Process for diagnosing chronic inflammatory intestinal diseases

Abstract

Some embodiments are directed to a process for quantifying changes in the intestinal mucosa caused by a chronic inflammatory intestinal disease in individuals, an ex vivo process for diagnosing a chronic inflammatory intestinal disease in individuals, and an ex vivo process for the differential diagnosis of Crohn's disease versus ulcerative colitis in individuals.

Claims

1. An ex vivo process for quantifying the modifications of the intestinal mucosa induced by a chronic inflammatory bowel disease in an individual, the process comprising: 1) Measuring, on a confocal endomicroscopic image of a digestive mucosa of an individual, following crypt well profile (PPC) parameters: a) Fluorescence leakage of the mucosa (FL), b) Perimeter of opening of the crypt well (PO), c) Sphericity of the crypt well (Spher), d) Roundness of the crypt well (Round), e) Feret diameter of the crypt well (Feret), f) Crypt well elongation factor (EF), g) Ratio of axes of opening of the crypt well (Ma/ma), h) Crypt density per field (Dens); and 2) quantifying modifications of the parameters a) to h) compared with corresponding parameters of a healthy individual.

2. An ex vivo process for diagnosing a chronic inflammatory bowel disease in an individual, the process comprising: 1) Measuring, on a confocal endomicroscopic image of digestive mucosa of an individual, following parameters: a) Fluorescence leakage of the mucosa (FL), b) Perimeter of opening of the crypt well (PO), c) Sphericity of a crypt well (Spher), d) Roundness of the crypt well (Round), e) Feret diameter of the crypt well (Feret), f) Crypt well elongation factor (EF), g) Ratio of axes of opening of the crypt well (Ma/ma), h) Crypt density per field (Dens); 2) Selecting values described as “higher extremes”, above thresholds defined below for each parameter: PO>474.62±103.10 μm, Spher>98.96±0.56%, Round>68.43±14.41%, Feret>169.88±51.08 μm, EF>1.745±0.266, Ma/ma>1.653±0.247, Dens>0.2837±0.0505; 3) Comparing, for each parameter, the mean of the “higher extreme” values and the mean value of FL to a pathological threshold, the pathological threshold being defined, for each parameter, in a following way: FL>10.0±3.6 LUT.sec.sup.−1, PO>666.40±103.10 μm, Spher>99.32±0.56%, Round>73.00±14.41%, Feret>186.11±51.08 μm, EF>1.87±0.266, Ma/ma>1.90±0.247, Dens>0.39±0.0505; 4) Assigning following scores: if the mean of the “higher extreme” values of each parameter PO, Spher, Round, Feret, EF, Ma/ma and Dens, or the mean value of FL, is less than the pathological threshold, the score of 0 is assigned to a corresponding parameter, if the mean of the “higher extreme” values of each parameter PO, Spher, Round, Feret, EF, Ma/ma and Dens, or the mean value of FL, is greater than the pathological threshold, a score of 1 is assigned for FL, PO, Round, Feret and Dens, and a score of 2 is assigned for Spher, EF and Ma/ma, and 5) Multiplying the sum of the scores by the sum of the number of values of each parameter, in which a total greater than 204±110 defines the individual as suffering from a chronic inflammatory bowel disease.

3. An ex vivo process for a differential diagnosis of Crohn's disease versus ulcerative colitis in an individual, the process comprising: the process defined in claim 2, followed, in the individual defined as suffering from an IBD, by a comparison of the mean value of FL and of the mean of the “higher extreme” values of the PO and Feret parameters, to a differential threshold defined in the following way: FL>19.86±5.70 LUT.sec.sup.−1, PO>618.32±98.95 μm and Feret>204.68±29.65 μm, in which the individual is defined as suffering from ulcerative colitis if, for each of the parameters FL, PO and Feret, a value greater than the corresponding differential threshold is reached, or the individual is defined as suffering from Crohn's disease in an opposite case.

4. The process as claimed in claim 1, wherein the total number of crypts analyzed in the endomicroscopic image of the digestive mucosa is greater than 20.

5. The process as claimed in claim 1, wherein the steps of measuring, of selecting values, of comparing and of assigning scores are automated by a computer program, stored on a non-statutory computer readable medium, for an a posteriori analysis of the endomicroscopy images or by a solution integrated into existing endomicroscopy software for a real-time analysis.

