WO2003040406A2 - Diagnosis of clinical infection of a wound - Google Patents

Diagnosis of clinical infection of a wound Download PDF

Info

Publication number
WO2003040406A2
WO2003040406A2 PCT/GB2002/005023 GB0205023W WO03040406A2 WO 2003040406 A2 WO2003040406 A2 WO 2003040406A2 GB 0205023 W GB0205023 W GB 0205023W WO 03040406 A2 WO03040406 A2 WO 03040406A2
Authority
WO
WIPO (PCT)
Prior art keywords
wound
marker
concentration
infection
protease
Prior art date
Application number
PCT/GB2002/005023
Other languages
French (fr)
Other versions
WO2003040406A3 (en
Inventor
Breda Mary Cullen
Original Assignee
Johnson & Johnson Medical 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 Johnson & Johnson Medical Limited filed Critical Johnson & Johnson Medical Limited
Priority to AU2002341191A priority Critical patent/AU2002341191A1/en
Priority to EP02774985A priority patent/EP1453970B1/en
Priority to US10/494,507 priority patent/US7794925B2/en
Priority to DE60230320T priority patent/DE60230320D1/en
Publication of WO2003040406A2 publication Critical patent/WO2003040406A2/en
Publication of WO2003040406A3 publication Critical patent/WO2003040406A3/en

Links

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
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • 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
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • 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
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]

