Device and method for detecting biomarkers in stoma dejecta and model dejecta

Abstract

A device and method for detecting a presence and / or concentrations level of a nutrient or biomarker in a stoma dejecta sample is disclosed herein. A sample pad is disposed on a backing layer, a conjugate release pad is in fluid communication with the sample pad, a first material is disposed in the conjugate release pad, and a test line is in fluid communication with the conjugate release pad. The first material is disposed in the conjugate release pad and is selected in accordance with the nutrient or biomarker. The sample deposited on the sample pad flows through the conjugate release pad so that the nutrient or biomarker interacts with the first material to form a second material that flows to the test line. The test line is configured to change color in response accumulation of the second material thereon. A model dejecta for optimizing and calibrating the device is also disclosed herein.

Description

DEVICE AND METHOD FOR DETECTINGBIOMARKERS IN STOMA DEJECTA AND MODEL DEJECTACROSS-REFERENCE TO RELATED APPLICATION

[0001] The present application claims the benefit of priority to Bellamy et al., U.S. Provisional Patent Application Serial No. 63 / 562,058, filed March 6, 2024, and entitled “System and Method for Detecting Biomarkers in Stoma Dejecta”, the entire contents of which are incorporated herein by reference.FIELD OF DISCLOSURE

[0002] The present subject matter relates to detection of biomarkers and more particularly, a device and method for detecting biomarkers in stoma dejecta.BACKGROUND

[0003] Ostomy procedures are surgeries that redirect the flow of gastrointestinal or bladder contents to avoid diseased, damaged, or missing organs of a patient. One end of an organ is connected to the skin of a patient (“ostomate”) through the abdominal wall bypassing the diseased or excised organ, creating a stoma. The most common types of ostomy procedures include ileostomy, colostomy, and urostomy. In an ileostomy, the ileum of the small intestine forms the stoma, bypassing the whole large intestine. In a colostomy, the colon of the large intestine forms the stoma, bypassing lower portions of the large intestine such as the lower colon or the rectum. In a urostomy, the ureter is surgically attached to a section of the ileum, which is then exteriorized percutaneously to form the stoma, or the ureter can form the stoma. Both methods bypass the bladder and enable drainage of urine.

[0004] Ostomy procedures are most commonly performed for people with birth defects, cancer, Crohn’s disease, inflammatory bowel disease (TBD), diverticulitis, incontinence, severe abdominal trauma, and the like. Ostomies can be permanent or temporary, requiring another surgery to later close the stoma and reconnect the organs in the bowel. Ostomies do not lower life expectancy, but they can lower patient quality of life due to complications at the stoma intestine-skin interface or due to inconveniences associated with the ostomy pouch or adhesive barrier, such as peristomal skin complications.

[0005] An ostomy system may include an ostomy pouch and a barrier appliance comprising a skin barrier and / or ostomy wafer for attaching the pouch to the stoma of the patient. For ostomates whose anuses have been bypassed, intestinal contents, also called dejecta, may be collected in the ostomy pouches attached to the stoma instead of leaving the body through the anus. Ostomy pouches may hold between 400 to 2000 mb of dejecta depending on size. Ostomates may choose the ostomy pouch size based on convenience with their clothing and activities and how long they expect to wait before changing a pouch. Ostomy pouches typically fall into two categories: closed- end pouches and drainable pouches. Closed-end pouches are non-drainable, requiring replacement approximately twice a day, and are best suited for patients with soft to well-formed stoma output, typically colostomy patients. In contrast, drainable pouches may come with an opening at the bottom, allowing for the contents to be emptied as needed. Open pouches may permit the dejecta to be emptied directly into the toilet, usually at least twice a day.

[0006] In addition, ostomates may use a one-piece or a two-piece ostomy system. A one-piece ostomy system may include a pouch that is integrated with the barrier appliance into a unified unit. In such system, both the pouch and the barrier appliance may be detached from the ostomate and replaced as a unit. Alternately, a two-piece system may allow the ostomy pouch to be separated from the barrier appliance, which remains attached to the ostomate. Thus, in a two-piece ostomy system, the pouch may be changed while barrier appliance may remain attached to the skin of the ostomate.

[0007] In general, ostomy dejecta may have a similar composition to feces of people without an ostomy, or may vary based on the anatomical location of the stoma. Non-ostomy feces are about 75% water by mass and also contain organic and inorganic solids. Dejecta from a high output ostomy may have a higher fluid proportion. In some cases, dejecta have been found to comprise about 90% water and varying amounts of sodium, potassium, and other ions. Inorganic and organic compounds found in dejecta may include electrolytes, bacterial biomass, carbohydrates, fiber, proteins, and fats. The specific compositions of the organic and inorganic solids depends on individual diet, exercise, environment, and water intake of the patient. Bacteria species in theorganic mass also vary between patients. Dejecta composition also may vary by individual lifestyle.

