Early Detection of Sepsis

• US 20080114576A1
• Filed: 12/09/2005
• Published: 05/15/2008
• Est. Priority Date: 12/09/2004
• Status: Abandoned Application

• Current Assignee: Secretary of State For Defence
• Original Assignee: Secretary of State For Defence
• Inventors: Brooks, Timothy John, Lukaszewski, Roman Antoni, Pearce, Martin Julian, Jackson, Matthew Christo, Yates, Amanda Marie
• Application Number:
  US11/792,724
• Publication Number:
  US 20080114576A1
• Time in Patent Office: Days
• US Class Current: 703/11
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 2800/26 Infectious diseases, e.g. generalised sepsis
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels
• CPC Class Codes: G16B 20/00 ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations
• CPC Class Codes: G16B 25/00 ICT specially adapted for hybridisation; ICT specially adapted for gene or protein expression
• CPC Class Codes: G16B 25/10 Gene or protein expression profiling; Expression-ratio estimation or normalisation
• CPC Class Codes: G16B 40/00 ICT specially adapted for biostatistics; ICT specially adapted for bioinformatics-related machine learning or data mining, e.g. knowledge discovery or pattern finding
• CPC Class Codes: G16B 40/10 Signal processing, e.g. from mass spectrometry [MS] or from PCR

Abstract

The invention describes a system and method for detecting early signs of infection and, in particular, for identifying individuals most likely to develop sepsis. Measurement of expression levels of particular combinations of cytokines and/or cellular activation markers, optionally combined with the use of predictive algorithms, allows a high degree of accuracy of prediction. The method is applicable in both civilian and military contexts.

Claims

1. Method for screening a biological sample to detect early stages of infection, SIRS or sepsis comprising the steps of:

a) detecting expression of a set of informative biomarkers by RT-PCR and/or detecting expression of a set of informative cell surface biomarkers by means of flow cytometry and/or monitoring a set of standard clinical measurements

b) analysing the results of detection by means of neural network or multivariate statistical analysis

c) classifying said sample according to the likelihood and timing of the development of overt infection.

2. Method according to claim 1, wherein analysis of the results yields a prediction of a probability of clinical SIRS or sepsis developing.

3. Method according to claim 1, wherein analysis of the results yields a binary yes/no prediction of clinical SIRS or sepsis developing.

4. Method according to claim 2, wherein if the development of clinical SIRS or sepsis is predicted, the results are subjected to a second analysis to determine the likely timing and/or severity of the clinical disease.

5. Method according to claim 1 wherein the set of informative biomarkers the expression of which is detected by means of RT-PCR consists of at least 6 selected from the list consisting of FasL, MCP-1, TNFα

, IL-1β

, IL-6, IL-8, IL-10, INF-α and

INF-γ

6. Method of claim 5 wherein the expression of at least 7 biomarkers is detected.

7. Method according to claim 1, wherein the set of informative cell surface biomarkers the expression of which is detected by means of flow cytometry consists of at least two selected from the list consisting of CD31, HLA-DR, CD54, CD11b, CD62L, CD 25, CD69 and CD80 and CD97

8. Method according to claim 1, wherein the standard clinical measurements analysed are selected from the list consisting of temperature, heart rate, total and differential white blood cell count (monocytes, lymphocytes, granulocytes, neutrophils), platelet count, serum creatinine, urea, lactate, base excess, pO₂, HCO₃⁻

, and C-reactive protein.

9. Method of claim 1 wherein analysis is by means of multivariate statistical analysis comprising principle component analysis and/or discriminant function analysis.

10. Method of claim 9 wherein the multivariate statistical analysis comprises discriminant function analysis

11. Method of claim 1 wherein analysis is by means of neural network.

12. Method of claim 10 wherein the neural network is a multilayered perception neural network.

13. Method of claim 9, wherein the analysis is capable of correctly predicting clinical SIRS or sepsis in greater than 80% of cases.

16. System according to claim 13 comprising means of performing both RT-PCR and FACS analysis.

17. System according to claim 13 further comprising means to acquire and analyse data from standard clinical measurements.

18. Analysis according to the method for screening a biological sample to detect early stages of infection, SIRS or sepsis comprising the steps of:

a) detecting expression of a set of informative biomarkers by RT-PCR and/or detecting expression of a set of informative cell surface biomarkers by means of flow cytometry and/or monitoring a set of standard clinical measurements

b) analysing the results of detection by means of neural network or multivariate statistical analysis

c) classifying said sample according to the likelihood and timing of the development of overt infection or using the system of claim 13 for the preparation of a diagnostic means for the diagnosis of infection, SIRS or sepsis.

19. Use of a method for screening a biological sample to detect early stages of infection, SIRS or sepsis comprising the steps of:

a) detecting expression of a set of informative biomarkers by RT-PCR and/or detecting expression of a set of informative cell surface biomarkers by means of flow cytometry and/or monitoring a set of standard clinical measurements

b) analysing the results of detection by means of neural network or multivariate statistical analysis

c) classifying said sample according to the likelihood and timing of the development of overt infection or the system according to claim 13 for the preparation of a diagnostic means for the diagnosis of infection, SIRS or sepsis.

14. System for screening a biological sample to detect early stages of infection, SIRS or sepsis comprising:

a means of extracting and purifying RNA from cells in said sample, a thermal cycler or other means to amplify selected RNA sequences by means of reverse transcription polymerase chain reaction (RT-PCR), a means of detecting and quantifying the results of said RT-PCR, a computer-based neural network trained so as to be able to analyse such results and a display means whereby the conclusion of the neural network analysis may be communicated to an operator.

15. System for screening a biological sample to detect early stages of SIRS or sepsis comprising:

a means of labelling specific cell surface markers and quantifying expression of said markers, a means of detecting and quantifying the results of said labelling and quantification, a computer-based neural network trained so as to be able to analyse such results and a display means whereby the conclusion of the neural network analysis may be communicated to an operator. Preferably, the labelling is by means of labelled antibodies or antibody fragments and the quantification is by means of fluorescence-activated cell sorting (FACS) or other form of flow cytometry.