6. A computer program product, stored on a non-statutory computer readable medium, which can be downloaded from a communication network and/or recorded on a medium which can be read by computer and/or which can be executed by a processor, comprising: program code instructions for carrying out the process as claimed in claim 1, the computer program being executed on a computer or a processor.

7. A system, comprising: an endomicroscope, a visualizer, and a computer comprising image analysis means for measuring, for carrying out the process of claim 1.

8. The system as claimed in claim 7, wherein the endomicroscope is a confocal endomicroscope connected to a visualizer and a computer, the computer comprising an image analyzer for measuring, on a confocal endomicroscopic image of the digestive mucosa of an individual, parameters of the profiles of the crypt wells (PPC) chosen from FL, PO, Spher, Round, Feret, EF, Ma/ma and Dens, and a comparator suitable for comparing the measurements carried out with the corresponding values of a healthy individual.

9. The system as claimed in claim 8, wherein the image analysis means is suitable for selecting values described as “higher extreme”, greater than thresholds defined below for each parameter: PO>474.62±103.10 μM, Spher>98.96±0.56%, Round>68.43±14.41%, Feret>169.88±51.08 μm, EF>1.745±0.266, Ma/ma>1.653±0.247, Dens>0.2837±0.0505; and the comparator is suitable for comparing, for each parameter, the mean of the “higher extreme” values and the mean value of FL to a pathological threshold defined, for each parameter, in a following way: FL>10.0±3.6 LUT.sec.sup.−1, PO>666.40±103.10 μm, Spher>99.32±0.56%, Round>73.00±14.41%, Feret>186.11±51.08 μm, EF>1.87±0.266, Ma/ma>1.90±0.247, Dens>0.39±0.0505; the computer also including a score generator connected to the comparator and suitable for assigning following scores: if the mean of the “higher extreme” values of the PO, Spher, Round, Feret, EF, Ma/ma or Dens parameter, or the mean value of FL, is less than the threshold, a score of 0 is assigned to the corresponding parameter, if the mean of the “higher extreme” values of the PO, Spher, Round, Feret, EF, Ma/ma or Dens parameter, or the mean value of FL, is greater than the threshold, a score of 1 is assigned for FL, PO, Round, Feret and Dens and a score of 2 is assigned for Spher, EF and Ma/ma, and the computer also comprising a multiplier suitable for multiplying the sum of the scores by the sum of the number of values of each parameter.

10. The system as claimed in claim 9, wherein the comparator is suitable for comparing the mean value of FL and the mean of the “higher extreme” values of the PO and Feret parameters to a differential threshold defined in the following way: FL>19.86±5.70 LUT.sec.sup.−1, PO>618.32±98.95 μm and Feret>204.68±29.65 μm.

11. The process as claimed in claim 2, wherein the total number of crypts analyzed in the endomicroscopic image of the digestive mucosa is greater than 20.

12. The process as claimed in claim 3, wherein the total number of crypts analyzed in the endomicroscopic image of the digestive mucosa is greater than 20.

13. The process as claimed in claim 2, wherein the steps of measuring, of selecting the values, of comparing and of assigning the scores are automated by a computer program, stored on a non-statutory computer readable medium, for an a posteriori analysis of the endomicroscopy images or by a solution integrated into the existing endomicroscopy software for a real-time analysis.

14. The process as claimed in claim 3, wherein the steps of measuring, of selecting the values, of comparing and of assigning the scores are automated by a computer program, stored on a non-statutory computer readable medium, for an a posteriori analysis of the endomicroscopy images or by a solution integrated into the existing endomicroscopy software for a real-time analysis.

15. The process as claimed in claim 4, wherein the steps of measuring, of selecting values, of comparing and of assigning scores are automated by a computer program, stored on a non-statutory computer readable medium, for an a posteriori analysis of the endomicroscopy images or by a solution integrated into existing endomicroscopy software for a real-time analysis.

16. A computer program product, stored on a non-statutory computer readable medium, which can be downloaded from a communication network and/or recorded on a medium which can be read by computer and/or which can be executed by a processor, comprising: program code instructions for carrying out the process as claimed in claim 2, the computer program being executed on a computer or a processor.