[0008] In addition to differences in lifestyle, different ostomates may have varying needs that will determine the placement of their ostomy. For example, different ostomates may have different amounts of their digestive or urinary tracts bypassed. In the case of a urostomy, the composition of the dejecta is not subject to change on the basis of bypassing various parts of the ureter or bladder which function to move, store, and release urine rather than absorb nutrients from it. In the cases of colostomy and ileostomy, however, dejecta composition may differ based on the bypassed sections of the small intestine and / or large intestine. As both the small and large intestines contribute to functions such as nutrient absorption and dehydration of stool. Missing parts of these organs can significantly change the composition of dejecta. For example, bypassing more than 50 cm of the ileum may lead to nutritional deficiencies. For ostomates with ileostomies that remove or bypass a large amount of the ileum, dejecta may contain more nutrients than people with no ileostomy or people with shorter lengths of ileum bypassed.SUMMARY

[0009] According to one aspect, a device adapted to detect a presence and / or concentration level of a nutrient and / or biomarker in a stoma dejecta sample includes a backing layer, a sample pad disposed on the backing layer, a conjugate release pad in fluid communication with the sample pad, a first material disposed in the conjugate release pad, and a test line in fluid communication with the conjugate release pad. The first material is selected in accordance with the nutrient or biomarker. The device is configured to cause the stoma dejecta sample deposited on the sample pad to flow through the conjugate release pad so that the nutrient or biomarker interacts with the first material to form a second material that flows to the test line, wherein the test line is configured to change color in response to accumulation of the second material thereon.

[0010] According to another aspect, a method of detecting presence and / or a concentration level of a nutrient or biomarker in a stoma dejecta sample using a biomarker testing device that includes a sample pad, a conjugate release pad having a first material disposed therein, and a test line includes depositing the stoma dejecta sample on the sample pad and causing the stoma dejectato flow through the conjugate release pad. The method further includes causing an interaction of the nutrient or biomarker with the first material to form a second material, wherein the first material is selected in accordance with the nutrient or biomarker, and causing the second material to accumulate at the test line, and thereby causing the test line to change color.

[0011] In some embodiments, the biomarker or nutrient includes glucose, alcohol, pyruvate oxidase, or creatinine. In such embodiments, the first material includes an enzyme. In some cases, the second material includes hydrogen peroxide.

[0012] In some embodiments, the biomarker includes a protein.

[0013] In some embodiments, the first material includes a nanoparticle.

[0014] In some embodiments, the biomarker is one or more of polymorphonuclear neutrophil elastase, lactoferrin, macrophage cytosolic protein S100A12 (calgranulin C), pyruvate kinase, interleukin 1, interleukin 6, interleukin 8, tumor necrosis factor a, glucose, myeloperoxidase, al- antitrypsin, neutrophil gelatinase-associated lipocalin, hemoglobin, or a matrix metalloprotease.

[0015] In some embodiments, the biomarker is one of inflammatory cytokines and chemokines.

[0016] In some embodiments, the device is a Lateral Flow Immunoassay (LFIA)

[0017] In some embodiments, the device is an Enzyme-based Barcode-style Lateral-flow Assay.

[0018] According to another aspect, a model dejecta composition for optimizing and calibrating the device includes between 70 and 90 percent water, the nutrient and / or biomarker, and one or more of carbohydrates, bacterial biomass, fat, proteins, and inorganics. The model dejecta composition has a viscosity of between approximately 3,500 and approximately 5,500 centipoise (cps) at 50 rpm.

[0019] In some embodiments, wherein the model dejecta composition is acidic.

[0020] In some embodiments, the model dejecta composition further comprises 6.8g NaCl, 6.8g KC1, 3.4g CaC12 per liter of water.

[0021] In some embodiments, the model dejecta composition further comprises 333g rice flour per liter of water.

[0022] In some embodiments, the model dejecta composition further comprises 100g polyethylene glycerol per liter of water.

[0023] In some embodiments, the model dejecta composition further comprises 5g collagen per liter of water or 10% cellulose per liter of water.

[0024] In some embodiments, the model dejecta composition further comprises 60 grams peanut oil per liter of water.

[0025] In some embodiments, the model dejecta composition further comprises 120g yeast per liter of water.

[0026] In some embodiments, the model dejecta composition further comprises a biomarker or a nutrient.

[0027] In some embodiments, an ostomy system includes an ostomy pouch and a barrier appliance and the device is incorporated into the ostomy system. In some cases, the device is disposed in the ostomy pouch. Further, in some cases, he the ostomy pouch includes a clear window and the device is viewable through the clear window from an exterior of the ostomy pouch.

[0028] In some embodiments, a point-of-care or at-home test kit includes the device.

[0029] In some embodiments, a computational model to predict a presence and / or concentration of a nutrient and / or biomarker in dejecta in accordance with a placement of a stoma on ostomate is configured using a plurality of the devices to analyze a component of dejecta associated with a corresponding plurality of stomas disposed in different locations.

[0030] In some embodiments, a computational model to determine a correlation between a presence and / or concentration of a nutrient and / or a biomarker and environmental and / or genetic factors is configured using a plurality of devices to analyze a component of dejecta from a plurality of ostomates associated with a plurality of environmental and / or genetic factors.