Definitions

  • the present invention relates to a method of predicting or diagnosing clinical infection of a wound comprising measuring the concentration of a marker associated with an inflammatory response in wound fluid, for example the concentration of a neutrophil or macrophage protease.
  • the present invention also relates to devices and kits for use in such methods.
  • Wound healing is a complex dynamic process that results in the restoration of anatomic continuity and function; an ideally healed wound is one that has returned to normal anatomic structure, function and appearance.
  • TNF- ⁇ interleukin-1 and tumour necrosis factor alpha
  • Chronically contaminated wounds all contain a tissue bacterial flora. These bacteria may be indigenous to the patient or might be exogenous to the wound. Closure, or eventual healing of the wound is often based on a physician's ability to control the level of this bacterial flora.
  • Current methods used to identify bacterial infection rely mainly on judgement of the odour and appearance of a wound. With experience, it is possible to identify an infection in a wound by certain chemical signs such as redness or pain. Some clinicians take swabs that are then cultured in the laboratory to identify specific organisms, but this technique takes time. The prior art also describes the use of certain proteases as an indicator of healing status.
  • a prognostic aid that would assist in predicting clinical infection of a wound prior to obvious clinical symptoms of infection.
  • Such a prognostic aid would allow early intervention with suitable treatment (e.g. a topical antimicrobial treatment) before wound chronicity sets in.
  • suitable treatment e.g. a topical antimicrobial treatment
  • diagnostic aid that would assist in the early diagnosis of clinical infection, preferably allowing diagnosis prior to obvious clinical symptoms of infection.
  • wound fluid from wounds that are apparently not clinically infected but which go on to become infected within a few days have high levels of neutrophil elastase activity and may also have high levels of other inflammatory proteases, such macrophage proteases and other neutrophil proteases.
  • concentration of fibronectin fragments may also provide a useful indication of the likelihood of subsequent clinical infection; the presence of such fragments in chronic wounds being mainly due to high levels of elastase activity.
  • a method of predicting or diagnosing clinical infection of a wound comprising measuring the concentration of a marker associated with an inflammatory response in wound fluid, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease.
  • a wound dressing or biosensor comprising components of an assay system for measuring the concentration of a marker associated with an inflammatory response, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease, for use in the manufacture of a medicament for predicting the likelihood of clinical infection of the wound or for diagnosing clinical infection of a wound.
  • neutrophil / macrophage proteases examples include elastase, MMP-9, MMP-8, MMP- 1, MMP-12 and cathepsin G. Further examples include: collagen propeptide, a collagen telopeptide, a protease inhibitor, plasmin, lactate dehydrogenase, a cathepsin, a cytokine, a peroxidase enzyme, a cortisol free radical or a growth factors. More preferably, the marker is a protease enzyme selected from the group consisting of matrix metalloproteinases (e.g. MMP-9), neutrophil elastase, plasmin, low molecular weight gelatinases and latent or active elastases, interleukin converting enzymes or tumor necrosis factor (TNF ⁇ ) converting enzymes .
  • matrix metalloproteinases e.g. MMP-9
  • neutrophil elastase neutrophil elastase
  • plasmin
  • wound fluid refers to any wound exudate or other fluid (preferably substantially not including blood) that is present at the surface of the wound, or that is removed from the wound surface by aspiration, absorption or washing.
  • wound fluid does not refer to blood or tissue plasma remote from the wound site.
  • concentration of more than one marker may be measured. In certain embodiments, the concentrations of at least two, three or four markers are monitored.
  • Measuring the concentration of a marker associated with an inflammatory response in wound fluid allows the likelihood of (or the presence of) clinical infection to be assessed.
  • the step of measuring is preferably carried out on wound fluid that has been removed from the body of the patient, but can also be performed on wound fluid in situ.
  • Any type of wound may be diagnosed for infection according to the method / use of the present invention.
  • the wound may be an acute wound such as an acute traumatic laceration, perhaps resulting from an intentional operative incision, or the wound may be a chronic wound.
  • the method / use of the invention is envisaged as being most useful in predicting or diagnosing clinical infection of a chronic wound.
  • the chronic wound is selected from the group consisting of venous ulcers, pressure sores, decubitis ulcers, diabetic ulcers and chronic ulcers of unknown aetiology.
  • Chronic wound fluids inherently have levels of markers such as neutrophil elastase that are many times the level found in normal, acute wound fluids. Nevertheless, it has now been found that the levels of such markers are further elevated by a substantial amount when the chronic wound will become infected.
  • the prognostic / diagnostic assay is designed so as to provide a correlation between a given concentration of a marker of an inflammatory response and the likelihood of (or presence of) clinical infection.
  • concentration levels of markers of the inflammatory response which are indicative of subsequent progression to clinical infection and/or of the presence of clinical infection.
  • a concentration at least 1.5-, 2-, 2.5-, 3.0-, 3.5-, 4.0-, 4.5-, 5.0-, 5.5-, 6.0-, 7.0-, 8.0- or 9.0-fold the basal level of the marker is considered as begin indicative of subsequent progression to clinical infection.
  • measuring the concentration of a marker associated with an inflammatory response we also include techniques that produce a positive or negative signal if the marker is present at one or more of these concentrations.
  • a concentration at least 1.5-, 2-, 2.5-, 3.0-, 3.5-, 4.0-, 4.5-, 5.0-, 5.5-, 6.0-, 7.0-, 8.0-, 9.0-, 10.0-, 11.0-, 12.0- 14.0-, 16.0-fold the basal level of the marker is considered as begin indicative of the presence of clinical infection.
  • concentration of a marker associated with an inflammatory response we also include techniques that produce a positive or negative signal if the marker is present at one or more of these concentrations.
  • basal level of the marker we include the level of the marker normally associated with a wound which is not clinically infected and which does not subsequently become clinically infected. It will be appreciated that the basal level of the marker may be much higher for a chronic wound than for a normal, acute wound.
  • wound fluid is meant to refer to the exudate that is secreted or discharged by cells in the environment of the wound. This fluid contains cells, both living and dead, and a variety of inflammatory cytokines.
  • concentration of a marker of an inflammatory response is meant the free concentration of the marker in the wound fluid.
  • concentration of the marker may be assessed in situ, or alternatively a sample of wound fluid may be taken as a clinical swab or as a fluid sample.
  • the concentration of the marker of the inflammatory response may be measured by any method known to those of skill in the art. Suitable methods include those utilising chemical or enzyme-linked reactions, or immunological (e.g. ELISA, western blots), spectrophotometric, colorimetric, fluorimetric, or radioactive detection based techniques. In one embodiment the concentration of the marker is measured by a dip-stick type test. Such a test could be used in the community and by the patient allowing easier and earlier diagnosis.
  • a sample of wound fluid must be added to the assay system. Measurement may either be made in situ, or fluid may be removed from the wound for subsequent analysis. The decision as to which method is used will depend upon the type of wound in question.
  • a chnical swab, dressing, "dipstick” or other biosensor device may be applied directly to the surface of the wound.
  • the device should contain the components of the assay system for measuring the concentration of the marker so that the assay reaction may itself proceed in situ. The device can then be removed from the wound and the signal measured by the appropriate means.
  • a physician may not actually require an accurate assessment of the precise concentration of the marker, but may just wish to know whether there is a sufficient concentration of the marker to warrant prophylactic or curative action as necessary. In these cases, visible assessment of the dressing may be sufficient to allow identification of the specific areas of infection. Unnecessary treatment of healthy granulating tissue can then be avoided.
  • a dressing that allows mapping of the infected areas of a wound will be preferable in certain instances.
  • Diagnostic wound mapping sheets that could be adapted to the methods of the present invention are described in GB2323166 (application no. GB 9705081.9), filed on 12th March 1997, the entire content of which is hereby incorporated by reference.
  • Immobilisation of reaction components onto a dipstick, wound mapping sheet or other solid or gel substrate offers the opportunity of performing a more quantitative measurement.
  • the device may be transferred to a spectrometer. Suitable methods of analysis will be apparent to those of skill in the art.
  • Immobilisation of the reaction components to a small biosensor device will also have the advantage that less of the components (such as enzyme and substrate) are needed.
  • the device will thus be less expensive to manufacture than a dressing that needs to have a large surface area in order to allow the mapping of a large wound area.
  • the concentration of the marker may alternatively be measured in an aqueous assay system.
  • Wound fluid may be extracted directly from the environment of the wound or can be washed off the wound using a saline buffer. The resulting solution can then be assayed for the concentration of the marker in, for example, a test tube or in a microassay plate.
  • a test tube or in a microassay plate.
  • Such a method will be preferable for use in cases in which the wound is too small or too inaccessible to allow access of a diagnostic device such as a dipstick.
  • This method has the additional advantage that the wound exudate sample may be diluted.
  • an aqueous assay system is more applicable to use in a laboratory environment, whereas a wound dressing containing the necessary reaction components will be more suitable for use in a hospital or domestic environment.
  • Figure 1 shows the levels of various proteases in diabetic wound fluid. Protease levels including elastase has been corrected for total protein present in the wound fluid sample, thus data is presented as RFU/min/mg protein (relative fluorescence units per minute - rate of change of fluorescence). Patient 8 had 10-fold more elastase activity and within two days was hospitalised for severe cellulitis/infection.
  • the protein binding solution comprises 1 ml Coomassie Brillant Blue stock solution 200mg-Coomassie Brillant Blue G250, Sigma Chemical Co., dissolved in 50 ml ethanol-90%); 2ml orthophosphoric acid (85% w/v); in a final volume of 20 ml with distilled water. This solution was filtered (Whatman #1 filter paper) and used immediately.
  • the protein level in a sample wound fluid was measured by mixing 10- ⁇ l sample or standard with 190- ⁇ l of the protein binding solution in a microtitre well and incubating for 30mins at ambient temperature prior to reading absorbance at 595nm.
  • the concentration of protein was estimated from a standard calibration of BSA (bovine serum albumin prepared in distilled water; Sigma Chemical Co.) ranging from 1.0 to 001 mg/ml.
  • the levels of neutrophil-derived elastase, plasmin and matrix metalloproteinases present in the wound fluid samples were measured spectrofluorimetrically using substrate activity assays.
  • the substrates comprise short peptides synthesised to mimic the appropriate enzyme cleavage site and contain a fluorescent reporter group which is released upon hydrolysis.
  • Enzyme activity was determined by measuring the rate of production of the fluorimetric compound, 7-amino 4-methyl coumarin. Activity was expressed either as relative fluorescence units per minute (RFU/min) or change in fluorescence when corrected for total protein (RFU/min/mg protein). Each sample was tested times 6 and the average value calculated.
  • the substrate was prepared at lOmM-stock concentration, and diluted to a working concentration of 0.5mM in the appropriate assay buffer.
  • the reaction mixture combined in a microtitre well (black, flat bottomed) comprised 5 ⁇ l wound fluid, 175 ⁇ l assay buffer and 20 ⁇ l substrate (final concentration 50 ⁇ M).
  • the microtitre plate was read immediately at 455nm (excitation 383nm) and at timed intervals over the next hour, between readings the plate was covered and incubated at 37°C.
  • Neutrophil-derived elastase-like activity was estimated using the fluorimetric substrate Methoxy-Alanine-Proline-Valine-7-amino 4-methyl coumarin (Bachem UK, Ltd.) solubilised in methanol.
  • the assay buffer required for optimal activity of this enzyme was 0.1M Hepes, pH 7.5 containing 0.5M NaCl and 10% dimethyl sulphoxide.
  • Plasmin-like activity was measured using Methoxysuccinyl-Alanine-Lysine- Phenylalanine-Lysine 7-amino 4-methyl coumarin (Bachem UK, Ltd.) solubilised in lmM- HC1.
  • the assay buffer required for plasmin activity was 25 mM Tris-HCl, pH 8.1 containing 0.5 ⁇ % triton X-100.
  • Matrix metalloproteinase-like activity was estimated utilising the substrate Succinyl- Glycine-Proline-Leucine-Glycine-Proline 7-amino 4-methyl coumarin (Bachem, UK, Ltd.) solubilised in methanol.
  • the assay buffer necessary for maximal MMP activity was 40mM Tris/HCl, pH 7.4 containing 200mM NaCl and lOmM CaCl 2 .

