Systems and methods for biomarker measurement using smartphones and the like

Abstract

The disclosure relates to systems and methods for biomarker measurement using smartphones. The systems include smartphones having application software configured to obtain images of biomarker test kits and deliver those images to back-end systems to analyze image data obtained from the smartphones.

Description

SYSTEMS AND METHODS FOR BIOMARKER MEASUREMENT USING SMARTPHONES AND THE LIKEFIELD

[0001] The disclosure relates to systems and methods for biomarker measurement using smartphones. The systems include smartphones having application software configured to obtain images of biomarker test kits and deliver those images to back-end systems to analyze image data obtained from the smartphones.BACKGROUND

[0002] Traditional diagnostic testing methods require individuals to visit doctors’ offices in order to obtain requisitions and then subsequently attend a testing laboratory to undertake the diagnostic test. For many tests, this puts unnecessary costs on health care systems as many hands become involved in what can be considered as highly routine and simple tests. Such methods can also be highly inconvenient to patients who often must travel between locations to complete the tests. After the tests are complete, there are additional delays in processing the results and ultimately delivering the results to various stakeholders including the patient, care-professionals and others.

[0003] Moreover, access to many diagnostic tests is limited, particularly in remote or underserved areas. Further, patients often have fear and / or anxiety in attending facilities to have tests completed.

[0004] Accordingly, there has been a need for improved diagnostic test systems that improve the efficiencies of completing tests and returning data to stakeholders.SUMMARY

[0005] In accordance with the disclosure, there is provided a system for obtaining and analyzing images from a biomarker test comprising: a smartphone having a camera and a smartphone processor configured with smartphone application software and having non- transitory memory including instructions configured to: prompt a user to initiate image acquisition from the biomarker test; activate the camera; prompt the user to position the biomarker test on a surface and position the camera over the biomarker test; obtain at leastone captured image; and, deliver the at least one captured image to a back-end computer system over the internet; wherein the back-end computer system includes a back-end processor configured with back-end application software having non-transitory memory including instructions configured to: receive the at least one captured images; conduct a color-density analysis of the captured images; and, compare the color-density analysis obtained against a database of color-density results and obtain a diagnostic test result.

[0006] In various embodiments:• the back-end processor is configured with instructions to report the diagnostic test result to one or more stakeholders over the internet.• the smartphone processor is configured with instructions to obtain first and second captured images in succession.• the first and second captured images are RAW data images.• the smartphone processor is configured with instructions to activate a camera flash during acquisition of the first or second captured image.• the smartphone processor is configured with instructions to verify a brightness variation between the first and second captured images.• the smartphone processor is configured with instructions to provide an error signal if the brightness variation between the first and second captured images is greater than 3.4.• the back-end processor is configured with instructions to select a green channel from the first and second images, convert to greyscale and apply a region of interest.• the back-end computer system includes a back-end database storing biomarker test parameters and the back-end processor is configured with instructions to search the back-end database to access a testing protocol specific to a biomarker test kit. the back-end computer system is configured with instructions to analyze a relative color-density and correlate the color-density to a quantitative level of analyte from a biomarker test.

[0007] In another aspect, the disclosure provides a method of obtaining and analyzing images from a biomarker test comprising the steps of: from a smartphone having a camera and a smartphone processor configured with application software and having non-transitory memory including instructions: prompting a user to initiate image acquisition; activating the smartphone camera; prompting the user to position a biomarker test on a surface and position the camera over the completed test; obtaining at least one capture image; and, delivering captured images to a back-end system over the internet.

[0008] In various embodiments, the method includes:• reporting the diagnostic test result to one or more stakeholders over the internet.• obtaining first and second captured images in succession.• activating a camera flash during acquisition of the first or second captured image.• verifying a brightness variation between the first and second captured images.• providing an error signal if the brightness variation between the first and second captured images is greater than 3.4.• selecting a green channel from the first and second images, convert to greyscale and apply a region of interest.• searching a back-end database to access a testing protocol specific to a biomarker test kit.• analyzing a relative color-density and correlating the color-density to a quantitative level of analyte from a biomarker test.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Various objects, features and advantages of the disclosure will be apparent from the following description of particular embodiments, as illustrated in the accompanying drawings. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of various embodiments of the disclosure. Similar reference numerals indicate similar components.Figure 1 is a schematic plan view of a typical linear flow assay (LFA) test device in accordance with the prior art showing the visual results of diagnostic test including a positive (A), a negative (B) and an inconclusive (C) test.Figure 2 is a schematic side view of a typical mechanism of operation of an LFA in accordance with the prior art showing the configuration of a test kit before use (A) and the binding and migration of analytes across a test kit during a test (B).Figure 3 is a schematic side view of a smartphone aligned to image a LFA test device in accordance with the disclosure.Figure 4 is a diagram showing a communication system to obtain, analyze and report test results to stakeholders in accordance with the disclosure.Figure 5 is a flowchart showing steps of obtaining, analyzing and reporting test results to stakeholders in accordance with the disclosure.Figure 6 is a representative example of a color-density vs. distance profile for T and C lines in accordance with the disclosure.Figure 7 is a flowchart showing steps of identifying a bioassay test kit and activating a test protocol based on known or generic test protocols in accordance with the disclosure.DETAILED DESCRIPTION