[0031] Other aspects and advantages will become apparent upon consideration of the following detailed description and the attached drawings wherein like numerals designate like structures throughout the specification.BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIG. 1 is an elevational view of a body side of a one-piece ostomy system with which the device of the present disclosure may be used;

[0033] FIG. 1 A is an elevational view of a pouch and a barrier appliance of a two-piece ostomy system with which the device of the present disclosure may be used;

[0034] FIG. 2 is a top plan view of a biomarker test device in accordance with the present disclosure;

[0035] FIG. 2A is a cross-sectional view taken along the line 2A-2A of FIG. 2; and

[0036] FIG. 3 is a plot showing a relationship between intensity of a test line and elapsed time associated with different model dejecta samples.DET AILED DESCRIPTION

[0037] While the present disclosure is susceptible of embodiment in various forms, there is shown in the drawings and will hereinafter be described presently preferred embodiments with the understanding that the present disclosure is to be considered an exemplification and is not intended to limit the disclosure to the specific embodiments illustrated. The words “a” or “an” are to be taken to include both the singular and the plural. Conversely, any reference to plural items shall, where appropriate, include the singular. The words “first,” “second,” “third,” and the like may be used in the present disclosure to describe various information, such information should not be limited to these words. These words are only used to distinguish one category of information from another. The directional words “top,” “bottom,” up,” “down,” front,” “back,” and the like are used for purposes of illustration and as such, are not limiting. Depending on the context, the word “if’ as used herein may be interpreted as “when” or “upon” or “in response to determining.”

[0038] Analyzing ostomy dejecta compositions may provide a number of insights. Firstly, the presence of certain nutrients and / or biomarkers at specific concentrations may be correlated with formation or progression of a disease phenotype. Further, monitoring components of dejecta such as glucose, elastase, fecal calprotectin (FCP), fecal lactoferrin (LF), macrophage cytosolic protein S100A1, pyruvate kinase, matrix metalloproteases, calgranulin C, and inflammatory cytokines andchemokines (for example, interleukin 1, interleukin 6, interleukin 8, and / or tumor necrosis factor alpha) levels may provide indications of progression, treatment, and / or mitigation of, for example, inflammation related to mucosal healing, inflammatory bowel disease (IBD), Crohn’s disease (CD) and ulcerative colitis, and the like, and may reduce need for repeated endoscopy procedures to monitor such conditions. In addition, the placement of the stoma in relation to the bowel may influence the waste contents by levels of water and bacteria in dejecta since the further along the bowels waste is allowed to move creates more nutrient absorption and interaction with the gut microbiome. For example, the densities and bacterial diversities vary in different regions of the gut - duodenum (103 / g), jejunum (104 / g), ileum (107 / g), and colon (1012bacteria / g).

[0039] The two main groups of ostomates who undergo ostomy procedures are people with IBD and people with colorectal cancer (CRC). Ostomates having such conditions have to undergo frequent procedures such as carcinoembryonic antigen tests, CT scans, endoscopies, and the like to monitor disease progression and treatment efficacy. For example, a patient treated for CRC may have to undergo such tests every 3-6 months after treatment. Likewise, patients with IBD may have to undergo endoscopies every 3-6 months after a change in medication to monitor mucosal healing, the primary goal in IBD treatment.

[0040] One or more biomarker detection devices configured to detect nutrients, biomarkers, and other analytes in dejecta may reduce the burden on ostomates if disease progression and treatment efficacy could be monitored without endoscopy and other types of testing procedures.

[0041] Currently, there is a limited body of research on the impacts of ostomy procedures, particularly regarding how stoma anatomical placement influences waste composition and its subsequent interplay with various physiological conditions. This gap in knowledge is significant considering the gastrointestinal tract’s integral connection not only to the immune and neural systems but also to overall metabolic processes. Analyzing dejecta composition may also provide a number of insights to diagnose, monitor, and / or treat disease and how stoma placement influences waste composition.

[0042] The presence of certain biomarkers at specific concentrations in dejecta may be correlate to the formation, progression, or differential diagnosis of a disease phenotype. The presence or levels of biomarkers such as fecal calprotectin (FC), fecal lactoferrin (LF), glucose, polymorphonuclear neutrophil (PMN)-elastase, macrophage cytosolic protein S100A12(calgranulin C), pyruvate kinase, matrix metalloproteases (MMPs), inflammatory cytokines and chemokines (for example, interleukin 1, interleukin 6, interleukin 8, and / or tumor necrosis factor alpha), myeloperoxidase, fecal clearance of plasma alpha 1 -antitrypsin, fecal neutrophil gelatinase- associated lipocalin (NGAL), and hemoglobin may be monitored to determine presence of inflammation related to cancer, IBD, and other inflammatory conditions.

[0043] FC is a calcium and zinc-binding protein with versatile functions, constituting 60% of neutrophil cytosolic protein. Functions include antibacterial and antifungal activity, inhibition of metalloproteinases, and induction of apoptosis. Concentrations of FC may be correlated with active inflammation in the gut, and the ability to distinguish IBD from IBS or colorectal cancer (CRC).

[0044] Detection of the presence of fecal glucose in stool may indicate issues with sugar absorption in the small intestine, which can lead to osmotic diarrhea and subsequent dehydration.

[0045] LF is an iron-binding glycoprotein that increases in the feces as a result of inflammatory infiltration of granulocytes and neutrophils to the mucosal surface. The presence of LF may be associated with active inflammation.