Abstract

A method of predicting or diagnosing clinical infection of a wound comprising measuring the concentration of a maker associated with an inflammatory response in wound fluid, wherein the maker is a fibronectin fragment, a neutrophil protease or a macrophage protease. Also provided is a use of a wound dressing or biosensor comprising components of an assay system for measuring the concentration of a marker associated with an inflammatory response, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease, for use in the manufacture of a medicament for predicting the likelihood of clinical infection of the wound or for diagnosing clinical infection of a wound.

Description

WOUND MONITORING
The present invention relates to a method of predicting or diagnosing clinical infection of a wound comprising measuring the concentration of a marker associated with an inflammatory response in wound fluid, for example the concentration of a neutrophil or macrophage protease. The present invention also relates to devices and kits for use in such methods.
In mammals, injury triggers an organised complex cascade of cellular and biochemical events that result in a healed wound. Wound healing is a complex dynamic process that results in the restoration of anatomic continuity and function; an ideally healed wound is one that has returned to normal anatomic structure, function and appearance.
Infection of wounds by bacteria delays the healing process, since bacteria compete for nutrients and oxygen with macrophages and fibroblasts, whose activity are essential for the healing of the wound. Infection results when bacteria achieve dominance over the systemic and local factors of host resistance. Infection is therefore a manifestation of a disturbed host/bacteria equilibrium in favour of the invading bacteria. This elicits a systemic septic response, and also inhibits the multiple processes involved in wound healing. Lastly, infection can result in a prolonged inflammatory phase and thus slow healing, or may cause further necrosis of the wound. The granulation phase of the healing process will begin only after the infection has subsided.
The persistent presence of bacteria in injured tissue results in the prolonged elevation of proinflammatory cytokines such as interleukin-1 and tumour necrosis factor alpha (TNF- α). This in turn causes increases in the levels of matrix metalloproteinases, a decreased level of tissue inhibitors to the metalloproteinases (TIMP), and a decreased production of growth factors.
Chronically contaminated wounds all contain a tissue bacterial flora. These bacteria may be indigenous to the patient or might be exogenous to the wound. Closure, or eventual healing of the wound is often based on a physician's ability to control the level of this bacterial flora. Current methods used to identify bacterial infection rely mainly on judgement of the odour and appearance of a wound. With experience, it is possible to identify an infection in a wound by certain chemical signs such as redness or pain. Some clinicians take swabs that are then cultured in the laboratory to identify specific organisms, but this technique takes time. The prior art also describes the use of certain proteases as an indicator of healing status.
If clinicians could respond to wound infection as early as possible the infection could be treated topically as opposed to having to use antibiotics. This would also lead to less clinical intervention/hospitalisation and would reduce the use of antibiotics and other complications of infection.
There is thus a long felt need for a prognostic aid that would assist in predicting clinical infection of a wound prior to obvious clinical symptoms of infection. Such a prognostic aid would allow early intervention with suitable treatment (e.g. a topical antimicrobial treatment) before wound chronicity sets in. There is also a need for a diagnostic aid that would assist in the early diagnosis of clinical infection, preferably allowing diagnosis prior to obvious clinical symptoms of infection.
It has been discovered that wound fluid from wounds that are apparently not clinically infected but which go on to become infected within a few days have high levels of neutrophil elastase activity and may also have high levels of other inflammatory proteases, such macrophage proteases and other neutrophil proteases. Similarly, the concentration of fibronectin fragments may also provide a useful indication of the likelihood of subsequent clinical infection; the presence of such fragments in chronic wounds being mainly due to high levels of elastase activity.
According to the present invention, there is provided a method of predicting or diagnosing clinical infection of a wound comprising measuring the concentration of a marker associated with an inflammatory response in wound fluid, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease. Also provided is a use of a wound dressing or biosensor comprising components of an assay system for measuring the concentration of a marker associated with an inflammatory response, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease, for use in the manufacture of a medicament for predicting the likelihood of clinical infection of the wound or for diagnosing clinical infection of a wound.
Examples of neutrophil / macrophage proteases include elastase, MMP-9, MMP-8, MMP- 1, MMP-12 and cathepsin G. Further examples include: collagen propeptide, a collagen telopeptide, a protease inhibitor, plasmin, lactate dehydrogenase, a cathepsin, a cytokine, a peroxidase enzyme, a cortisol free radical or a growth factors. More preferably, the marker is a protease enzyme selected from the group consisting of matrix metalloproteinases ( e.g. MMP-9), neutrophil elastase, plasmin, low molecular weight gelatinases and latent or active elastases, interleukin converting enzymes or tumor necrosis factor (TNFα) converting enzymes .
By "measuring the concentration of a marker associated with an inflammatory response in wound fluid" we include measuring the activity of a marker associated with an inflammatory response in wound fluid. The term "wound fluid" refers to any wound exudate or other fluid (preferably substantially not including blood) that is present at the surface of the wound, or that is removed from the wound surface by aspiration, absorption or washing. The term "wound fluid" does not refer to blood or tissue plasma remote from the wound site.
It will be appreciated that the concentration of more than one marker may be measured. In certain embodiments, the concentrations of at least two, three or four markers are monitored.
Measuring the concentration of a marker associated with an inflammatory response in wound fluid allows the likelihood of (or the presence of) clinical infection to be assessed. The step of measuring is preferably carried out on wound fluid that has been removed from the body of the patient, but can also be performed on wound fluid in situ. Any type of wound may be diagnosed for infection according to the method / use of the present invention. For example, the wound may be an acute wound such as an acute traumatic laceration, perhaps resulting from an intentional operative incision, or the wound may be a chronic wound. The method / use of the invention is envisaged as being most useful in predicting or diagnosing clinical infection of a chronic wound. Preferably, the chronic wound is selected from the group consisting of venous ulcers, pressure sores, decubitis ulcers, diabetic ulcers and chronic ulcers of unknown aetiology. Chronic wound fluids inherently have levels of markers such as neutrophil elastase that are many times the level found in normal, acute wound fluids. Nevertheless, it has now been found that the levels of such markers are further elevated by a substantial amount when the chronic wound will become infected.
According to the present invention, the prognostic / diagnostic assay is designed so as to provide a correlation between a given concentration of a marker of an inflammatory response and the likelihood of (or presence of) clinical infection.
Those skilled in the art will readily be able to determine concentration levels of markers of the inflammatory response, which are indicative of subsequent progression to clinical infection and/or of the presence of clinical infection. Preferably, a concentration at least 1.5-, 2-, 2.5-, 3.0-, 3.5-, 4.0-, 4.5-, 5.0-, 5.5-, 6.0-, 7.0-, 8.0- or 9.0-fold the basal level of the marker is considered as begin indicative of subsequent progression to clinical infection. By "measuring the concentration of a marker associated with an inflammatory response" we also include techniques that produce a positive or negative signal if the marker is present at one or more of these concentrations.
Preferably, a concentration at least 1.5-, 2-, 2.5-, 3.0-, 3.5-, 4.0-, 4.5-, 5.0-, 5.5-, 6.0-, 7.0-, 8.0-, 9.0-, 10.0-, 11.0-, 12.0- 14.0-, 16.0-fold the basal level of the marker is considered as begin indicative of the presence of clinical infection. By "measuring the concentration of a marker associated with an inflammatory response" we also include techniques that produce a positive or negative signal if the marker is present at one or more of these concentrations.
By the "basal level of the marker" we include the level of the marker normally associated with a wound which is not clinically infected and which does not subsequently become clinically infected. It will be appreciated that the basal level of the marker may be much higher for a chronic wound than for a normal, acute wound.