[0010] With reference to the figures, systems and methods for conducting biomarker tests are described.Terminology

[0011] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the disclosure. As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be understood that the terms “comprises” and / or “comprising,” when used in this specification, specify the presence of stated features, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or moreother features, steps, operations, elements, components, and / or groups thereof. As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items.

[0012] Spatially relative terms may be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a feature in the figures is inverted, elements described as “under” or “beneath” other elements or features would then be oriented “over” the other elements or features. Thus, the exemplary term “under” can encompass both an orientation of over and under. A feature may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly. Similarly, the terms “upwardly”, “downwardly”, “vertical”, “horizontal” and the like are used herein for the purpose of explanation only unless specifically indicated otherwise.

[0013] It will be understood that when an element is referred to as being “on”, “attached” to, “connected” to, “coupled” with, “contacting”, etc., another element, it can be directly on, attached to, connected to, coupled with or contacting the other element or intervening elements may also be present. In contrast, when an element is referred to as being, for example, “directly on”, “directly attached” to, “directly connected” to, “directly coupled” with or “directly contacting” another element, there are no intervening elements present.

[0014] It will be understood that, although the terms “first”, “second”, etc. may be used herein to describe various elements, components, etc., these elements, components, etc. should not be limited by these terms. These terms are only used to distinguish one element, component, etc. from another element, component. Thus, a “first” element, or component discussed herein could also be termed a “second” element or component without departing from the teachings of the present disclosure. In addition, the sequence of operations (or steps) is not limited to the order presented in the claims or figures unless specifically indicated otherwise.

[0015] Other than described herein, or unless otherwise expressly specified, all of the numerical ranges, amounts, values and percentages, such as those for amounts of materials, elemental contents, times and temperatures, ratios of amounts, and others, in the following portion of the specification and attached claims may be read as if prefaced by the word“about” even though the term “about” may not expressly appear with the value, amount, or range. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0016] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0017] Various aspects of the disclosure will now be described with reference to the figures. The disclosure may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art. Moreover, the drawings are not necessarily drawn to scale and are intended to emphasize principles of operation rather than precise dimensions.Introduction

[0018] Test kits for assaying various biomarkers are known. For example, during the Covid- 19 pandemic, the world became particularly familiar with conducting lateral flow assay (LFA) tests.

[0019] As is known, an LFA test is typically conducted by accessing a test kit, obtaining a biological sample, introducing the biological sample potentially containing an analyte of interest to a lateral flow medium, introducing buffer and waiting a period of time for the biological sample to react with a test line and a control line to produce a test result.

[0020] By way of example, as shown in Figures 1 and 2, a test kit 10 generally includes a sample pad 10b, a conjugate release pad 10c, a membrane 10d, a test line 10e, a control line 10f and an absorbent pad 10g mounted to a backing card 10a for stability and handling.

[0021] As shown in Figure 2A, a liquid sample containing the analyte of interest Z is applied to the sample pad 10a which can then diffuse / move via capillary action through the various zones of the membrane where the analyte can interact with various capture reagents contained in different zones of the LFA.

[0022] For example, the test kit may be configured with antibodies Y that are specific to the analyte Z which bind the analyte as the sample migrates across the strip. The antibodies Y are typically conjugated to colored or fluorescent particles-most commonly colloidal gold and latex microspheres (marker particles) O shown as a Y-0 complex within the conjugate release pad 10c. As the analyte Z migrates across the membrane 10d, it binds to the Y-0 complex to form a Y-O-Z complex which migrates to the test line 10e. The test line is conjugated with an anti-analyte antibody Y1 which can form a Y1-Z-Y-0 complex. Binding at the test line results in a visible line.

[0023] The control line 10f is conjugated with antibodies Y2 specific to Y such that mobilized Y-0 complexes will bind to the control line as Y2-Y-0 complexes. Binding at the control line results in a visible line.

[0024] If the analyte of interest is present in the sample, the Y-Z-0 complex forms which can then be captured at the test line as a Y1-Y-Z-0 complex. Y-0 will also be bound at the control line as Y2-Y-O. The combination of these two visible lines indicates a positive test (Figures 1A and 2B).