[0046] Calgranulin C (S100A12) is almost exclusively released by activated granulocytes, and is a useful marker for IBD and the presence of calgranulin-C may may be used to determine if the ostomate has IBD.

[0047] Pyruvate Kinase (M2-PK) is a heterodimer of pyruvate kinase, an enzyme of the glycolytic pathway, and may be expressed in rapidly dividing cells. Originally used as a marker of cell turnover in colorectal cancer, fecal M2-PK may be a potential biomarker of active IBD because of the rapid cell turnover and division seen in IBD. Elevated levels of M2-PK may indicate the presence of IBD and colorectal cancer and, further, may differentiate active inflammation amongst patients with IBD versus those with inactive disease.

[0048] MMPs are also released from the neutrophils of the intestinal mucosa in patients with active IBD. MMP-9 may be used as a fecal biomarker for active ulcerative colitis (UC) and further may be used to distinguish UC from IBS, CD and healthy controls.

[0049] Inflammatory cytokines and chemokines such as Interleukin (IL)- 1, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha, while potentially attractive as biomarkers because of their rolein initiating and maintaining inflammation, are quite unstable and therefore, have a limited role as fecal biomarkers in stools. However, these components may be useful as biomarkers in the dejecta because there may be a reduced period of absorption or breakdown of these components due to the decreased residence time in the body of the ostomate in comparison to formed stool in a non- ostomate.

[0050] Myeloperoxidase (MPO) is an enzyme that is released by neutrophils and is often overexpressed in numerous inflammatory diseases, including IBD. As a result, MPO may serve as a viable, noninvasive fecal biomarker for assessing IBD status. Increased MPO levels have shown correlation with endoscopically-determined inflammation, demonstrating differences in MPO levels with active versus inactive UC and CD. Differences in MPO levels may also distinguish moderate and severe versus endoscopic remission. MPO may also decrease in response to treatment and changes in levels of MPO may indicate effectiveness of such treatment.

[0051] Fecal clearance of plasma alpha 1 -antitrypsin may be used as a measure of protein leakage into the intestinal tract of al -antitrypsin and may be correlated with clinical disease activity in CD and relapse thereof.

[0052] Fecal NGAL is a protein in stool that can indicate inflammation in the gut. NGAL is an antimicrobial protein that is expressed in immune cells, including neutrophils, macrophages, and epithelial cells and may deprive bacteria of iron and thereby limit bacterial growth. NGAL may be unregulated in inflamed colonic epithelium and elevated levels may indicate active IBD, such as UC and CD. NGAL levels may also be elevated in patients with infectious enterocolitis and may be used to distinguish patients with active IBD from healthy controls.

[0053] Fecal hemoglobin is a measurement of blood in the stool that can indicate bleeding in the digestive tract and may be used to screen for colorectal cancer and diagnose other gastrointestinal issues. Detection methods include the gFOBT (guaiac based fecal occult blood test, a peroxidase reaction), immunochemical fecal occult blood test using globin-specific antibodies or ELISA, and bacterial -based biosensors based on engineered Escherichia coli (E.coli). Dejecta hemoglobin may serve as a screen for recurrent CRC, inflammation, and surgical complications such as, for example, bleeding.

[0054] Disclosed herein is a biomarker test device such as a lateral flow immunoassay (LFIA), lateral flow assay (LFA), and the like that may be used to detect an analyte, analytes of interest including but not limited to those described above, glucose, and other materials within dejecta. Such biomarker test devices may be used as a point of care (POC) method to detect molecules in dejecta, incorporated into ostomy pouches or other ostomy system components, or sold as an at- home test to measure the presence of biomarkers (i.e., biomarker molecules) or other components in dejecta and / or changes in levels of such biomarkers. Further, the biomarker test devices may also be used to detect various biomarkers and other molecules in dejecta to better characterize the general composition of the dejecta.

[0055] Referring to FIG. 1, a one piece ostomy system 100 may include an ostomy pouch 102 and a barrier appliance 104. The ostomy pouch 102 may have a first side 106 and a second side 108. When the one-piece ostomy system 100 is used by an ostomate, the first side 106 may face away from a body of the ostomate and the second side 108 may face toward the body of the ostomate. The first side 106 and the second side 108 may be joined (e.g., by fusing, a heat seal, and the like) to one another, for example, along respective perimeters thereof to form a sealed edge 110 of the ostomy pouch 102 that defines a cavity between the first side 106 and the second side 108 for collection of dejecta.

[0056] In other embodiments, the sealed edge 110 spans side and top portions of the perimeters of the first side 106 and the second side 108 but such perimeters are not joined or sealed along a bottom or tail portion 112 of the ostomy pouch 102. In such embodiments, such unsealed portion may include an opening 114. A closing mechanism 116 (e.g., a clamp, a hook and loop closure, etc.) may be disposed on such bottom portion 112 to allow the ostomate to selectively open and close the opening 114. When the opening 114 is closed, the first side 106 and the second side 108 form the cavity into which dejecta may be collected. The opening 114 may be periodically opened so that any dejecta collected in the ostomy pouch 102 may be evacuated through the opening 114 and discarded. The opening 114 may be closed again thereafter.