With regard to the concentration of neutrophil elastase which is associated with subsequent progression to clinical infection, our data indicates that an enzyme activity of at least 3,000mg/min is indicative of the wound subsequently becoming clinically infected.
As used herein, the term wound fluid is meant to refer to the exudate that is secreted or discharged by cells in the environment of the wound. This fluid contains cells, both living and dead, and a variety of inflammatory cytokines.
By the "concentration of a marker of an inflammatory response" is meant the free concentration of the marker in the wound fluid. The concentration of the marker may be assessed in situ, or alternatively a sample of wound fluid may be taken as a clinical swab or as a fluid sample.
The concentration of the marker of the inflammatory response may be measured by any method known to those of skill in the art. Suitable methods include those utilising chemical or enzyme-linked reactions, or immunological (e.g. ELISA, western blots), spectrophotometric, colorimetric, fluorimetric, or radioactive detection based techniques. In one embodiment the concentration of the marker is measured by a dip-stick type test. Such a test could be used in the community and by the patient allowing easier and earlier diagnosis.
To allow measurement of concentration of a marker of the inflammatory response in a wound, a sample of wound fluid must be added to the assay system. Measurement may either be made in situ, or fluid may be removed from the wound for subsequent analysis. The decision as to which method is used will depend upon the type of wound in question.
For example, in the case of surface-exposed wounds, a chnical swab, dressing, "dipstick" or other biosensor device may be applied directly to the surface of the wound. The device should contain the components of the assay system for measuring the concentration of the marker so that the assay reaction may itself proceed in situ. The device can then be removed from the wound and the signal measured by the appropriate means. In many cases, a physician may not actually require an accurate assessment of the precise concentration of the marker, but may just wish to know whether there is a sufficient concentration of the marker to warrant prophylactic or curative action as necessary. In these cases, visible assessment of the dressing may be sufficient to allow identification of the specific areas of infection. Unnecessary treatment of healthy granulating tissue can then be avoided.
A dressing that allows mapping of the infected areas of a wound will be preferable in certain instances. Diagnostic wound mapping sheets that could be adapted to the methods of the present invention are described in GB2323166 (application no. GB 9705081.9), filed on 12th March 1997, the entire content of which is hereby incorporated by reference.
Immobilisation of reaction components onto a dipstick, wound mapping sheet or other solid or gel substrate offers the opportunity of performing a more quantitative measurement. For example, in the case of a reaction linked to the generation of a colour the device may be transferred to a spectrometer. Suitable methods of analysis will be apparent to those of skill in the art.
Immobilisation of the reaction components to a small biosensor device will also have the advantage that less of the components (such as enzyme and substrate) are needed. The device will thus be less expensive to manufacture than a dressing that needs to have a large surface area in order to allow the mapping of a large wound area.
Methods for the incorporation of the components of the assay reaction onto a clinical dressing, "dipstick", sheet or other biosensor are routine in the art. See for example Fagerstam and Karlsson (1994) Immunochemistry, 949-970.
The concentration of the marker may alternatively be measured in an aqueous assay system. Wound fluid may be extracted directly from the environment of the wound or can be washed off the wound using a saline buffer. The resulting solution can then be assayed for the concentration of the marker in, for example, a test tube or in a microassay plate. Such a method will be preferable for use in cases in which the wound is too small or too inaccessible to allow access of a diagnostic device such as a dipstick. This method has the additional advantage that the wound exudate sample may be diluted.
It will be clear that an aqueous assay system is more applicable to use in a laboratory environment, whereas a wound dressing containing the necessary reaction components will be more suitable for use in a hospital or domestic environment.
Specific embodiments of the present invention will now be described in more detail, by way of example, with reference to the accompanying drawings, in which: -
BRIEF DESCRIPTION OF THE FIGURE
Figure 1 shows the levels of various proteases in diabetic wound fluid. Protease levels including elastase has been corrected for total protein present in the wound fluid sample, thus data is presented as RFU/min/mg protein (relative fluorescence units per minute - rate of change of fluorescence). Patient 8 had 10-fold more elastase activity and within two days was hospitalised for severe cellulitis/infection.
EXAMPLES
Clinical study and patient selection
All patients enrolled in this study had diabetic foot ulcers of at least 30 days duration and a surface area of at least 1 cm2. Patients were excluded if the target wound showed any signs of infection or if exposed bone with positive osteomyelitis was observed. Additional exclusion criteria included concomitant conditions or treatments that may have interfered with wound healing and a history of non-compliance that would make it unlikely that a patient would complete the study. Nine patients meeting these study criteria were enrolled, and wound fluid collected. Informed consent was obtained from all patients or their authorised representatives prior to study enrolment and the protocol was approved by the Ethics Committee at the participating study centre prior to the commencement of the study. The study was conducted in accordance with both the Declaration of Helsinki and Good Clinical Practice.
Protein Assay
Total protein present in each extracted wound fluid sample was determined using the Bradford protein assay. The protein binding solution comprises 1 ml Coomassie Brillant Blue stock solution 200mg-Coomassie Brillant Blue G250, Sigma Chemical Co., dissolved in 50 ml ethanol-90%); 2ml orthophosphoric acid (85% w/v); in a final volume of 20 ml with distilled water. This solution was filtered (Whatman #1 filter paper) and used immediately. The protein level in a sample wound fluid was measured by mixing 10-μl sample or standard with 190-μl of the protein binding solution in a microtitre well and incubating for 30mins at ambient temperature prior to reading absorbance at 595nm. The concentration of protein was estimated from a standard calibration of BSA (bovine serum albumin prepared in distilled water; Sigma Chemical Co.) ranging from 1.0 to 001 mg/ml.
Protease activity assays
The levels of neutrophil-derived elastase, plasmin and matrix metalloproteinases present in the wound fluid samples were measured spectrofluorimetrically using substrate activity assays. The substrates comprise short peptides synthesised to mimic the appropriate enzyme cleavage site and contain a fluorescent reporter group which is released upon hydrolysis. Enzyme activity was determined by measuring the rate of production of the fluorimetric compound, 7-amino 4-methyl coumarin. Activity was expressed either as relative fluorescence units per minute (RFU/min) or change in fluorescence when corrected for total protein (RFU/min/mg protein). Each sample was tested times 6 and the average value calculated. The substrate was prepared at lOmM-stock concentration, and diluted to a working concentration of 0.5mM in the appropriate assay buffer. The reaction mixture, combined in a microtitre well (black, flat bottomed) comprised 5μl wound fluid, 175μl assay buffer and 20μl substrate (final concentration 50μM). The microtitre plate was read immediately at 455nm (excitation 383nm) and at timed intervals over the next hour, between readings the plate was covered and incubated at 37°C. Neutrophil-derived elastase-like activity was estimated using the fluorimetric substrate Methoxy-Alanine-Proline-Valine-7-amino 4-methyl coumarin (Bachem UK, Ltd.) solubilised in methanol. The assay buffer required for optimal activity of this enzyme was 0.1M Hepes, pH 7.5 containing 0.5M NaCl and 10% dimethyl sulphoxide.
Plasmin-like activity was measured using Methoxysuccinyl-Alanine-Lysine- Phenylalanine-Lysine 7-amino 4-methyl coumarin (Bachem UK, Ltd.) solubilised in lmM- HC1. The assay buffer required for plasmin activity was 25 mM Tris-HCl, pH 8.1 containing 0.5~% triton X-100.
Matrix metalloproteinase-like activity was estimated utilising the substrate Succinyl- Glycine-Proline-Leucine-Glycine-Proline 7-amino 4-methyl coumarin (Bachem, UK, Ltd.) solubilised in methanol. The assay buffer necessary for maximal MMP activity was 40mM Tris/HCl, pH 7.4 containing 200mM NaCl and lOmM CaCl2.
The results of the assays are shown in Figure 1. Patient 8 had 10-fold more elastase activity and within two days was hospitalised for severe cellulitis/infection.
The above example has been described for the purpose of illustration only. Many other embodiments falling within the scope of the accompanying claims will be apparent to the skilled reader.