17. A computer program product, stored on a non-statutory computer readable medium, which can be downloaded from a communication network and/or recorded on a medium which can be read by computer and/or which can be executed by a processor, comprising: program code instructions for carrying out the process as claimed in claim 3, the computer program being executed on a computer or a processor.

18. A system, comprising: an endomicroscope, a visualizer, and a calculator, for carrying out the process of claim 2.

19. A system, comprising: an endomicroscope, a visualizer, and a calculator, for carrying out the process of claim 3.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1 represents the scheme of the principle of laser confocal endomicroscopy. The vertical rectangle at the top represents the source of laser light and the detector of the confocal system. The horizontal rectangle represents the pinhole through which light rays pass. The horizontal oval represents a focusing lens. The laser radiation passes through the center of the lens and reaches the tissue. The fluorescent molecule present in the tissue then emits light upwards. The light rays outside the focal plane are diverged by the lens and do not pass through the pinhole. The light rays emitted by the focal plane are converged by the lens toward the pinhole and the detector.

(2) FIG. 2 represents the considered architectural parameters of the crypts. FIG. 2A represents the perimeter of opening of the crypt. FIG. 2B represents the elongation factor. FIG. 2C represents the sphericity. FIG. 2D represents the Ma/ma ratio of the crypt opening. FIG. 2E represents the roundness. FIG. 2F represents the Feret diameter. FIG. 2G represents the crypt density per field.

(3) FIG. 3 represents the comparison of the mean of the values greater than the 90.sup.th percentile of the healthy group, for the following parameters: A: perimeter of opening of the crypt (expressed in μm), B: Sphericity (expressed as %), C: Roundness (expressed as %), D: Feret diameter (expressed in μm), E: Elongation factor, F: Ma/ma and G: Density. Each of these parameters is evaluated in 5 healthy individuals (“S”), 10 individuals suffering from Crohn's disease (“CD”) and 11 individuals suffering from ulcerative colitis (“UC”).

(4) FIG. 4 represents the comparison of the mean of the fluorescence leakage (FL) values, expressed in LUT.sec.sup.−1, in 5 healthy individuals (“S”), 10 individuals suffering from Crohn's disease (“CD”) and 11 individuals suffering from ulcerative colitis (“UC”).

EXAMPLES

Example 1: Validation of Diagnostic Imaging Biomarkers for Chronic Inflammatory Bowel Diseases by Confocal Endomicroscopy

(5) Materials and Methods

(6) 5 healthy patients, 10 patients suffering from Crohn's disease (CD) in clinical inflammatory remission phase (CDAI<150) and 11 patients suffering from ulcerative colitis (UC) in clinical inflammatory remission phase (Mayo score <1) were included in this retrospective study.

(7) The fluorescence leakage (FL) in the colonic mucosa, the perimeter of opening (PO), the roundness (Round), the sphericity (Spher), the elongation factor (EF), the Feret diameter (Feret), the large axis to small axis ratio (Ma/ma) and the crypt well density (Dens) were quantified by measuring or calculating these parameters by the confocal endomicroscopy images (ColoFlex UHD, Cellvizio, Mauna Kea Technologies).

(8) Only the values higher than the 90.sup.th centile of the healthy patients were considered for the analysis (values described as “higher extremes”).

(9) Results

(10) The quantitative analysis was carried out on 91±8 crypts per patient. It was possible to demonstrate that the mean of the values higher than the 90.sup.th centile of the healthy patients for the sphericity (Spher) (p<0.01), roundness (Round) (p<0.05), Feret diameter (Feret) (p<0.05), elongation factor (EF) (p<0.01), and Ma/ma ratio (p<0.01) are significantly higher for the CD group compared with the healthy patients (cf. FIG. 3).

(11) The mean of the values higher than the 90.sup.th centile of the healthy patients for the perimeter of opening (PO) (p<0.05), sphericity (Spher) (p<0.01), Feret diameter (Feret) (p<0.01), elongation factor (EF) (p<0.01), Ma/ma ratio (p<0.01) and crypt image density (Dens) (p<0.05) are significantly higher for the UC group compared with the healthy patients (cf. FIG. 3).