[0057] The barrier appliance 104 of the one-piece ostomy system 100 may be irremovably secured to the second side 108. The second side 108 and the barrier appliance 104 may include first and second apertures 120, 122, respectively, that may be aligned with one another. In use, a user may enlarge the second aperture 122 according to user’s stoma size by cutting the barrierappliance 104 along one of the cutting guidelines provided on the barrier appliance 104. The first and second apertures 120, 122 may be aligned with a stoma of the ostomate and an adhesive portion on a body face side of the barrier appliance 104 may be used to secure the barrier appliance 104 and, thus, the ostomy pouch 102 to the body of the ostomate so that dejecta discharged from the stoma enters the cavity of the ostomy pouch 102 for collection.

[0058] Referring to FIG. 1A, a two-piece ostomy system 100a may be substantially identical to the one-piece ostomy system 100 except a pouch-side connector flange 124 may be disposed on the second side 108a of the ostomy pouch 102a such that the pouch-side connector flange 124 surrounds the first aperture 120a. A barrier-side connector flange 126 may be disposed on the barrier appliance 104a to surrounds the second aperture 122a. In use, the ostomate may align the second aperture 122a with the stoma and secure an adhesive portion of the barrier appliance 104a to the body. Thereafter, the pouch-side connector flange 124 and the barrier-side connector flange 126 may be joined to one another to secure the ostomy pouch 102a to the barrier appliance 104a and thereby to the body. The pouch-side connector flange 124 and the barrier-side connector flange 126 may be separated from one another to release the ostomy pouch 102a from the barrier appliance 104a in order to, for example, drain or evacuate dejecta collected in the cavity of the ostomy pouch 102a, to replace the ostomy pouch 102a, and the like. In some embodiments, the pouch-side connector flange 124 and barrier-side connector flange 126 may be replaced by releasable adhesives that allow the ostomy pouch 102a to be selectively secured to and released from the barrier appliance 104a.

[0059] Referring to FIGS. 2 and 2A, in one embodiment, a biomarker test device 200 that includes a backing layer 202, a sample pad 204, a conjugate release pad 206, a nitrocellulose membrane 208, and an absorbent pad 210. One or more test lines 212 and a control line 214 are formed in the nitrocellulose membrane 208. The backing layer 202 may be, for example, a paper based material, a plastic based material, and the like. In some embodiments, the nitrocellulose membrane 208 may extend substantially from the sample pad 204 to the absorbent pad 210.

[0060] In some embodiments, the biomarker test device 200 may be a lateral flow immunoassay (LFIA) configured to detect an analyte or analytes of interest including but not limited to those described above regarding dejecta. The conjugate release pad 206 of the LFIA 200 may include at least an antibody conjugated tag comprising, for example, gold, latex, achromophore, a fluorophore, and the like. In some embodiments, one or more such tags may comprise, for example, a nanoparticle.

[0061] During use, a dejecta sample (“sample”), which may contain the analyte of interest is deposited on the sample pad 204 and capillary action draws the sample through the sample pad 204, the conjugate release pad 206, through the nitrocellulose membrane 208, and toward the absorbent pad 210. When the sample is drawn through the conjugate release pad 206, the tag disposed in the conjugate release pad 206 binds the analytes of interest in the sample to form tagbound analytes that flow with the sample toward the absorbent pad 210. The tag-bound analytes and unbound tags flow with the sample as the sample flows toward the absorbent pad.

[0062] In some embodiments, the LFIA 200 may use sandwich binding in which antibodies selected to bind the analytes of interest are disposed in each of the one or more test lines 212. In such embodiments, the tag in the conjugate release pad 206 binds to one end of the analyte (i.e., biomarker) in the sample while the antibody in the test line 212 binds to the other end of the analyte. The combination of such binding and the tag causes the test line 212 to change color to indicate presence of the analytes of interest in the sample.

[0063] In other embodiments, the LFIA 200 may use competitive binding in which additional analytes other than those in the sample are disposed in the test line 212 and only an unbound tag in the sample will be able to bind to such additional analytes and such binding causes the change in the color of the test line 212.

[0064] The tags and the antibodies selected in the LFIA 200 may in accordance with the analyte of interest and may be, for example, Anti-CRP-Mouse-IgG, sheep anti-FITC, and others that would be apparent to one who has ordinary skill in the art.

[0065] Tags (bound and / or unbound) that flow with the sample that reach the control line 214 bind to the control line 214, which changes color to indicate that the tags flowed with the sample to at least the control line 214 and the LFIA 200 has been used correctly.

[0066] In some embodiments, the LFIA 200 may be a positive / negative test model and include a single test line 212. When the positive / negative test model LFIA 200 is used with a sample, the test line 212 changes color to indicate that at least an appropriate concentration of the analyte or biomarker is present in the sample.

[0067] In other embodiments, the LFIA 200 may be semi -quantitative model having a ‘ladder bar’ assay comprising plurality of spaced apart test lines 212. The number of such successive test lines 212 that change color indicates a proportional concentration of the analyte in the sample.