Claims

1. A method of predicting or diagnosing clinical infection of a wound comprising measuring the concentration of a marker associated with an inflammatory response in wound fluid, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease.
2. Use of a wound dressing or biosensor comprising components of an assay system for measuring the concentration of a marker associated with an inflammatory response, wherein the marker is a fibronectin fragment, a neutrophil protease or a macrophage protease, for use in the manufacture of a medicament for predicting the likelihood of clinical infection of the wound or for diagnosing clinical infection of a wound.
3. The method of claim 1 or the use of claim 2 wherein the marker is neutrophil elastase.
4. The method or use according to any one of claims 1 to 3 wherein the wound is a chronic wound.
5. The method or use according to claim 4 wherein the chronic wound is a chronic ulcer such as a dermal ulcer, venous ulcer, pressure sore or decubitis ulcer.
6. The method or use according to any one of claims 1 to 5 wherein the concentration of the marker is measured using an immunological, spectrophotometric, colorimetric, fluorimetric, or radioactive detection based technique.
7. The method or use according to any one of claims 1 to 6 wherein the concentration of the marker is measured by a dip-stick type test.
PCT/GB2002/005023 2001-11-05 2002-11-05 Diagnosis of clinical infection of a wound WO2003040406A2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU2002341191A AU2002341191A1 (en) 2001-11-05 2002-11-05 Diagnosis of clinical infection of a wound
EP02774985A EP1453970B1 (en) 2001-11-05 2002-11-05 Diagnosis of clinical infection of a wound
US10/494,507 US7794925B2 (en) 2001-11-05 2002-11-05 Diagnosis of clinical infection of a wound
DE60230320T DE60230320D1 (en) 2001-11-05 2002-11-05 DIAGNOSIS OF A CLINICAL INFECTION OF A WOUNCE

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0126534A GB2381452B (en) 2001-11-05 2001-11-05 Wound monitoring
GB0126534.7 2001-11-05

Publications (2)

Publication Number Publication Date
WO2003040406A2 true WO2003040406A2 (en) 2003-05-15
WO2003040406A3 WO2003040406A3 (en) 2003-10-16

Family

ID=9925173

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2002/005023 WO2003040406A2 (en) 2001-11-05 2002-11-05 Diagnosis of clinical infection of a wound

Country Status (8)

Country Link
US (1) US7794925B2 (en)
EP (1) EP1453970B1 (en)
AT (1) ATE417125T1 (en)
AU (1) AU2002341191A1 (en)
DE (1) DE60230320D1 (en)
ES (1) ES2316621T3 (en)
GB (1) GB2381452B (en)
WO (1) WO2003040406A2 (en)