(12) The mean of the values higher than the 90.sup.th centile of the healthy patients for the perimeter of opening (PO) (p<0.05) and Feret diameter (Feret) (p<0.05) are significantly higher for the UC group compared with the CD group (cf. FIG. 3).

(13) Finally, 5.9 times more fluorescein leakage (FL) in the colonic mucosa was observed in the UC group compared with the healthy group (p<0.05) and 2.9 times more was observed compared with the CD group (p<0.05) (cf. FIG. 4).

(14) Conclusion

(15) This study demonstrates that it is possible to quantify the architectural modifications of the mucosa induced by IBDs, even during a clinical remission. It was possible to define 3 types of parameter: 1/ the parameters common to IBDs (sphericity, Feret diameter, elongation factor, Ma/ma); 2/ a parameter specific for Crohn's disease (Roundness) and 3/ parameters specific for ulcerative colitis (fluorescence leakage, perimeter of opening and density).

(16) These results set down the bases for establishing an optical score for IBDs which it will be possible to use for diagnosis and monitoring of the therapeutic response.

(17) These results also validate the use of diagnostic imaging biomarkers for IBDs and specifically for CD and UC.

Example 2: Validation of a Diagnostic Score by Endomicroscopic Imaging of IBDs, of CD and of UC

(18) Materials and Methods

(19) 5 healthy patients, 10 patients suffering from Crohn's disease (CD) in clinical inflammatory remission phase (CDAI<150) and 11 patients suffering form ulcerative colitis (UC) in clinical inflammatory remission phase (Mayo score <1) were included in this retrospective study. The fluorescence leakage (FL) in the colonic mucosa, the perimeter of opening (PO), the roundness (Round), the sphericity (Spher), the elongation factor (EF), the Feret diameter (Feret), the large axis to small axis ratio (Ma/ma) and the crypt well density (Dens) were quantified by measuring or calculating these parameters by the confocal endomicroscopy images (ColoFlex UHD, Cellvizio, Mauna Kea Technologies).

(20) Only the values higher than the 90th centile of the healthy patients were considered for the analysis (values described as “higher extremes”).

(21) The diagnostic score for IBDs is defined by:

(22) 1—Selection of the “higher extreme” values, that is to say the values higher than the 90.sup.th centile with the healthy patients for each of the parameters PO, Spher, Round, Feret, EF, Ma/ma and Dens.

(23) The value of the 90.sup.th centile of the healthy patients for each of the parameters is measured/calculated, with the following results: PO>474.62±103.10 μm, Spher>98.96±0.56%, Round>68.43±14.41%, Feret>169.88±51.08 μm, EF>1.745±0.266, Ma/ma>1.653±0.247, Dens>0.2837±0.0505;

(24) 2—For each of these parameters, the mean of the “higher extreme” values, that is to say higher than the 90.sup.th centile of the healthy patients, and the mean value of FL are compared to a pathological threshold defined by a consensus of experts. The pathological threshold, for each parameter, is thus defined by the following values: FL>10 LUT.sec.sup.−1, PO>666.40 μm, Spher>99.32%, Round>73.00%, Feret>186.11 μm, EF>1.87, Ma/ma>1.90, Dens>0.39.

(25) If the mean of the “higher extreme” values of the PO, Spher, Round, Feret, EF, Ma/ma or Dens parameter, or the mean value of FL, is less than the pathological threshold, the score of 0 is assigned to the parameter. If this value is greater than the pathological threshold, the score of 1 is assigned for FL, PO, Round, Feret and Dens and the score of 2 is assigned for Spher, EF and Ma/ma.

(26) 3—The sum of the scores is multiplied by the sum of the number of values of each parameter. In this example, a total greater than 204 defines the patient as suffering from an IBD.

(27) 4—For the patients defined IBD, the mean value of FL and the mean of the “higher extreme” values of PO and Feret are compared to a differential threshold defined by a consensus of experts. The differential threshold, for each of these parameters, is thus defined by the following values: FL>19.86 LUT.sec.sup.−1, PO>618.32 μm Feret>618.32 μm.

(28) If, for each of these 3 parameters, a value greater than the differential threshold corresponding to the parameter in question is measured/calculated, then the patient is defined as suffering from UC. In the opposite case, that is to say if at least one of the parameters does not have a value greater than the differential threshold of the corresponding parameter, then the patient is defined as suffering from CD.