[0068] In another embodiment, the biomarker test device 200 may be an enzyme-based barcode-style lateral-flow assay (LFA) for detection and quantification of glucose at varying concentrations. Such LFA 200 is similar to the LFIA discussed above except the conjugate release pad 206 has glucose oxidase therein instead of tags. In such cases, glucose in the sample deposited on the sample pad 204 and the glucose oxidase from the conjugate release pad 206 flow with the sample and glucose in the sample undergoes a catalytic reaction in the presence of glucose oxidase and water that creates hydrogen peroxide. Such hydrogen peroxide reacts with the one or more of the test lines 212 of the LFA to cause a change in color thereof. The color of the test line 212 changes only if there is enough glucose present in the sample to create sufficient hydrogen peroxide that reaches the test line 212 within a predetermined period of time. In some embodiments, the conjugate release pad 206 may comprise or be replaced with an enzyme pad having the glucose oxidase and a 3, 3, 5, 5, -tetramethylbenzidine (TMB) release pad and the one or more test lines 212 may comprise horseradish peroxidase (HRP). In such embodiments, the hydrogen peroxide produced as described above and the TMB flow toward the one or more test lines 212 and the hydrogen peroxide reacts with the HRP and its colorimetric substrate 3, 3,5,5, - tetramethylbenzidine (TMB) to convert to a blue color complex. The concentration of hydrogen peroxide may determine how many of the one or more test lines 212 have the blue color complex produced thereon. If there is not sufficient glucose in the sample, sufficient hydrogen peroxide may may not be created after the glucose flows past the test line 212. In some embodiments, the predetermined period of time for the hydrogen peroxide to reach a first one of the one or more test lines 212 is approximately nine minutes.

[0069] The enzyme-based barcode-style LFA 200 to detect glucose may also be configured to be semi -quantitative. In such cases, a portion of the hydrogen peroxide is consumed at each test line 212, and successive test lines 212 change color only if there is sufficient analyte concentration in the sample to produce hydrogen peroxide that reaches successive test lines 212. Thus, in some embodiments, the amount of glucose in the sample correlates with the amount of hydrogen-peroxide produced, and the amount of hydrogen-peroxide correlates with the reaction rate of the conversion of TMB by the horseradish peroxidase.

[0070] It should be apparent, the LFA 200 that can assess the level of glucose in a sample of dejecta may be particularly beneficial because the levels of glucose in dejecta may be especially dynamic. That is such levels may change in accordance with the ostomate’s diet, exercise, frequency of eating, and the like. Further, levels of glucose may be an indicator of risk for osmotic diarrhea, as in high output ostomy.

[0071] In addition to detecting glucose as described above, the enzyme-based barcode-style LFA 200 may be adapted to detect the presence or levels of other any hydrogen peroxide producing analyte including, for example, alcohol with alcohol oxidase in the conjugate release pad 206, phosphate with pyruvate oxidase in the conjugate release pad 206, and creatinine with the mixture of creatininase, creatinase, and sarcosine oxidase in the conjugate release pad 206. Adapting the LFA 200 to detect such other compounds may require switching out the conjugate release pad 206 having glucose oxidase with one have a different oxidase selected in accordance with the target analyte.

[0072] In yet another embodiment, the biomarker detection device may be an LFIA 200 configured to detect the presence of lactoferrin. In such embodiments, FITC-labelled antibodies, gold tagged antibodies, and a biotinylated antibody may be deposited in the conjugate release pad 206. Biotin capture molecules are disposed in the one or more test lines 212. When the sample having lactoferrin flows through the conjugate release pad 206, the FITC-labelled antibody binds to first and second ends of the lactoferrin. The gold tagged antibody binds to the FITC labelled antibody bound at the first end of the lactoferrin and the biotinylated antibody binds to the FITC labelled antibody bound at second end of the lactoferrin. When lactoferrin bound in this manner reaches the one or more test lines 212, the biotinylated antibody binds to the biotin capture molecule therein and accumulation of sufficient bound lactoferrin at the one or more test lines 212 cause such test line 212 to change color. In some embodiments, anti-sheep IgG is disposed in the control line 214 that binds with gold-bound sheep anti-FITC this is not bound to lactoferrin and accumulation of such gold-bound sheep anti-FITC at the control line 214 causes a change in color that indicates the sample has reached the control line 214.

[0073] A model dejecta or synthetic feces or fecal sludge may be used to optimize the types and / or amounts of nanoparticles, tags, reactants, and / or enzymes to include in the conjugate release pad 206 and / or the test lines 212, as well as calibrate the biomarker detection device 200 to known concentrations of analytes. In one embodiment, such model dejecta may have a viscosity of between approximately 3,500 and approximately 5,500 centipoise (cps) at 50 rpm and include between approximately 70 and 90 percent water and the remainder being carbohydrates, bacterial biomass, fat, proteins, and inorganics. Table one below shows non-aqueous components of one embodiment of such model dejecta. In some embodiments, the term “buffer” in table one may refer to a solution of water, NaCl, CaC12, and KC1.Table One

[0074] In some embodiments, the pH of the model dejecta may be adjusted to control the rate at which such dejecta flows through the biomarker test device 200. Such rate of flow may be reflected in the intensity over time of color shown at the test line 212 relative to the color of the portions of the biomarker test device 200 between the conjugate release pad 206 and the absorbent pad 210 that are free of the one or more test line 212 and the control line 214. Referring to FIG. 3, the curve 250 shows the intensity of the color shown at the test line 212 over time when the samplecomprised model dejecta having a pH of 6 (i.e., acidic) and the curve 252 shows an intensity of the color test line 212 reached overtime when the sample comprised model dejecta having a pH of 7 (i.e., neutral). Thus, controlling the acidity of the model dejecta may reduce the amount of time required to conduct experiments using such model dejecta.

