Pathogen monitoring system and method

Abstract

The portable sensor system (100) includes a containment portion (102) and a sensor portion (104). The containment portion (102) may include a shipping container such as a jar. The sensor portion (104) may include a cap (106) that may be selectively coupled with the containment portion (102). The cap (106) may include a battery module (108), a processing module (110), and a sensor module (112). In a specific example, the portable sensor system (100) may be provided in four layers (a battery module layer (108), a processing module layer (110), a sensor module layer (112), and a coupling mechanism layer (114). The coupling mechanism layer (114) may be configured to selectively lock the cap (106) to the containment portion (102). In a more specific example, one or more of the layers (108, 110, 112, 114) may be 3D printed. The portable sensor system (100) may be utilized as a smart-monitoring device for the presence of a pathogenic bacteria in the shipping container.

Description

Attorney Docket No. 70673-02PATHOGEN MONITORING SYSTEM AND METHODCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 636,908 filed April 22, 2024, the entirety of which is hereby incorporated by reference.GOVERNMENT RIGHTS

[0002] This invention was made with government support under 59-8072-1-002 awarded by the United States Department of Agriculture Agricultural Research Service. The government has certain rights in the invention.FIELD

[0003] The disclosure generally relates to monitoring systems and, more particularly, to pathogen monitoring systems.INTRODUCTION

[0004] This section provides background information related to the present disclosure which is not necessarily prior art.

[0005] The Centers for Disease Control and Prevention estimates that the annual toll of foodborne illness in the United States is significant, with 48 million cases, 128,000 hospitalizations, and 3,000 deaths, resulting in a staggering $152 billion in medical costs, lost productivity, legal expenses, and damage to brand reputation. Detection and identification of pathogens is the first step for mitigating foodborne disease outbreaks in the subsequent flow of farm to fork of food supply chain. There is a universal demand among regulators, food producers, processors, and researchers for swift, accurate, and precise detection methods for foodborne pathogens and other hazards associated with food. The standard practice involves collecting food product samples and sending them to a central laboratory for microbiological testing. However, this process introduces delays in obtaining the microbiological testing results and subsequently affects the timely delivery of food products to consumers. Traditional methods like culturing and plating are considered gold standards due to their ability to detect low numbers of pathogenic cells through selective enrichment, however they have drawbacks, particularly the extended time required for results — often more than fiveAttorney Docket No. 70673-02 days. The predominant tools employed in food microbiology for the detection and identification of pathogens rely on the physiological, immunological, or genetic properties of microorganisms. These methods encompass the examination of substrate utilization's metabolic characteristics, the scrutiny of signature molecules through antibodies, DNA analysis, and the study of pathogen interaction with eukaryotic cells. Subsequent sections of this proposal will delve into advancements within these domains. Nevertheless, there is considerable interest in the adoption of recently introduced label-free and reagent-less biosensors that leverage the biophysical properties of samples. For certain food samples, conventional testing flow for food safety protocol requires sampling the food from the production facility and either conduct the testing on-site or ship them to a third party or government / regulatory laboratories. Many medium to small size production facilities generally don’t operate then' own testing facilities due to operation cost and food sample testing results can be delayed due to shipping of samples to the various external laboratories. Portable sensors and detection systems are becoming popular in food safety areas due to the user- friendly operation, lower cost per test, and integration of internet of things (IOT) to provide broader understanding in terms of food safety concerns in the manufacturing plant. Some examples include utilizing smartphone-based sensors to provide quantitative concentration of pathogens from lateral flow strips and bacterial colonies, portable sensors that can provide ATP-based or bioluminescence phase-based pathogen detection, IOT based temperature monitoring for potential risk assessment in food safety. Known sensor systems rely on end point detection. This loses out on valuable time for detection while shipping is happening. This loss of time may add an additional one to two days in time to detection. In other words, you either measure at a production facility or at a USDA FSIS or other laboratories. Once the sample arrives, they need to go through processes for sample preparations and enrichment.

[0006] Accordingly, there is a continuing need for a portable sensor that may detect the presence of a pathogenic bacteria during the shipping time from a production facility to a testing laboratory.SUMMARY

[0007] In concordance with the present disclosure, a portable sensor that may detect the presence of a pathogenic bacteria during the shipping time from a production facility to a testing laboratory, has surprisingly been discovered.Attorney Docket No. 70673-02

[0008] A portable sensor that can be integrated into conventional shipping container of food sample and turn the container into a smart-monitoring device for the presence of a pathogenic bacteria in the shipping container. The portable sensor system may include a containment portion and a sensor portion. The containment portion may include a shipping container such as ajar. The sensor portion may include a cap that may be selectively coupled with the containment portion. The cap may include a battery, a processing unit, and a sensor unit.