(29) Results

(30) The values of the 90.sup.th centile of the healthy population are: Perimeter of opening (>474.62 μm), Sphericity (>98.96%), Roundness (>68.43%), Feret diameter (>169.88 μm), Elongation factor (>1.745), Ma/ma (>1.653) and Density (>0.2837).

(31) The values, occurrences, scores and diagnoses of IBD are summarized in table 1. The specificity, the sensitivity, the positive predictive value (PPV) and the negative predictive value (NPV) are 100% for this group of patients.

(32) TABLE-US-00001 TABLE 1 Perimeter of opening Sphericity Roundness Feret diameter Occur- Occur- Occur- Occur- Patient Mean Point rence Mean Point rence Mean Point rence Mean Point rence 1 healthy 564.06 0 7 99.31 0 5 70.35 0 9 0 0 0 2 healthy 603.092 0 4 99.26 0 3 72.33 0 4 0 0 0 3 healthy 737.324 1 4 99.19 0 2 75.6 1 2 216.3896 1 8 4 healthy 515.508 0 3 99.21 0 2 0 0 0 175.9772 0 9 5 healthy 525.98 0 1 99.31 0 7 69.02 0 4 185.9256 0 2 6 IBD CD 584.052 0 6 99.6 2 21 73.36 1 24 201.11 1 7 7 IBD CD 612.612 0 9 99.53 2 7 73.12 1 10 216.342 1 6 8 IBD CD 642.124 0 44 99.74 2 16 74.67 1 24 237.3336 1 56 9 IBD CD 544.544 0 17 99.27 0 3 75.62 1 10 206.4888 1 28 10 IBD CD 667.828 1 31 99.68 2 32 73.64 1 37 235.1916 1 47 11 IBD CD 825.384 1 6 99.85 2 27 76 1 29 256.1356 1 10 12 IBD CD 751.128 1 31 99.72 2 6 74.2 1 7 249.8524 1 41 13 IBD CD 695.436 1 19 99.65 2 24 74 1 19 238.714 1 34 14 IBD CD 490.756 0 1 99.73 2 13 73.54 1 24 186.6396 1 6 15 IBD CD 593.096 0 8 99.79 2 13 74.43 1 20 202.9188 1 20 16 IBD CD 754.936 1 21 99.57 2 7 75.29 1 8 247.996 1 30 17 IBD UC 857.752 1 11 99.8 2 54 74.61 1 57 252.518 1 18 18 IBD UC 766.836 1 35 99.7 2 20 73.14 1 19 260.6576 1 43 19 IBD UC 689.248 1 10 99.64 2 17 74.05 1 30 219.3408 1 15 20 IBD UC 904.4 1 16 99.61 2 36 75.22 1 40 313.3984 1 16 21 IBD UC 731.136 1 3 99.76 2 40 73.45 1 39 276.7464 1 4 22 IBD UC 601.664 0 5 99.51 2 9 71.72 0 29 212.8672 1 8 23 IBD UC 666.4 0 13 99.61 2 39 73.67 1 60 246.7584 1 15 24 IBD UC 838.236 1 3 99.47 2 12 74.86 1 20 243.5692 1 6 25 IBD UC 753.508 1 35 99.84 2 6 71.34 0 10 282.744 1 37 26 IBD UC 788.256 1 31 99.68 2 9 71.64 0 9 285.4572 1 34 Elongation factor Ma/ma Density Fluorescence leakage Occur- Occur- Occur- Occur- IBD diagnostic score Mean Point rence Mean Point rence Mean Point rence Mean Point rence (retrospective) 0 0 0 0 0 0 0.3672 0 3 11.15 1 NA 24 healthy 1.869 0 10 1.784 0 10 0.3426 0 4 5.1 0 NA 0 healthy 2.021 2 6 1.884 0 7 0.4695 1 5 2.08 0 NA 204 healthy 1.843 0 3 1.712 0 2 0 0 0 3.77 0 NA 0 healthy 0 0 0 0 0 0 0.3114 0 1 8.26 0 NA 0 healthy 2.092 2 16 2.121 2 9 0.4217 1 15 12.14 1 NA 980 IBD 1.968 2 3 1.787 0 3 0.3645 0 7 19.87 1 NA 315 IBD 2.426 2 43 2.015 2 22 0.4429 1 47 10.21 1 NA 2520 IBD 2.074 2 24 1.865 0 8 0.5579 1 29 12.4 1 NA 714 IBD 2.536 2 42 2.074 2 28 0.4522 1 48 6.835 0 NA 2650 IBD 2.146 2 8 1.814 0 4 0.4202 1 43 11.97 1 NA 1143 IBD 2.493 2 22 2.061 2 10 0.3713 0 15 14.92 1 NA 1320 IBD 2.4 2 38 1.993 2 25 0.4494 1 40 1.32 0 NA 1990 IBD 1.995 2 17 1.915 2 9 0.3983 1 17 7.64 0 NA 783 IBD 2.284 2 28 2.045 2 11 0.3956 1 22 23.68 1 NA 1220 IBD 2.313 2 20 2.109 2 15 0.4855 1 32 19.87 1 NA 1463 IBD 1.976 2 24 1.898 0 18 0.4375 1 37 28.87 1 NA 1971 IBD 2.347 2 34 2.164 2 20 0.5083 1 44 9.81 0 NA 2150 IBD 2.389 2 24 1.973 2 18 0.4977 1 37 23.82 1 NA 1661 IBD 2.39 2 12 1.967 2 5 0.5951 1 17 70.32 1 NA 1562 IBD 2.539 2 4 1.914 2 2 0.3929 1 16 37.88 1 NA 1188 IBD 2.358 2 8 1.927 2 6 0.4042 1 17 50.59 1 NA 738 IBD 2.147 2 19 1.921 2 9 0.4305 1 26 23.79 1 NA 1810 IBD 1.893 2 4 1.675 0 2 0.4 1 23 50.41 1 NA 630 IBD 2.681 2 28 2.271 2 17 0.5042 1 45 58.6 1 NA 1780 IBD 2.461 2 29 2.065 2 16 0.5632 1 37 23.15 1 NA 1650 IBD