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005121357A2 (en) * 2004-06-11 2005-12-22 Ethicon, Inc. Diagnosis and prognosis of wound infection by measurement of a phospholipase a2 in wound fluid
EP2118653A2 (en) * 2006-12-07 2009-11-18 University Of Florida Research Foundation, Inc. Materials and methods for efficient and accurate detection of analytes
US8012698B2 (en) 2007-03-30 2011-09-06 Systagenix Wound Management (Us), Inc. Diagnostic markers of wound infection
EP2446049A2 (en) * 2009-06-26 2012-05-02 University of Florida Research Foundation, Incorporated Rapid bed-side measurement of neutrophil elastase activity in biological fluids
WO2012104620A2 (en) * 2011-01-31 2012-08-09 Systagenix Wound Management Ip Co. B.V. Wound prognosis
WO2013041879A3 (en) * 2011-09-23 2013-05-30 Systagenix Wound Management Ip Co. B.V. Wound prognosis
US8609358B2 (en) * 2003-11-03 2013-12-17 Systagenix Wound Management (Us), Inc. Methods, peptides and biosensors useful for detecting a broad spectrum of bacteria
US20140024106A1 (en) * 2004-09-17 2014-01-23 Systagenix Wound Management (Us), Inc. Monitoring of wounds by measurement of protease and protease inhibitor levels in wound fluids
US8685639B2 (en) 2004-06-11 2014-04-01 Woundcheck Laboratories (US), Inc. Diagnosis and prognosis of wound infection by measurement of a phospholipase A2 in wound fluid
US9024751B2 (en) 2012-04-12 2015-05-05 Elwha Llc Dormant to active appurtenances for reporting information regarding wound dressings
US9084530B2 (en) 2012-04-12 2015-07-21 Elwha Llc Computational methods and systems for reporting information regarding appurtenances to wound dressings
WO2017182765A1 (en) * 2016-04-18 2017-10-26 Swansea University Method of detecting bacterial infection in a biological sample
US10130518B2 (en) 2012-04-12 2018-11-20 Elwha Llc Appurtenances including sensors for reporting information regarding wound dressings
US10158928B2 (en) 2012-04-12 2018-12-18 Elwha Llc Appurtenances for reporting information regarding wound dressings
US10226212B2 (en) 2012-04-12 2019-03-12 Elwha Llc Appurtenances to cavity wound dressings
US10265219B2 (en) 2012-04-12 2019-04-23 Elwha Llc Wound dressing monitoring systems including appurtenances for wound dressings
EP3901277A1 (en) 2020-04-24 2021-10-27 Paul Hartmann AG Method and device for assessing a status of a wound of a patient
EP3901276A1 (en) 2020-04-24 2021-10-27 Paul Hartmann AG Method and device for assessing a status of a wound of a patient

Families Citing this family (61)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6036924A (en) 1997-12-04 2000-03-14 Hewlett-Packard Company Cassette of lancet cartridges for sampling blood
US6391005B1 (en) 1998-03-30 2002-05-21 Agilent Technologies, Inc. Apparatus and method for penetration with shaft having a sensor for sensing penetration depth
US8641644B2 (en) 2000-11-21 2014-02-04 Sanofi-Aventis Deutschland Gmbh Blood testing apparatus having a rotatable cartridge with multiple lancing elements and testing means
US9795747B2 (en) 2010-06-02 2017-10-24 Sanofi-Aventis Deutschland Gmbh Methods and apparatus for lancet actuation
US9427532B2 (en) 2001-06-12 2016-08-30 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US7025774B2 (en) 2001-06-12 2006-04-11 Pelikan Technologies, Inc. Tissue penetration device
US8337419B2 (en) 2002-04-19 2012-12-25 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US7981056B2 (en) 2002-04-19 2011-07-19 Pelikan Technologies, Inc. Methods and apparatus for lancet actuation
US9226699B2 (en) 2002-04-19 2016-01-05 Sanofi-Aventis Deutschland Gmbh Body fluid sampling module with a continuous compression tissue interface surface
US7682318B2 (en) 2001-06-12 2010-03-23 Pelikan Technologies, Inc. Blood sampling apparatus and method
EP1404233B1 (en) 2001-06-12 2009-12-02 Pelikan Technologies Inc. Self optimizing lancing device with adaptation means to temporal variations in cutaneous properties
ATE497731T1 (en) 2001-06-12 2011-02-15 Pelikan Technologies Inc DEVICE FOR INCREASING THE SUCCESS RATE OF BLOOD YIELD OBTAINED BY A FINGER PICK
EP1404235A4 (en) 2001-06-12 2008-08-20 Pelikan Technologies Inc Method and apparatus for lancet launching device integrated onto a blood-sampling cartridge
JP4149911B2 (en) 2001-06-12 2008-09-17 ペリカン テクノロジーズ インコーポレイテッド Electric lancet actuator
US7291117B2 (en) 2002-04-19 2007-11-06 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7674232B2 (en) 2002-04-19 2010-03-09 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US9795334B2 (en) 2002-04-19 2017-10-24 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
US8267870B2 (en) 2002-04-19 2012-09-18 Sanofi-Aventis Deutschland Gmbh Method and apparatus for body fluid sampling with hybrid actuation
US7229458B2 (en) 2002-04-19 2007-06-12 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7491178B2 (en) 2002-04-19 2009-02-17 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US8784335B2 (en) 2002-04-19 2014-07-22 Sanofi-Aventis Deutschland Gmbh Body fluid sampling device with a capacitive sensor
US7901362B2 (en) 2002-04-19 2011-03-08 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7297122B2 (en) 2002-04-19 2007-11-20 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7648468B2 (en) 2002-04-19 2010-01-19 Pelikon Technologies, Inc. Method and apparatus for penetrating tissue
US7547287B2 (en) 2002-04-19 2009-06-16 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US9248267B2 (en) 2002-04-19 2016-02-02 Sanofi-Aventis Deustchland Gmbh Tissue penetration device
US7232451B2 (en) 2002-04-19 2007-06-19 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US9314194B2 (en) 2002-04-19 2016-04-19 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US7226461B2 (en) 2002-04-19 2007-06-05 Pelikan Technologies, Inc. Method and apparatus for a multi-use body fluid sampling device with sterility barrier release
US7371247B2 (en) 2002-04-19 2008-05-13 Pelikan Technologies, Inc Method and apparatus for penetrating tissue
US7976476B2 (en) 2002-04-19 2011-07-12 Pelikan Technologies, Inc. Device and method for variable speed lancet
US7331931B2 (en) 2002-04-19 2008-02-19 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7717863B2 (en) 2002-04-19 2010-05-18 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US8221334B2 (en) 2002-04-19 2012-07-17 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
US7909778B2 (en) 2002-04-19 2011-03-22 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US8579831B2 (en) 2002-04-19 2013-11-12 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
US7892183B2 (en) 2002-04-19 2011-02-22 Pelikan Technologies, Inc. Method and apparatus for body fluid sampling and analyte sensing
US8702624B2 (en) 2006-09-29 2014-04-22 Sanofi-Aventis Deutschland Gmbh Analyte measurement device with a single shot actuator
US7175642B2 (en) 2002-04-19 2007-02-13 Pelikan Technologies, Inc. Methods and apparatus for lancet actuation
US8574895B2 (en) 2002-12-30 2013-11-05 Sanofi-Aventis Deutschland Gmbh Method and apparatus using optical techniques to measure analyte levels
US7850621B2 (en) 2003-06-06 2010-12-14 Pelikan Technologies, Inc. Method and apparatus for body fluid sampling and analyte sensing
WO2006001797A1 (en) 2004-06-14 2006-01-05 Pelikan Technologies, Inc. Low pain penetrating
EP1671096A4 (en) 2003-09-29 2009-09-16 Pelikan Technologies Inc Method and apparatus for an improved sample capture device
WO2005037095A1 (en) 2003-10-14 2005-04-28 Pelikan Technologies, Inc. Method and apparatus for a variable user interface
US7822454B1 (en) 2005-01-03 2010-10-26 Pelikan Technologies, Inc. Fluid sampling device with improved analyte detecting member configuration
US8668656B2 (en) 2003-12-31 2014-03-11 Sanofi-Aventis Deutschland Gmbh Method and apparatus for improving fluidic flow and sample capture
EP1751546A2 (en) 2004-05-20 2007-02-14 Albatros Technologies GmbH & Co. KG Printable hydrogel for biosensors
WO2005120365A1 (en) 2004-06-03 2005-12-22 Pelikan Technologies, Inc. Method and apparatus for a fluid sampling device
GB0413001D0 (en) * 2004-06-10 2004-07-14 Johnson & Johnson Medical Ltd Diagnostic and prognostic
US8652831B2 (en) 2004-12-30 2014-02-18 Sanofi-Aventis Deutschland Gmbh Method and apparatus for analyte measurement test time
GB0520405D0 (en) * 2005-10-07 2005-11-16 Imp College Innovations Ltd Biological agents and method
GB0806186D0 (en) * 2008-04-04 2008-05-14 Axis Shield Diagnostics Ltd Method
GB2459098A (en) * 2008-04-08 2009-10-14 Ethicon Inc Genetic markers of wound development
WO2009126900A1 (en) 2008-04-11 2009-10-15 Pelikan Technologies, Inc. Method and apparatus for analyte detecting device
CN104939806B (en) 2008-05-20 2021-12-10 大学健康网络 Apparatus and method for fluorescence-based imaging and monitoring
US9375169B2 (en) 2009-01-30 2016-06-28 Sanofi-Aventis Deutschland Gmbh Cam drive for managing disposable penetrating member actions with a single motor and motor and control system
WO2010133589A1 (en) * 2009-05-18 2010-11-25 Technische Universität Graz Method for detecting a wound infection
US8965476B2 (en) 2010-04-16 2015-02-24 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US10438356B2 (en) 2014-07-24 2019-10-08 University Health Network Collection and analysis of data for diagnostic purposes
JP7183146B2 (en) * 2016-03-30 2022-12-05 コンバテック・テクノロジーズ・インコーポレイテッド Detection of microbial infection in wounds
UY37176A (en) 2016-03-30 2017-10-31 Convatec Technologies Inc MODIFIED DRESSES FOR WOUNDS