[0009] As a non-limiting example, a microcontroller and a Silicon photomultiplier (SiPM)- based detector was integrated into a stand-alone cap that replaced an original bottle cap of the sample container. The portable sensor system may also include a reporter phage, which may then provide historical data of the optical readout from the sensor throughout the whole shipping period to the third-party lab or government laboratories. Advantageously, since these locations generally receive large quantity of samples to be tested, the portable sensor may provide a quick readout of which samples to be prioritized screened for a confirmatory study and subsequent intervention procedure.

[0010] The portable sensor system may provide real-time and in-transit monitoring of the contamination status of the food sample, specifically targeting E. coli O157:H7, through a bioluminescence assay. The assay may exclusively target a target pathogen and, when detected, produces minimal luminescence. As the sample is transported in the container, the number of bacterial cells multiplies, and once the luminescent signal reaches a predefined threshold, the sensor reports the results, such as via Bluetooth or other means of information transmission.

[0011] Further areas of applicability will become apparent from the description provided herein. The description and specific examples in this summary are intended for purposes of illustration only and are not intended to limit the scope of the present disclosure.DRAWINGS

[0011] The drawings described herein are for illustrative purposes only of selected embodiments and not all possible implementations and arc not intended to limit the scope of the present disclosure.

[0012] FIG. 1 is a flow diagram illustrating the process for using a portable sensor system, otherwise known as MP ACT, further depicting the introduction of a bacteriophase-basedAttorney Docket No. 70673-02 assay to multiply the pathogen, a Lux gene is introduced via phage if target bacteria is present in the food sample, and a light is monitored, according to one embodiment of the present disclosure;

[0013] FIG. 2 is a front perspective view of the portable sensor system, according to one embodiment of the present disclosure;

[0014] FIG. 3 is a functional block diagram of the portable sensor system, according to one embodiment of the present disclosure;

[0015] FIG. 4 is a flowchart of the testing procedure of the portable sensor system, further depicting a dilution procedure followed by a light measurement procedure, according to one embodiment of the present disclosure;

[0016] FIG. 5A is plot diagram illustrating the relative light sensing performance of the portable sensor system compared to a lab luminometer, according to one embodiment of the present disclosure;

[0017] FIG. 5B is a side elevational schematic diagram of the portable sensor system having a lens to effectively make the sensor area larger by focusing more light onto the sensor, further depicting where a diffusive cone provides a chance for suboptimal rays to eventually find the sensor as well, according to one embodiment of the present disclosure;

[0018] FIG. 6 is a front perspective view of the portable sensor system, according to one embodiment of the present disclosure;

[0019] FIG. 7 is an exploded top perspective view of the cap of the portable sensor system, according to one embodiment of the present disclosure;

[0020] FIG. 8 is an exploded front perspective view of the portable sensor system, further depicting the containment portion without an opaque cover, according to one embodiment of the present disclosure;

[0021] FIG. 9 is an exploded front perspective view of the portable sensor system, further depicting the containment portion with the opaque cover to militate against environmental exposure, according to one embodiment of the present disclosure;

[0022] FIG. 10 is a front perspective view of the portable sensor system, according to one embodiment of the present disclosure;

[0023] FIG. 11 is an exploded front perspective view of the cap of the portable sensor system, according to one embodiment of the present disclosure;Attorney Docket No. 70673-02

[0024] FIG. 12 is a circuit diagram of the portable sensor system, according to one embodiment of the present disclosure;

[0025] FIG. 13 is a top perspective view of the processing layer of the cap, according to one embodiment of the present disclosure;

[0026] FIG. 14 is a top perspective view of the sensor layer of the cap, according to one embodiment of the present disclosure;

[0027] FIG. 15 is a front perspective view of the cap of the portable sensor system, according to one embodiment of the present disclosure;

[0028] FIG. 16 is a front perspective view of the cap of the portable sensor system without an outer cover, according to one embodiment of the present disclosure;

[0029] FIG. 17 is a real' elevational view of the cap of the portable sensor system without a cover, according to one embodiment of the present disclosure;

[0030] FIG. 18 is a plot diagram illustrating the sensitivity of the portable sensor system, according to one embodiment of the present disclosure; and

[0031] FIG. 19 is a flowchart depicting a method of using the portable sensor system, according to one embodiment of the present disclosure.DETAILED DESCRIPTION