[0025] If the analyte is not present in the sample, the Y-Z-0 complex does not form at the test line while Y-0 binding occurs at the control line as Y2-Y-0 which indicates a negative test (Figure 1 B).

[0026] Weakly appearing lines can indicate an inconclusive test (Figure 1C).

[0027] Figures 1 and 2 show a simple positive / negative test system. In other test kits, the relative intensity of lines and / or the position of lines can be an indication of the relative concentration of an analyte.

[0028] Further, other test kits can enable multiple analytes to be measured simultaneously under the same conditions, by providing additional test lines of antibodies specific to different analytes immobilized in an array format.

[0029] Further, in some tests, multiple test lines loaded with the same antibody can be used for semi-quantitative assays. In this case, a ‘ladder bars’ assay is based on the stepwise capture of colorimetric conjugate-antigen complexes by the immobilized antibody on each successive line, where the number of lines appearing on the strip is directly proportional to the concentration of the analyte.

[0030] Test results can be interpreted by eye or by using dedicated readers. While test results can often be readily understood, there can be an element of subjectivity particularly in the case of interpretation by eye. Furthermore, dedicated readers may not be readily available and / or may be prohibitively expensive. Moreover, the reporting of test results to stakeholders can be non-existent and / or subject to significant delays.

[0031] In accordance with this description, the inventors have recognized that smartphones can be utilized to address these challenges which can improve the efficiency and accuracy of conducting tests, interpreting results and delivering those results to stakeholders.

[0032] As shown in Figures 3 and 4, in one embodiment, a smartphone 30 having a camera application software configured to obtain images of test results from a test kit 10, together with back-end application can be used to collect and analyze test kit data and deliver that analyzed data to stakeholders.

[0033] The system is described via a representative example, where a patient and / or their caregiver (a “user”) wishes to complete a test and have those results delivered back to the user and / or to a medical team.

[0034] A test is completed in the usual manner and a result is obtained where the test line and control line are both visible (e.g. a positive test).

[0035] The user accesses a smart phone 30 having application software configured to obtain images of the test kit and automatically deliver those images to a back-end system having back-end software to receive and interpret image results.

[0036] In one embodiment, the smart phone application instructs the user to place the completed test on a surface in a manner that enables one or more images of the test kit to be obtained to enable accurate interpretation of image(s) of the test kit.

[0037] In a typical system, the user will be prompted through a series of instructions to position the smartphone camera over the completed test wherein one or more images of the test kit will be obtained. The images may be automatically delivered via the internet to a back- end system 40n where the images are interpreted and a result obtained. Such results may be automatically reported back to any one of or a combination of the patient 40a, a caregiver 40b, a health care provider 40c and / or a health authority 40d as shown in Figure 4.

[0038] In various embodiments as shown in Figure 5, upon activation of the smartphone application the smartphone application is configured to: a. prompt the user to complete a test. b. prompt the user to confirm that the test is completed. c. automatically activate the smartphone camera. d. prompt the user to position the completed test on a surface and position the camera in a particular manner over the completed test. e. automatically obtain images and / or prompt a user to obtain images. f. automatically deliver captured images to a back-end system over the internet.

[0039] In various embodiments, the smartphone application software may be configured to: a. activate a smartphone flash 30a and deliver a known flash intensity as images are being collected. b. prompt the user to locate the test kit at a specific distance and orientation to the camera.

[0040] Importantly, as the location and lighting conditions that may be present after completion of a test will be highly variable, the smartphone is configured to capture at least two RAW images of an LFA test line. In one embodiment, a first image is taken without a flash and a second image is taken with a flash resulting in an image pair. The images of the image pair are taken in quick succession to eliminate movement variation between the images.

[0041] Image pairs, where the brightness variation between the images does not exceed a threshold ratio, for example a ratio of 3.4 of flash image over a no flash image, an image pair may be rejected and the user prompted to retake the images.

[0042] If the images are satisfactory, the selected images are uploaded to a back-end database for analysis.

[0043] In one embodiment, the green channel is selected from each image, converted to grayscale, and a ROI (region of interest) is applied to the LFA test window. A gray intensity profile is then plotted based on the ROI line.

[0044] Figure 6 shows an analysis of an image wherein the color intensity as a function of distance (in pixels) from an origin is calculated. As shown, a maximum peak value is identified for the control (C) and the test (T) lines of the LFA and a ratio of average T / C ratio is used to calculate positive / negative results for those lines.

[0045] In various embodiments, where the test kit is configured to determine analyte concentration, the relative intensity of lines may be correlated to an analyte concentration.