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0886140A1 (en) * 1997-06-09 1998-12-23 Ethicon, Inc. Chronic wound model
WO2000008203A1 (en) * 1998-08-05 2000-02-17 Johnson & Johnson Medical Limited Method of monitoring bacterial contamination of a wound

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2323166B (en) 1997-03-12 2001-04-11 Johnson & Johnson Medical Method and apparatus for mapping the condition of a wound
RU2146367C1 (en) * 1997-07-22 2000-03-10 Гуменюк Сергей Евгеньевич Method for predicting wound development process course
US5912114A (en) * 1997-09-12 1999-06-15 Johnson & Johnson Medical, Inc. Wound diagnosis by quantitating cortisol in wound fluids
CA2403371A1 (en) * 2000-02-10 2001-08-16 Timothy F. Murphy A method for detecting bacterial exacerbations of chronic lung disease

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0886140A1 (en) * 1997-06-09 1998-12-23 Ethicon, Inc. Chronic wound model
WO2000008203A1 (en) * 1998-08-05 2000-02-17 Johnson & Johnson Medical Limited Method of monitoring bacterial contamination of a wound

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
ADACHI Y ET AL: "Lipopolysaccharide increases fibronectin production and release from cultured lung fibroblasts partially through proteolytic activity." JOURNAL OF LABORATORY AND CLINICAL MEDICINE, vol. 127, no. 5, 1996, pages 448-455, XP009015383 ISSN: 0022-2143 *
BRATSCHITSCH G ET AL: "Clinical benefit of PMN-elastase as a determinant of inflammation signs for prospective wound healing monitoring after surgical emergency management" ACTA CHIRURGICA AUSTRIACA 1997 AUSTRIA, vol. 29, no. SUPPL. 133, 1997, pages 29-31, XP009015252 ISSN: 0001-544X *
HOFER H P ET AL: "Released PMN elastase: An indicator of postsurgical uneventful wound healing and early inflammatory complications. A contribution to the search for objective criteria in wound healing monitoring." INJURY, vol. 26, no. 2, 1995, pages 103-106, XP002250734 ISSN: 0020-1383 *
NWOMEH B C ET AL: "PHYSIOLOGY OF THE CHRONIC WOUND" CLINICS IN PLASTIC SURGERY, W.B.SAUNDERS CO., LONDON, GB, vol. 25, no. 3, July 1998 (1998-07), pages 341-356, XP008019702 ISSN: 0094-1298 *
TARLTON J F ET AL: "Postsurgical wound progression monitored by temporal changes in the expression of matrix metalloproteinase-9." BRITISH JOURNAL OF DERMATOLOGY, vol. 137, no. 4, 1997, pages 506-516, XP009015255 ISSN: 0007-0963 *
YAGER ET L D R: "Ability of chronic wound fluids to degrade peptide growth factors is associated with increased levels of elastase activity and diminished levels of proteinase inhibitors" WOUND REPAIR AND REGENERATION, MOSBY-YEAR BOOK, ST. LOUIS, MO, US, vol. 1, no. 5, 1997, pages 23-32, XP002080317 ISSN: 1067-1927 *