[0032] The following description of technology is merely exemplary in nature of the subject matter, manufacture and use of one or more inventions, and is not intended to limit the scope, application, or uses of any specific invention claimed in this application or in such other applications as may be filed claiming priority to this application, or patents issuing therefrom. Regarding methods disclosed, the order of the steps presented is exemplary in nature, and thus, the order of the steps can be different in various embodiments, including where certain steps can be simultaneously performed. “A” and “an” as used herein indicate “at least one” of the item is present; a plurality of such items may be present, when possible. Except where otherwise expressly indicated, all numerical quantities in this description arc to be understood as modified by the word “about” and all geometric and spatial descriptors are to be understood as modified by the word “substantially” in describing the broadest scope of the technology. “About” when applied to numerical values indicates that the calculation or the measurement allows some slight imprecision in the value (with some approach to exactness in the value; approximately or reasonably close to theAttorney Docket No. 70673-02 value; nearly). If, for some reason, the imprecision provided by “about” and / or “substantially” is not otherwise understood in the art with this ordinary meaning, then “about” and / or “substantially” as used herein indicates at least variations that may arise from ordinary methods of measuring or using such parameters.

[0033] Although the open-ended term “comprising,” as a synonym of non-restrictive terms such as including, containing, or having, is used herein to describe and claim embodiments of the present technology, embodiments may alternatively be described using more limiting terms such as “consisting of’ or “consisting essentially of.” Thus, for any given embodiment reciting materials, components, or process steps, the present technology also specifically includes embodiments consisting of, or consisting essentially of, such materials, components, or process steps excluding additional materials, components or processes (for consisting of) and excluding additional materials, components or processes affecting the significant properties of the embodiment (for consisting essentially of), even though such additional materials, components or processes are not explicitly recited in this application. For example, recitation of a composition or process reciting elements A, B and C specifically envisions embodiments consisting of, and consisting essentially of, A, B and C, excluding an element D that may be recited in the art, even though element D is not explicitly described as being excluded herein.

[0034] As referred to herein, disclosures of ranges are, unless specified otherwise, inclusive of endpoints and include all distinct values and further divided ranges within the entire range. Thus, for example, a range of “from A to B” or “from about A to about B” is inclusive of A and of B. Disclosure of values and ranges of values for specific parameters (such as amounts, weight percentages, etc.) are not exclusive of other values and ranges of values useful herein. It is envisioned that two or more specific exemplified values for a given parameter may define endpoints for a range of values that may be claimed for the parameter. For example, if Parameter X is exemplified herein to have value A and also exemplified to have value Z, it is envisioned that Parameter X may have a range of values from about A to about Z. Similarly, it is envisioned that disclosure of two or more ranges of values for a parameter (whether such ranges arc nested, overlapping, or distinct) subsume all possible combination of ranges for the value that might be claimed using endpoints of the disclosed ranges. For example, if Parameter X is exemplified herein to have values in the range of 1-10, or 2-9, or 3-8, it is also envisioned that Parameter X may have other ranges of values including 1-9, 1-8, 1-3, 1-2, 2-10, 2-8, 2-3, 3-10, 3-9, and so on.Attorney Docket No. 70673-02

[0035] When an element or layer is referred to as being “on,” “engaged to,” “connected to,” or “coupled to” another element or layer, it may be directly on, engaged, connected, or coupled to the other element or layer, or intervening elements or layers may be present. In contrast, when an element is referred to as being “directly on,” “directly engaged to,” “directly connected to” or “directly coupled to” another element or layer, there may be no intervening elements or layers present. Other words used to describe the relationship between elements should be interpreted in a like fashion (e.g., “between” versus “directly between,” “adjacent” versus “directly adjacent,” etc.). As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items.

[0036] Although the terms first, second, third, etc. may be used herein to describe various elements, components, regions, layers and / or sections, these elements, components, regions, layers and / or sections should not be limited by these terms. These terms may be only used to distinguish one element, component, region, layer or section from another region, layer, or section. Terms such as “first,” “second,” and other numerical terms when used herein do not imply a sequence or order unless clearly indicated by the context. Thus, a first element, component, region, layer, or section discussed below could be termed a second element, component, region, layer, or section without departing from the teachings of the example embodiments.

[0037] Spatially relative terms, such as “inner,” “outer,” “beneath,” “below,” “lower,” “above,” “upper,” and the like, 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. Spatially relative terms may be intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if the device in the FIG. is turned over, elements described as “below”, or “beneath” other elements or features would then be oriented “above” the other elements or features. Thus, the example term “below” can encompass both an orientation of above and below. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly.