[0046] The positive / negative and / or quantitative measurement is then sent back to the user’s smartphone, and / or to other stakeholders if permissions have been granted.

[0047] In various embodiments, the smartphone application software is configured to automatically recognize the type of test being conducted which may be determined by ID markings / codes on the test kit, such as QR codes (Figure 3).

[0048] By recognizing a specific test kit (as shown in Figure 7), which may be specific to a manufacturer and / or may be a test kit for a specific analyte or multiple analytes, the smartphone application software may deliver prompts to the user specific to that test kit, follow different image capture protocols and / or mark the image data as specific to a type of test, enabling the back-end to then recognize the images and analyze the image(s) according to a preferred analysis protocol for that specific test kit. If an ID code is not recognized, a generic test protocol may be activated that may simply assess basic test results such as standard T and C lines on an LFA.

[0049] Non-limiting examples of analytes that can be analyzed using the system may include Ferritin(iron), Progesterone, Estradiol, HcG (Human Gonadotropin), Luteinizing Hormone (LH), ApoB, Total Cholesterol, LDL, HDL, Haemoglobin, Vitamin D, etc.

[0050] Although the present disclosure has been described and illustrated with respect to preferred embodiments and preferred uses thereof, it is not to be so limited since modifications and changes can be made therein which are within the full, intended scope of the disclosure as understood by those skilled in the art.

Claims

CLAIMS1. A system for obtaining and analyzing images from a biomarker test comprising: a smartphone having a camera and a smartphone processor configured with smartphone application software and having non-transitory memory including instructions configured to:(1) prompt a user to initiate image acquisition from the biomarker test;(2) activate the camera;(3) prompt the user to position the biomarker test on a surface and position the camera over the biomarker test;(4) obtain at least one captured image; and,(5) deliver the at least one captured image to a back-end computer system over the internet; ii) wherein the back-end computer system includes a back-end processor configured with back-end application software having non-transitory memory including instructions configured to:(1) receive the at least one captured images;(2) conduct a color-density analysis of the captured images; and,(3) compare the color-density analysis obtained from ii)(2) against a database of color-density results and obtain a diagnostic test result.

2. The system as in claim 1 where the back-end processor is configured with instructions to report the diagnostic test result from ii)(3) to one or more stakeholders over the internet.

3. The system as in claim 1 where the smartphone processor is configured with instructions to obtain first and second captured images in succession.

4. The system as in claim 3 where the first and second captured images are RAW data images.

5. The system as in claim 3 where the smartphone processor is configured with instructions to activate a camera flash during acquisition of the first or second captured image.

6. The system as in claim 3 where the smartphone processor is configured with instructions to verify a brightness variation between the first and second captured images.

7. The system as in claim 6 where the smartphone processor is configured with instructions to provide an error signal if the brightness variation between the first and second captured images is greater than 3.4.

8. The system as in claim 3 wherein the back-end processor is configured with instructions to select a green channel from the first and second images, convert to greyscale and apply a region of interest.

9. The system as in claim 1 wherein the back-end computer system includes a back- end database storing biomarker test parameters and the back-end processor is configured with instructions to search the back-end database to access a testing protocol specific to a biomarker test kit.

10. The system as in any one of claims 1-9 wherein the back-end computer system is configured with instructions to analyze a relative color-density and correlate the colordensity to a quantitative level of analyte from a biomarker test.

11. A method of obtaining and analyzing images from a biomarker test comprising the steps of:from a smartphone having a camera and a smartphone processor configured with application software and having non-transitory memory including instructions:(1) prompting a user to initiate image acquisition;(2) activating the smartphone camera;(3) prompting the user to position a biomarker test on a surface and position the camera over the completed test;(4) obtaining at least one capture image; and,(5) delivering captured images to a back-end system over the internet.

12. The method as in claim 11 further comprising the step of reporting the diagnostic test result to one or more stakeholders over the internet.

13. The method as in claim 11 or claim 12 further comprising the step of obtaining first and second captured images in succession.

14. The method as in claim 13 further comprising the step of activating a camera flash during acquisition of the first or second captured image.

15. The method as in claim 14 further comprising the step of verifying a brightness variation between the first and second captured images.

16. The method as in claim 15 further comprising the step of providing an error signal if the brightness variation between the first and second captured images is greater than 3.4.

17. The method as in claim 13 further comprising the step of selecting a green channel from the first and second images, convert to greyscale and apply a region of interest.

18. The method as in claim 11 further comprising the step of searching a back-end database to access a testing protocol specific to a biomarker test kit.

19. The method as in claim 11 further comprising the steps of analyzing a relative color-density and correlating the color-density to a quantitative level of analyte from a biomarker test.

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