[0075] In some embodiments, a computational model (e.g., a statistical model, a neural network, and the like) may be trained, configured, or developed to predict a quantity or level of a particular metabolite, protein, or other object in dejecta in accordance with a location of a stoma on an ostomates body from which such dejecta was released. In particular, the computational model may be developed using an input data comprising locations of a plurality of stomas and presence and / or levels of components of the dejecta produced by such plurality of stomas as determined using the biomarker test device 200 as outputs or ground truths. In addition, the training set may include, in some embodiments, additional inputs including environmental factors, genetic factors, and the like. The computational model developed in this manner may be used to provide a prediction or baseline of a metabolite, protein, or other component of dejecta that may be expected based on a stoma location of an individual ostomate.

[0076] In other embodiments, a computation model may be configured to analyze large datasets of dejecta composition as measured by one or more biomarker test devices 200 to identify statistically significant variation in certain metabolites and identify possible correlations with environmental or genetic factors. Applications of this model might be to help identify metabolites for sensing in an ostomy bag, or to improve baselines in LFIA, LFA, ELISA tests, or similar assays and diagnostic protocols.

[0077] In some embodiments, the biomarker test device 200 may be integrated into the ostomy system 100, for example, by attaching at least a portion of the biomarker test device 200 on an interior wall of the ostomy pouch 102, suspending at least a portion of the biomarker test device 200 from an interior wall of the ostomy pouch 102 or within the cavity of the ostomy pouch, or integrated with the barrier appliance 104. In some embodiments, the ostomy pouch 102 may include a clear window and the biomarker test device 200 may be disposed in the ostomy pouch 102 so that at least the one or more test lines 212 of the biomarker test device 200 are visible from an exterior of the ostomy pouch 102 through the clear window.

[0078] In some embodiments, the change in color of the one or more test lines 212 in response to the presence or concentration of an analyte may be detected using an image capture device (e.g., a camera of a mobile phone, a tablet computer, a desktop computer, etc.), for example, if the presence or concentration is too small to cause a color change visually perceptible by a person, to detect small variations in intensity of such color changes, if the wavelength(s) of light reflected or emitted by the test line 212 is / are beyond those perceptible by a human eye, and the like.

[0079] In some embodiments, the biomarker test device 200 may be integrated into a point-of- care or at-home test kit. In some embodiments, the ostomate may see the changes in color of the one or more test lines 212 to assess the presence or concentration of analyte in a sample. In other embodiments, the ostomate may obtain an image of the one or more test lines 212 and send such image to a facility (e.g., a test lab, a medical facility, and the like) and in response receive an analysis of the analyte in the sample.

[0080] Further, in some embodiments, the one or more test lines 212 of the biomarker test device 200 may have electronic polymers disposed therein instead of or in addition to a color changing material. In such embodiments, the amount of analyte in the samples may cause a change in an electrical property such as, for example, conductance, resistance, capacitance, and the like of the test line 212 and such change in the electrical property may be measured to determine presence or a concentration of the analyte. Further, in some embodiments, such one or more test lines 212 may be coupled to an electrical circuit that may generate a signal that may be detected using a measurement device (e.g., a mobile phone, a tablet computer, a desktop computer, etc.) separatee from the biomarker test device 200, wherein the signal indicates the presence or concentration of the analyte. In some embodiments, the circuit may be disposed on the biomarker test device 200. Further, in some embodiments, the measurement device may sense the signal wirelessly.

[0081] From the foregoing it will be observed that numerous modifications and variations can be effectuated without departing from the true spirit and scope of the novel and non-obvious aspects of the present disclosure. It is to be understood that no limitation with respect to the specific embodiments illustrated is intended or should be inferred. The disclosure is intended to cover by the appended claims all such modifications as fall within the scope of the claims.

[0082] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0083] The use of the terms “a” and “an” and “the” and similar references in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the disclosure and does not pose a limitation on the scope of the disclosure unless otherwise claimed. No language in the specification should be construed as indicating any nonclaimed element as essential to the practice of the disclosure.

[0084] Numerous modifications to the present disclosure will be apparent to those skilled in the art in view of the foregoing description. It should be understood that the illustrated embodiments are exemplary only, and should not be taken as limiting the scope of the disclosure.

Claims

WE CLAIM:

1. A device adapted to detect a presence and / or concentration of a nutrient and / or biomarker in a stoma dejecta sample, comprising: a backing layer; a sample pad disposed on the backing layer; a conjugate release pad in fluid communication with the sample pad; a first material disposed in the conjugate release pad, wherein the first material is selected in accordance with the nutrient or biomarker; and a test line in fluid communication with the conjugate release pad; wherein the device is configured to cause the stoma dejecta sample deposited on the sample pad to flow through the conjugate release pad so that the nutrient or biomarker interacts with the first material to form a second material that flows to the test line, wherein the test line is configured to change color in response to accumulation of the second material thereon.