[0038] As shown in FIG. 1, the portable sensor system 100 may be integrated into a conventional shipping container of a food sample and turn the container into a smart-monitoring device for the presence of pathogenic bacteria in the shipping container. As shown in FIG. 2, 6, and 8-10, the portable sensor system 100 may include a containment portion 102 and a sensor portion 104. The containment portion 102 may include a shipping container such as ajar. The sensorAttorney Docket No. 70673-02 portion 104 may include a cap 106 that may be selectively coupled with the containment portion 102. As shown in FIGS. 2-3, 11, the cap 106 may include a battery module 108, a processing module 110, a sensor module 112, and a coupling mechanism 114. In a specific example, the portable sensor system 100 may be provided in four layers. For instance, the battery module 108 may be provided as a battery layer, the processing module 110 may be provided as a processing layer, the sensor module 112 may be provided as a sensor layer, and the coupling mechanism 114 may be provided as a coupling layer. Each of the layers may be stacked and coupled together, thereby providing the cap 106. More particularly, each of the layers may be disposed in line with each other along the same axis. The coupling mechanism 114 may be configured to selectively lock the cap 106 to the containment portion 102. In a more specific example, one or more of the aforementioned layers may be 3D printed. In an even more specific example, two or more of the layers may be coupled together with machine screws binding them together. In certain circumstances, the coupling mechanism 114 may include a machine screw that may be selectively attached and / or detached by a user to selectively lock and unlock the cap 106 from the containment portion 102. In certain circumstances, all the electronics of the portable sensor system 100 may be disposed in the cap 106. Provided as a non-limited example, as shown in FIG. 12, the processing module 110 may consist of a processing unit 116, a voltage regulator 118, and / or a toggle button 120. The processing unit 116 may include an Arduino nano RP2040. The toggle button 120 may include a dual channel toggle button. The voltage regulator 118 may include two 5- volt voltage regulators. In certain circumstances, the processing unit 116 may perform the functions of using the battery module 108 and / or the sensor module 112 based on predetermined instructions. The sensor module 112 may include a silicon photomultiplier (SiPM) light sensor 122 and / or a verification light 124. The SiPM light sensor 122 may include a Hamamatsu C15524-1315SA silicon photomultiplier type light sensor module. As shown in FIGS. 5A-5B, the sensor module 112 may have comparable and / or enhanced performance compared to a standard lab luminometer. The verification light 124 may include an LED and two rechargeable 9-volt batteries. A portion of an outer surface of the portable sensor system 100 may include a sheath 126 that holds the containment portion 102 in alignment with the sensor module 112 and may block exterior light from interfering with measurements made. In other words, the sheath 126 may at least partially cover the containment portion 102 and militate against light from entering the containment portion 102. In an even more specific example, the portable sensor system 100 may use the two 9-volt batteries whichAttorney Docket No. 70673-02 are wired to + / - 5-volt regulators to produce positive and negative 5-volt potentials for the analog SiPM sensor 122. One battery may be used to power the processing unit 116. The portable sensor system 100 may include a data access port 123, such as a micro-USB port, to access files where data is stored. The LED 124 may be partially obscured near the SiPM sensor 122 which may automatically light up as a test to ensure that the SiPM sensor 122 is working properly. One skilled in the art may select other suitable ways for providing the portable sensor system 100, within the scope of the present disclosure.

[0039] As a non-limiting example, the processing unit 116 may include a microcontroller. The microcontroller and the SiPM light sensor 122 may be integrated into the stand-alone cap 106, as shown in FIGS. 15-17, that replaces an original bottle cap of the sample container. The portable sensor system 100 may also include a reporter phage, which may then provide historical data log of the optical readout from the portable sensor system 100 throughout the whole shipping period to the third-party lab or government laboratories, as shown in FIG. 18. The historical data log may be retrieved from the portable sensor system 100 via the data access port 123. Advantageously, since these locations generally receive large quantities of samples to be tested, the portable sensor system 100 may provide a quick readout of which samples to be prioritized screened for a confirmatory study and subsequent intervention procedure.

[0040] It is further contemplated that different sensors and / or lens arrays may be implemented to improve the sensitivity of the portable sensor system 100. Using lens arrays and diffusive materials may only go so far to improve sensitivity and high exposure time camera sensors are also considered as a replacement to the SiPM light sensor 122. The ability to access each pixel of a camera individually may also give software the ability to filter out noise, adding another vector by which sensitivity can be improved. Additionally, a rechargeable battery or batteries may be installed in order to simplify ease of use.

