution of this stock with a sterile solution of glucose (10 g/l) and MgCl 2 (5 mM) at room temperature. Cells were diluted into this solution at least 20 minutes prior to starting each experiment to allow adaptation to the new conditions.
  • the glucose provides energy for the cells, and the magnesium salt facilitates apyrase activity in the subsequent assays.
  • luciferase enzyme was added to one of the cuvette (a luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo) reconstituted with 17.5 ml of HEPES buffer) and read in a luminometer.
  • the cuvette was removed from the luminometer and 10 ⁇ l of 10 ⁇ 7 M ATP was added to standardize with ATP.
  • steps 4-10 were repeated using a solution of long chain fatty acid ethoxylates (1 g/l) without phosphate.
  • FIG. 1.1 shows the percentage RLU remaining after 5 minutes extraction with each extractant.
  • the assay that had 30 mM phosphate mixed with the extractant experienced a 38% rise in RLU from T-5 to T-10.
  • the assay that contained no phosphate there was a 9% reduction in RLU from T-5 to T-10.
  • the increase in RLU seen in the assay containing phosphate may be explained by the slow extraction time of a solution of long chain fatty acid ethoxylates. More ATP was being extracted over time, whilst the action of apyrase was reduced due to the presence of phosphate.
  • Leuconostoc sp MD110 was grown by streaking a bead onto de Man Rogosa Sharpe agar to revive bead for stock (place in 30° C. incubator overnight).
  • Cuvette 1 was read in a luminometer after 5 minutes extraction time.
  • the sample was returned to the luminometer and read again, then read 5 consecutive times in a row in order to determine the final kinetics of the reaction.
  • the kinetics of the two assays ( FIG. 2.3 ) provide clear evidence of phosphate inhibition of apyrase.
  • the control assay signal drops by a factor of 10 in 50 seconds, whilst the assay containing phosphate drops only 10% in this time.
  • Leuconostoc sp MD110 was grown by streaking a bead onto de Man Rogosa Sharpe agar to revive bead for stock (place in 30° C. incubator overnight).
  • Cells for the experiment were prepared using a 100 ⁇ dilution of this stock with a sterile solution of glucose (10 g/l) and MgCl 2 (5 mM) at room temperature. Cells were diluted into this solution at least 20 minutes prior to starting each experiment to allow adaptation to the new conditions.
  • a special luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo) enzyme was prepared. This was reconstituted in 17.5 ml of 30 mM phosphate/10 g/l a solution of long chain fatty acid ethoxylates.
  • the sample was returned to the luminometer and read again, then read 5 consecutive times in a row in order to determine the final kinetics of the reaction.
  • Steps 7-11 were repeated for cuvette 2 .
  • steps 4 to 13 were repeated but using a luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo) made by reconstituting in 25 mM HEPES buffer pH 7.75 with a solution of long chain fatty acid ethoxylates (10 g/l).
  • a detergent-resistant luciferase Promega, Ultraglo
  • the kinetics of the luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo)/phosphate/solution of long chain fatty acid ethoxylates assay are much more stable than the kinetics of the luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo)/HEPES/solution of long chain fatty acid ethoxylates assay ( FIG. 3 ).
  • the RLU values for the luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo)/HEPES/solution of long chain fatty acid ethoxylates assay dropped by 97.4% in 75 seconds, whilst the luciferase/luciferin reagent formulated with a detergent-resistant luciferase (Promega, Ultraglo)/phosphate/solution of long chain fatty acid ethoxylates assay RLU values increased by 13% in this time.
  • Leuconostoc sp MD110 was grown by streaking a bead onto de Man Rogosa Sharpe agar to revive bead for stock (place in 30° C. incubator overnight).
  • Cells for the experiment were prepared using a 100 ⁇ dilution of this stock with a sterile solution of glucose (10 g/l) and MgCl 2 (5 mM) at room temperature. Cells were diluted into this solution at least 20 minutes prior to starting each experiment to allow adaptation to the new conditions.
  • the suspension was then treated with 1 ml of reconstituted apyrase 0.96 units per ml, and 10 ⁇ l of ATP (10 ⁇ 7 M) was added and left for 15 minutes at room temperature.
  • the sample was returned to the luminometer and read again, then read 5 consecutive times in a row in order to determine the final kinetics of the reaction.
  • Steps 7-11 were repeated for cuvette 2 .
  • steps 4 to 13 were repeated but using 20 ⁇ l of Reverse Osmosis water in place of 20 ⁇ l phosphate.
  • the Kinetics for the assays again indicate that phosphate inhibition of apyrase.
  • the control assay RLUs drop by around 12,000 RLU in 46 seconds, whilst the RLU values of the assay containing phosphate drop by around 1,400 RLU in this period.
  • the graphs shown indicate that phosphate inhibits the action of apyrase. This has the surprising technical effect of providing a method that can be used for analysing the levels of viable cell biomass. After removal of extracellular ATP, the reaction is stopped by the phosphate containing material so that ATP levels which are present due to viable cell biomass can then be used to measure the levels of microorganisms in a sample. There is a stage of removal of extracellular material followed by a stage in which this removal activity is halted so a more accurate indication of the levels of viable cell biomass can be achieved.