(33) The values and differential diagnoses are summarized in table 2.

(34) For the CD diagnosis, the specificity is 87.5%, the sensitivity is 100%, the PPV is 83.3% and the NPV is 100%.

(35) For the UC diagnosis, the specificity is 100%, the sensitivity is 81.8%, the PPV is 100% and the NPV is 88.2%.

(36) TABLE-US-00002 TABLE 2 IBD diagnostic Perimeter score of Feret Fluorescence Differential Patient (retrospective) opening diameter leakage diagnosis 6 IBD CD 980 IBD 584.052 201.11 12.14 CD 7 IBD CD 315 IBD 612.612 216.342 19.87 CD 8 IBD CD 2520 IBD 642.124 237.3336 10.21 CD 9 IBD CD 714 IBD 544.544 206.4888 12.4 CD 10 IBD CD 2650 IBD 667.828 235.1916 6.835 CD 11 IBD CD 1143 IBD 825.384 256.1356 11.97 CD 12 IBD CD 1320 IBD 751.128 249.8524 14.92 CD 13 IBD CD 1990 IBD 695.436 238.714 1.32 CD 14 IBD CD 783 IBD 490.756 186.6396 7.64 CD 15 IBD CD 1220 IBD 593.096 202.9188 23.68 CD 16 IBD CD 1463 IBD 754.936 247.996 19.87 UC 17 IBD UC 1971 IBD 857.752 252.518 28.87 UC 18 IBD UC 2150 IBD 766.836 260.6576 9.81 CD 19 IBD UC 1661 IBD 689.248 219.3408 23.82 UC 20 IBD UC 1562 IBD 904.4 313.3984 70.32 UC 21 IBD UC 1188 IBD 731.136 276.7464 37.88 UC 22 IBD UC 738 IBD 601.664 212.8672 50.59 CD 23 IBD UC 1810 IBD 666.4 246.7584 23.79 UC 24 IBD UC 630 IBD 838.236 243.5692 50.41 UC 25 IBD UC 1780 IBD 753.508 282.744 58.6 UC 26 IBD UC 1650 IBD 788.256 285.4572 23.15 UC

(37) Conclusion

(38) This study demonstrates that the endomicroscopic score established enables the diagnosis of an IBD, even in a state of clinical inflammatory remission. This score coupled to the use of 3 other biomarkers (FL, PO and Feret diameter) also makes it possible to distinguish between CD and UC.