[0041] The portable sensor system 100 may be provided in various ways. For instance, as shown in FIG. 19, the portable sensor system 100 may be used according to a method 200. The method 200 may include a step 202 of providing a sensor portion 104 and a containment portion 102. The sensor portion 104 may include a battery module 108, a processing module 110, and a sensor module 112. Next, a sample in the containment portion 102 may be collected to be tested for a target bacteria. Afterwards, a bacteriophage-based assay may be performed in the containment portion 102. Then, a lux gene may be introduced to the containment portion 102 via phage. Next, asAttorney Docket No. 70673-02 shown in FIG. 4, a light may be monitored from an expressed lux gene if a target bacteria is present in the sample. In certain circumstances, the method 200 may also include saving a historical data log of the bacteriophage-based assay. The historical data log may then be retrieved from a data access port 123 on the portable sensor system 100. In another specific example, a sheath 126 may be disposed at least partially over the containment portion 102. A skilled artisan may select other suitable methodologies for using the portable sensor system 100, within the scope of the present disclosure.

[0042] Advantageously, the portable sensor system 100 of the present disclosure provides an in-transit bioluminescence-based pathogen monitoring system. The portable sensor system 100 may utilize a silicon photomultiplier sensor 122 to detect photons from a target assay. The portable sensor system 100 may be manufactured with 3D printing. The battery module 108, the processing module 110, the sensor module 112, and the coupling mechanism 114 may be integrated into a format of a single bottle cap 106 mounted on conventional sampling bottles.

[0043] Example embodiments are provided so that this disclosure will be thorough and will fully convey the scope to those who are skilled in the art. Numerous specific details are set forth such as examples of specific components, devices, and methods, to provide a thorough understanding of embodiments of the present disclosure. It will be apparent to those skilled in the art that specific details need not be employed, that example embodiments may be embodied in many different forms, and that neither should be construed to limit the scope of the disclosure. In some example embodiments, well-known processes, well-known device structures, and well-known technologies are not described in detail. Equivalent changes, modifications and variations of some embodiments, materials, compositions, and methods can be made within the scope of the present technology, with substantially similar results.

Claims

Attorney Docket No. 70673-02CLAIMSWHAT IS CLAIMED IS:

1. A portable sensor system configured to detect the presence of a pathogenic bacteria, wherein the system comprises: a sensor portion provided as a cap which includes a battery module, a processing module, a sensor module, and a coupling mechanism.

2. The system of Claim 1, further comprising a containment portion selectively coupled to the sensor portion.

3. The sensor of Claim 2, wherein the containment portion is ajar.

4. The sensor of Claim 1, wherein the processing module includes a processor and a voltage regulator.

5. The sensor of Claim 1, wherein the sensor module includes at least one of a high exposure time camera sensor, a lens array, and a silicon photomultiplier light sensor.

6. The sensor of Claim 1, wherein the sensor module includes a silicon photomultiplier light sensor that detects photons from a target assay.

7. The sensor of Claim 1, wherein the sensor module includes a verification light.Attorney Docket No. 70673-028. The sensor of Claim 2, wherein the containment portion is at least partially covered by a sheath that militates against light from entering the containment portion.

9. The sensor of Claim 1, further comprising a reporter phage.

10. The sensor of Claim 1, wherein the sensor portion includes a data access port.

11. The sensor of Claim 1 , wherein the battery module is a provided as a battery layer, the processing module is provided as a processing layer, the sensor module is provided as a sensor layer, the coupling mechanism is provided as a coupling layer, and each of the battery layer, the processing layer, the sensor layer, and the coupling layer are disposed in line with each other along the same axis.

12. The sensor of Claim 11, wherein each of the battery layer, the processing layer, the sensor layer, and the coupling layer are three-dimensionally printed.

13. The sensor of Claim 1, wherein the battery module includes rechargeable batteries.

14. A method of using a portable sensor system, the method comprising the steps of: providing a sensor portion and a containment portion, the sensor portion including a battery, a processing module, and a sensor module; collecting a sample in the containment portion to be tested for a target bacteria; performing a bacteriophage-based assay in the containment portion; introducing a lux gene via phage; and monitoring for a light from an expressed lux gene if a target bacteria is present in the sample.Attorney Docket No. 70673-0215. The method of Claim 14, further comprising a step of saving a historical data log of the bacteriophage-based assay.

16. The method of Claim 15, further comprising a step of retrieving the historical data log from a data access port on the portable sensor system.

17. The method of Claim 14, further comprising a step of disposing a sheath at least partially over the containment portion.