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130338350A1 (en) * 2012-06-15 2013-12-19 Ut-Battelle, Llc Method for isolating nucleic acids
US8993260B2 (en) 2012-05-02 2015-03-31 Charles River Laboratories, Inc. Fluorescence-based viability staining method using a membrane permeable flourescent dye and membrane impermeable fluorescence quencher
US9709500B2 (en) 2012-05-02 2017-07-18 Charles River Laboratories, Inc. Optical method for detecting viable microorganisms in a cell sample
JP2017533718A (ja) * 2014-11-13 2017-11-16 スリーエム イノベイティブ プロパティズ カンパニー 試料中の細菌ａｔｐを検出するためのａｔｐ−ジホスホヒドロラーゼを含むキット
US10324036B2 (en) 2012-05-02 2019-06-18 Charles River Laboratories, Inc. Porous planar cell capture system

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5177524B2 (ja) * 2008-07-02 2013-04-03 キッコーマン株式会社 茶系飲料中微生物の測定方法
CA2946721C (en) * 2014-04-24 2022-10-25 Point Of Care Diagnostics, Llc Rapid determination of bacterial susceptibility to an antibiotic at the point of care
ES2791302T3 (es) * 2014-11-07 2020-11-03 Apirays Bioscience Ab Métodos analíticos y de diagnóstico que utilizan apirasa de Shigella flexneri
EP3184644A1 (de) * 2015-12-22 2017-06-28 Omya International AG Lebensfähigkeitsprüfung mikrobieller zellen zur detektion oder bestimmung von schlammkontamination
EP4308696A1 (de) 2021-03-16 2024-01-24 ApiRays Bioscience AB Modifizierte shigella-apyrase und verwendungen davon
CN113249428A (zh) * 2021-05-17 2021-08-13 厦门承葛医学检验实验室有限公司 一种快速检测活菌量的方法
CN118215409A (zh) * 2021-06-29 2024-06-18 国际营养与健康丹麦私人有限公司 细胞培养方法
US12350388B2 (en) 2021-09-10 2025-07-08 B/E Aerospace, Inc. Methods for measuring and quantifying surface cleanability of articles
WO2023196503A1 (en) 2022-04-06 2023-10-12 Promega Corporation Atp detection

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3408440A (en) * 1966-09-26 1968-10-29 Procter & Gamble Method of preparing substantially cell-free somatic antigens
US3745090A (en) * 1970-08-04 1973-07-10 Nasa Method of detecting and counting bacteria in body fluids
US5004684A (en) * 1987-09-22 1991-04-02 Lumac Bv Method for ATP extraction

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4014745A (en) * 1975-04-30 1977-03-29 Nasa Application of luciferase assay for ATP to antimicrobial drug susceptibility
GB1604249A (en) * 1977-05-31 1981-12-02 Kolehmainen S Selective measurement of somatic and microbial cells
US4906565A (en) * 1986-03-21 1990-03-06 Minnesota Mining And Manufacturing Company Method for the selective detection of microbial nucleotides
CH678065A5 (en) * 1988-04-13 1991-07-31 Hamilton Bonaduz Ag Quantitative determn. of adenosine-tri:phosphate - by a bio-luminescent reaction capable of determining ATP in somatic and/or microbial cells
JP2000157295A (ja) * 1998-11-25 2000-06-13 Kikkoman Corp 生細胞数の測定方法
EP1660508A2 (de) * 2003-08-15 2006-05-31 Celsis Inc. Verwendung von neuen verbindungen zur freisetzung von nukleotiden aus lebenden zellen
CN100507524C (zh) * 2004-04-08 2009-07-01 广东省微生物研究所 微生物抗干扰快速检测方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3408440A (en) * 1966-09-26 1968-10-29 Procter & Gamble Method of preparing substantially cell-free somatic antigens
US3745090A (en) * 1970-08-04 1973-07-10 Nasa Method of detecting and counting bacteria in body fluids
US5004684A (en) * 1987-09-22 1991-04-02 Lumac Bv Method for ATP extraction

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Chung et al. (ATP as a measure of anaerobic sludge digester activity, Water Pollution Control Federation, 1988, Vol 60, pp 107-111) *
Gomori et al. (Preparation of buffers for use in enzyme studies, Methods in Enzymology, 1955, Vol 1, pp 138-146) *
Karamohamed et al. (Bioluminometric method for real-time detection of ATPase activity, 2001, Biotechniques, Vol 31, pp 420-425) *
Maturana et al. (Poly(N-vinylpyrrolidone) as metal ion liquid-liquid extractant, 2000, Polymer Bulletin, Vol 45, pp 425-429) *

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US8993260B2 (en) 2012-05-02 2015-03-31 Charles River Laboratories, Inc. Fluorescence-based viability staining method using a membrane permeable flourescent dye and membrane impermeable fluorescence quencher
US8993259B2 (en) 2012-05-02 2015-03-31 Charles River Laboratories, Inc. Method of viability staining with membrane permeable fluorescent dye and membrane impermeable fluorescence quencher
US9709500B2 (en) 2012-05-02 2017-07-18 Charles River Laboratories, Inc. Optical method for detecting viable microorganisms in a cell sample
US10324036B2 (en) 2012-05-02 2019-06-18 Charles River Laboratories, Inc. Porous planar cell capture system
US10976258B2 (en) 2012-05-02 2021-04-13 Charles River Laboratories, Inc. Porous planar cell capture system and method of use
US20130338350A1 (en) * 2012-06-15 2013-12-19 Ut-Battelle, Llc Method for isolating nucleic acids
US9145553B2 (en) * 2012-06-15 2015-09-29 Ut-Battelle, Llc Method for isolating nucleic acids
JP2017533718A (ja) * 2014-11-13 2017-11-16 スリーエム イノベイティブ プロパティズ カンパニー 試料中の細菌ａｔｐを検出するためのａｔｐ−ジホスホヒドロラーゼを含むキット

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US10196671B2 (en) 2019-02-05
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CN101578374B (zh) 2013-08-14
CN101578374A (zh) 2009-11-11
US20130189717A1 (en) 2013-07-25
ATE533855T1 (de) 2011-12-15
EP2057276B1 (de) 2011-11-16

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