Example 3: Validation of a Diagnostic Score by Endomicroscopic Imaging for IBDs (CD and UC)

(39) Materials and Methods

(40) 4 healthy patients, 13 patients suffering from Crohn's disease (CD) in clinical inflammatory remission phase (CDAI<150) and 16 patients suffering from ulcerative colitis (UC) in clinical inflammatory remission phase (Mayo score <1) were included in this retrospective study. The parameters described in example 2 were quantified by measuring or calculating these parameters by the confocal endomicroscopy images (ColoFlex UHD, Cellvizio, Mauna Kea Technologies).

(41) Only the values higher than the 90.sup.th centile of the healthy patients were considered for the analysis (values described as “higher extreme”). The diagnostic score is constructed as previously described. The differential diagnosis is constructed on the basis of FL (>19.86±5.70 LUT.sec.sup.−1), PO (>618.32±98.95 μm) and Feret (>204.68±29.65 μm) as previously described.

(42) Results

(43) The results are summarized in table 3.

(44) TABLE-US-00003 TABLE 3 Patient Diagnosis Retrospective No. at diagnosis Differential diagnosis series 2 inclusion Score Healthy/IBD FL PO Feret CD/UC 1 Healthy 1 Healthy 2 Healthy 172 Healthy 3 Healthy 0 Healthy 4 Healthy 296 IBD 5 CD 720 IBD 20.87 744.75 367.26 UC 6 CD 456 IBD 23.185 557.93 206.21 CD 7 CD 1211 IBD 9.9 847.17 296.94 CD 8 CD 2000 IBD 11.21 728.88 262.29 CD 9 CD 1152 IBD 13.95 859.54 311.17 CD 10 CD 627 IBD 6.06 677.67 238.51 CD 11 CD 1184 IBD 22.46 697.33 251.08 UC 12 CD 1140 IBD 34.18 708.28 254.43 UC 13 CD 2112 IBD 33.41 553.52 201.3 CD 14 CD 62 Healthy 3.89 0 0 CD 15 CD 3374 IBD 8.16 875.5 314.47 CD 16 CD 1904 IBD 16.56 889.23 328.48 UC 17 CD 1112 IBD 19.49 859.54 311.17 UC 18 UC 252 IBD 7.326 647.17 225.6 CD 19 UC 3724 IBD 26.14 697.06 240.68 UC 20 UC 1435 IBD 19.4675 741.9 258.98 UC 21 UC 3530 IBD 29.96 664.13 246.18 UC 22 UC 1425 IBD 7.326667 719.18 264.71 CD 23 UC 425 IBD 22.614 647.28 232.59 UC 24 UC 930 IBD 44.374 707.06 242.93 UC 25 UC 3059 IBD 37.88 605.3 215.47 CD 26 UC 1362 IBD 18.784 736.61 242.4 UC 27 UC 748 IBD 12.6 690.25 236.94 CD 28 UC 1350 IBD 23.84 689.14 238.69 UC 29 UC 355 IBD 3.416667 655.85 223.13 CD 30 UC 258 IBD 14.52 615.79 215.41 CD 31 UC 978 IBD 19.30667 664.56 231.84 UC 32 UC 3240 IBD 7.43 758.54 275.53 CD 33 US 1115 IBD 21.935 696.47 242.17 UC

(45) The sensitivity of the IBD diagnosis is 96.55% for a specificity of 75%. The sensitivity of the UC/CD differential diagnosis is 68.75% for a specificity of 61.54%.

(46) Conclusion

(47) This study demonstrates the validity of the endomicroscopic score established and enables the diagnosis of an IBD, even in clinical inflammatory remission state. This score coupled to the use of 3 other biomarkers (FL, PO and Feret) also makes it possible to distinguish between CD and UC.

(48) The whole of the population studied (example 2 and example 3) demonstrates that the diagnostic score enables the identification of an IBD with a sensitivity of 98% and a specificity of 89%. The differential diagnosis has a sensitivity of 65.4% and a specificity of 75%.

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