Diagnostics

Patent metadata

Field	Value
Publication number	US-2017184570-A1
Application number	US-201515324316-A
Country	US
Kind code	A1
Filing date	Jul 8, 2015
Priority date	Jul 8, 2014
Publication date	Jun 29, 2017
Grant date	—

Abstract

The present invention relates to the detection or diagnosis of infection or inflammation in a patient. In particular, the invention provides methods that allow the detection of markers in a sample from a patient or subject, such as a blood sample, which will indicate whether the patient or subject has an infection or has an inflammatory response.

First claim

1 . A method of detecting infection or inflammation in a mammalian subject, the method comprising
(a) detecting the amount of at least one marker that is released from an immune ell on activation in a sample from said subject, (b) contacting the said sample with a stimulator of immune cells, and (c) determining whether the said stimulus leads to a change in the amount of the said marker(s) in the sample.

2 . A method of detecting infection or inflammation in a mammalian subject, the method comprising
(a) detecting the amount of at least one marker selected from glutamate, lactate, ATP, D-serine, acetylcholine and adenosine in a sample from said subject, (b) contacting the said sample with a stimulator of immune cells, and (c) determining whether the said stimulus leads to a change in the amount of the said marker(s) in the sample.

3 . A method of detecting infection or inflammation in a mammalian subject, the method comprising
(a) detecting the amount of at least one marker selected from glutamate, lactate, ATP, D-serine, acetylcholine and adenosine in a sample from said subject and (b) comparing the amount of the marker(s) of (a) with the amount of the same marker(s) in a control sample, wherein an increased or decreased amount of said marker(s) in the sample from the subject, when compared to the control sample, is indicative of the presence of inflammation or infection.

4 . The method of any one of the preceding claims wherein the marker is selected from glutamate, ATP, D-serine, acetylcholine and adenosine.

5 . A method according to claim 1 or 2 comprising comparing the said change in the amount of the marker(s) with the change in the amount of said marker(s) which is seen when said stimulus is applied to a control sample, optionally wherein an increased or decreased response of said marker(s) to the stimulus in the sample from the subject, when compared to the response to the stimulus in the control sample, is indicative of the presence of inflammation or infection.

6 . A method according to claim 5 wherein the change in the amount of the said marker(s) which is seen when the said stimulus is applied to a control sample, optionally a control sample from a normal subject, is used as a control value.

7 . A method according to claim 6 wherein the sample is whole blood and the control value for the said change is a change in concentration of any one of the said marker(s) in the range from around 1 μM to around 6 μM or from around 1 μM to around 10 μM.

8 . A method according to any one of the preceding claims wherein the amount of said marker(s) is tested within 5 minutes of contacting the sample with said stimulus

9 . A method according to any one of the preceding claims wherein the infection is a pathogenic infection, a bacterial infection, a viral infection, a fungal infection and/or sepsis.

10 . A method according to any one of the preceding claims wherein the sample is an in vivo, in vitro or ex vivo sample.

11 . A method according to any one of the previous claims wherein said method is used to monitor the relative amounts of said marker(s) in said subject at two or more different points in time, optionally wherein said control sample is an earlier sample from the subject and a change in the amount of said marker(s) when compared to said control indicates that the subject has developed an infection or inflammation.

12 . A method according to any one of the previous claims wherein said sample is in vivo or ex vivo and said method is used to monitor changes in the amount of said marker(s) in said subject in real-time.

13 . A method according to any one of the preceding claims, further comprising administering a therapeutic agent that treats infection and/or inflammation to a subject that has been identified as having infection or inflammation by said method.

14 . A method according to any one of the preceding claims wherein the sample is a sample of whole blood, urine, CSF or synovial fluid.

15 . A method according to any one of the preceding claims wherein the sample comprises neutrophils.

16 . A method according to any one of the preceding claims wherein said marker(s) are detected using one or more biosensors, wherein each biosensor comprises
(a) a biological element that is capable of recognising said marker(s) and (b) an electrode capable of detecting a reaction caused by the recognition of said marker(s) by the biological element.

17 . A method according to any one of the preceding claims wherein detection of glutamate is carried out using a biosensor comprising glutamate oxidase, optionally wherein said glutamate oxidase catalyses a reaction of glutamate in the sample to form 2-oxoglutarate+NH 3 +H 2 O 2 , and wherein an electrode in said biosensor detects said production of H 2 O 2 .

18 . A method according to any one of the preceding claims wherein detection of lactate is carried out using a biosensor comprising lactate oxidase,
optionally wherein said lactate oxidase catalyses a reaction of lactate in the sample to form pyruvate+H 2 O 2 and wherein an electrode in said biosensor detects said production of H 2 O 2

19 . A method according to any one of the preceding claims wherein detection of ATP is carried out using a biosensor comprising glycerol kinase and glycerol-3-phosphate oxidase,
optionally wherein said glycerol kinase and glycerol-3-phosphate oxidase catalyse a reaction of ATP in the sample to form glycerine phosphate+H 2 O 2 , and wherein an electrode in said biosensor detects said production of H 2 O 2 .

20 . A method according to any one of the preceding claims wherein detection of acetylcholine is carried out using a biosensor comprising acetylcholine esterase and choline oxidase,
optionally wherein said acetylcholine esterase and choline oxidase catalyse a reaction of acetylcholine in the sample to form betaine aldehyde+H 2 O 2 , and wherein an electrode in said biosensor detects said production of H 2 O 2

21 . A method according to any one of the preceding claims wherein detection of D-serine is carried out using a biosensor comprising D-amino acid oxidase, optionally wherein said D-amino acid oxidase catalyses a reaction of D-serine in the sample to form 2-oxo-3-hydroxypropionate+NH 3 +H 2 O 2 , and wherein an electrode in said biosensor detects said production of H 2 O 2 .

22 . A method according to any one of the preceding claims wherein detection of adenosine is carried out using a biosensor comprising adenosine deaminase, nucleoside phosphorylase and xanthine oxidase,
optionally wherein said adenosine deaminase catalyses a reaction of adenosine to form inosine, said nucleoside phosphorylase catalysed a reaction of inosine to form hypoxanthine and said xanthine oxidase catalyses a reaction of said hypoxanthine to form urate and H 2 O 2 , and wherein an electrode in said biosensor detects said production of H 2 O 2 .

23 . A method according to any one of the preceding claims wherein detection of a marker is carried out using a biosensor that comprises Ruthenium Purple, optionally a biosensor that comprises an electrode and a layer of Ruthenium Purple on the electrode.

24 . A method of preparing an apparatus for measuring the level of at least one marker selected from glutamate, lactate, ATP, D-serine, acetylcholine and adenosine in whole blood, the method comprising;
(a) providing an apparatus capable of sampling, treating, taking measurements from or manipulating who

Assignees

• Ucl Business Plc

Inventors

• Ackland Gareth

Classifications

• G01N2800/24

  Immunology or allergic disorders · CPC title

• G01N2800/26

  Infectious diseases, e.g. generalised sepsis · CPC title

• G01N33/6812

  Assays for specific amino acids · CPC title

• G01N33/5047 Primary

  Cells of the immune system · CPC title

• C12Q1/005

  involving specific analytes or enzymes (including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence) · CPC title