2. The device of claim 1, wherein the device is configured to detect presence or concentration of glucose, alcohol, pyruvate oxidase, or creatinine.

3. The device of claim 2, wherein the first material includes an enzyme.

4. The device of any one of claim 1-3, wherein the second material includes hydrogen peroxide.

5. The device of claim 1, wherein the biomarker includes a protein.

6. The device of claim 5, wherein the first material includes a nanoparticle.

7. The device of claim 1, wherein the biomarker includes one or more of polymorphonuclear neutrophil elastase, lactoferrin, macrophage cytosolic protein S100A12 (calgranulin C), pyruvate kinase, interleukin 1, interleukin 6, interleukin 8, tumor necrosis factor a, glucose, myeloperoxidase, al -antitrypsin, neutrophil gelatinase-associated lipocalin, hemoglobin, or a matrix metalloprotease.

8. The device of claim 1, wherein the biomarker includes one of inflammatory cytokines and chemokines.

9. The device of any one of claims 1-8, wherein the device is a Lateral Flow Immunoassay (LFIA).

10. The device of any one of claims 1-8, wherein the device is an Enzyme-based Barcode-style Lateral-flow Assay.

11. A method of detecting a presence and / or concentration of a nutrient and / or biomarker in a stoma dejecta sample using a biomarker testing device having a sample pad, a conjugate release pad having a first material disposed therein, and a test line, comprising: depositing the stoma dejecta on sample the sample pad; causing the stoma dejecta to flow through the conjugate release pad; causing an interaction of the nutrient or biomarker interacts with the first material to form a second material, wherein the first material is selected in accordance with the nutrient or biomarker; causing the second material to accumulate at the test line, and thereby causing the test line to change color.

12. The method of claim 11, wherein the nutrient or biomarker includes glucose, alcohol, pyruvate oxidase, or creatinine.

13. The method of claim 12, wherein the first material includes an enzyme.

14. The method of any one of claims 11-13, wherein the second material includes hydrogen peroxide.

15. The method of claim 11, wherein the biomarker includes a protein.

16. The method of claim 14, wherein the first material includes a nanoparticle.

17. The method of claim 11, wherein the biomarker includes one or more of polymorphonuclear neutrophil elastase, lactoferrin, macrophage cytosolic protein S100A12 (calgranulin C), pyruvate kinase, interleukin 1, interleukin 6, interleukin 8, tumor necrosis factor a, glucose, myeloperoxidase, al -antitrypsin, neutrophil gelatinase-associated lipocalin, hemoglobin, or a matrix metalloprotease.

18. The method of claim 11, wherein the biomarker includes one of inflammatory cytokines and chemokines.

19. A model dejecta composition for optimizing and calibrating the device of any one of claims 1-10, comprising: between 70 and 90 percent water; the nutrient and / or biomarker; and one or more of carbohydrates, bacterial biomass, fat, proteins, and inorganics, wherein the model dejecta composition has a viscosity of between approximately 3,500 and approximately 5,500 cPs at 50 rpm.

20. The model dejecta composition of claim 19, wherein the model dejecta composition is acidic.

21. The model dejecta composition of claim 19 or claim 20, wherein the model dejecta composition comprises 6.8g NaCl, 6.8g KC1, 3.4g CaC12 per liter of water.

22. The model dejecta composition of any one of claims 19-21, wherein the model dejecta composition comprises 333g rice flour per liter of water.

23. The model dejecta composition of any one of claims 19-22, wherein the model dejecta composition comprises 100g polyethylene glycerol per liter of water.

24. The model dejecta composition of any one of claims 19-23, wherein the model dejecta composition comprises 5g collagen per liter of water or 10% cellulose per liter of water.

25. The model dejecta of any one of claims 19-24, wherein the model dejecta composition comprises 60 grams peanut oil per liter of water.

26. The model dejecta of any one of claims 19-25, wherein the model dejecta composition comprises 120g yeast per liter of water.

27. The model dejcta of any one of claims 19-26, wherein the model dejecta composition comprises a biomarker or a nutrient.

28. An ostomy system comprising an ostomy pouch and a barrier appliance, wherein the device of any one of claims 1-10 is incorporated into the ostomy system.

29. The ostomy system of claim 28, wherein the device is disposed in the ostomy pouch.30 The ostomy system of clam 28 or 29, wherein the ostomy pouch includes a clear window and the device is viewable through the clear window from an exterior of the ostomy pouch.

31. An at-home test kit comprising the device of any one of claims 1-10.

32. A computational model to predict a presence and / or concentration of a nutrient and / or biomarker in dejecta in accordance with a placement of a stoma on ostomate, wherein the computation model is configured using a plurality of the devices of any one of claims 1-10 toanalyze a component of dejecta associated with a corresponding plurality of stomas disposed in different locations.

33. A computational model to determine a correlation between a presence and / or concentration of a nutrient and / or a biomarker and environmental and / or genetic factors, wherein the computational model is configured using a plurality of devices of any one of claim 1-10 to analyze a component of dejecta from a plurality of ostomates associated with a plurality of environmental and / or genetic factors.

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