Cited By (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8609358B2 (en) * 2003-11-03 2013-12-17 Systagenix Wound Management (Us), Inc. Methods, peptides and biosensors useful for detecting a broad spectrum of bacteria
WO2005121357A3 (en) * 2004-06-11 2006-06-01 Ethicon Inc Diagnosis and prognosis of wound infection by measurement of a phospholipase a2 in wound fluid
WO2005121357A2 (en) * 2004-06-11 2005-12-22 Ethicon, Inc. Diagnosis and prognosis of wound infection by measurement of a phospholipase a2 in wound fluid
US8685639B2 (en) 2004-06-11 2014-04-01 Woundcheck Laboratories (US), Inc. Diagnosis and prognosis of wound infection by measurement of a phospholipase A2 in wound fluid
US20140024106A1 (en) * 2004-09-17 2014-01-23 Systagenix Wound Management (Us), Inc. Monitoring of wounds by measurement of protease and protease inhibitor levels in wound fluids
EP2118653A2 (en) * 2006-12-07 2009-11-18 University Of Florida Research Foundation, Inc. Materials and methods for efficient and accurate detection of analytes
EP2118653A4 (en) * 2006-12-07 2009-12-09 Univ Florida Materials and methods for efficient and accurate detection of analytes
US8012698B2 (en) 2007-03-30 2011-09-06 Systagenix Wound Management (Us), Inc. Diagnostic markers of wound infection
EP2446049A4 (en) * 2009-06-26 2013-03-06 Univ Florida Rapid bed-side measurement of neutrophil elastase activity in biological fluids
EP2446049A2 (en) * 2009-06-26 2012-05-02 University of Florida Research Foundation, Incorporated Rapid bed-side measurement of neutrophil elastase activity in biological fluids
US10577639B2 (en) 2009-06-26 2020-03-03 University Of Florida Research Foundation, Inc. Rapid bed-side measurement of neutrophil elastase activity in biological fluids
US9932622B2 (en) 2011-01-31 2018-04-03 Woundchek Laboratories B.V. Wound prognosis
WO2012104620A3 (en) * 2011-01-31 2012-11-01 Systagenix Wound Management Ip Co. B.V. Wound prognosis
US20140045761A1 (en) * 2011-01-31 2014-02-13 Systagenix Wound Management IP CO. BV Wound prognosis
WO2012104620A2 (en) * 2011-01-31 2012-08-09 Systagenix Wound Management Ip Co. B.V. Wound prognosis
EP2896963A1 (en) * 2011-01-31 2015-07-22 Woundchek Laboratories B.V. Wound prognosis
WO2013041879A3 (en) * 2011-09-23 2013-05-30 Systagenix Wound Management Ip Co. B.V. Wound prognosis
US9024751B2 (en) 2012-04-12 2015-05-05 Elwha Llc Dormant to active appurtenances for reporting information regarding wound dressings
US10265219B2 (en) 2012-04-12 2019-04-23 Elwha Llc Wound dressing monitoring systems including appurtenances for wound dressings
US9084530B2 (en) 2012-04-12 2015-07-21 Elwha Llc Computational methods and systems for reporting information regarding appurtenances to wound dressings
US9451340B2 (en) 2012-04-12 2016-09-20 Elwha Llc Computational methods and systems for reporting information regarding appurtenances to wound dressings
US10130518B2 (en) 2012-04-12 2018-11-20 Elwha Llc Appurtenances including sensors for reporting information regarding wound dressings
US10158928B2 (en) 2012-04-12 2018-12-18 Elwha Llc Appurtenances for reporting information regarding wound dressings
US10226212B2 (en) 2012-04-12 2019-03-12 Elwha Llc Appurtenances to cavity wound dressings
US9510781B2 (en) 2012-04-12 2016-12-06 Elwha Llc Dormant to active appurtenances for reporting information regarding wound dressings
WO2017182765A1 (en) * 2016-04-18 2017-10-26 Swansea University Method of detecting bacterial infection in a biological sample
US11453905B2 (en) 2016-04-18 2022-09-27 Trubac, LTD. Method of detecting bacterial infection in a biological sample
EP3901277A1 (en) 2020-04-24 2021-10-27 Paul Hartmann AG Method and device for assessing a status of a wound of a patient
EP3901276A1 (en) 2020-04-24 2021-10-27 Paul Hartmann AG Method and device for assessing a status of a wound of a patient
WO2021214259A1 (en) 2020-04-24 2021-10-28 Paul Hartmann Ag Method and device for assessing a status of a wound of a patient
WO2021214068A1 (en) 2020-04-24 2021-10-28 Paul Hartmann Ag Method and device for assessing a status of a wound of a patient

Also Published As

Publication number Publication date
GB2381452A (en) 2003-05-07
DE60230320D1 (en) 2009-01-22
GB2381452B (en) 2005-08-10
US7794925B2 (en) 2010-09-14
US20050079542A1 (en) 2005-04-14
EP1453970B1 (en) 2008-12-10
ATE417125T1 (en) 2008-12-15
ES2316621T3 (en) 2009-04-16
AU2002341191A1 (en) 2003-05-19
GB0126534D0 (en) 2002-01-02
EP1453970A2 (en) 2004-09-08
WO2003040406A3 (en) 2003-10-16

Similar Documents

Publication Publication Date Title
US7794925B2 (en) Diagnosis of clinical infection of a wound
Ågren et al. Matrix metalloproteinase 9 level predicts optimal collagen deposition during early wound repair in humans
EP2896963B1 (en) Wound prognosis
US20140024106A1 (en) Monitoring of wounds by measurement of protease and protease inhibitor levels in wound fluids
US8497085B2 (en) Method for detecting a wound infection
US8685639B2 (en) Diagnosis and prognosis of wound infection by measurement of a phospholipase A2 in wound fluid
WO2004086043A1 (en) Prediction and detection of wound infection
EP1756298B1 (en) Diagnosis and prognosis of wound infection by measurement of a phospholipase a2 in wound fluid
US20200392557A1 (en) Methods of prediction of wound healing
WO2009122188A2 (en) Wound infection monitoring method
US9404931B2 (en) Measurement of cytoskeletal proteins and uses thereof in diagnosis, prognosis and therapy
GB2430031A (en) Diagnostic markers of wound infection
CN1394282A (en) Diagnostic kit for schizophrenia
GB2399881A (en) Diagnosing wound infections
GB2430029A (en) Diagnostic markers of wound infection

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SI SK SL TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LU MC NL PT SE SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2002774985

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 10494507

Country of ref document: US

WWP Wipo information: published in national office

Ref document number: 2002774985

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP