Automatic system for preparing and analyzing a biologic sample and method thereof

Abstract

The disclosure relates to a tube and automatic system for preparing and analyzing a biologic sample, such as a stool sample. The tube comprises a housing with an upper portion featuring a cone and a sealing element to receive a sample collector stick. A dispensing valve in the lower portion allows fluid exit to facilitate sample dilution and dispensing. Upon insertion of the sample collector stick, the tube enables a pressure differential to control fluid movement. The system includes a mixing mechanism for sample dilution, utilizing an external vibrator and optional solid elements for efficient mixing. The automatic system coordinates preparation and analyte detection, featuring a container housing with chambers for a test cassette and the tube. A controller operates mixing, valve actuation, and image capture of the test cassette, optionally communicating with remote devices. The system is adaptable for multiple diagnostic tests involving analyte concentration detection, with components to align and secure orientations within the automated system. Methods for using these components are disclosed.

Description

AUTOMATIC SYSTEM FOR PREPARING AND ANALYZING A BIOLOGIC SAMPLE AND METHOD THEREOFFIELD OF THE INVENTION

[0001] The present invention is in the field of sample devices and methods for preparing and detecting biologic samples.BACKGROUND OF THE INVENTION

[0002] Early diagnosis of gastrointestinal disorders is paramount given the rising prevalence of such conditions, including inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), infections, and gastrointestinal cancer. Stool samples are commonly utilized for diagnostic purposes owing to their accessibility and diagnostic potential. However, the efficacy of rapid tests in home-based and / or point of care for stool sample analysis is hindered by challenges related to user reluctance, and discomfort in handling such samples, resulting in user non-compliance, as well as perceived technical complexity.

[0003] For example, it is known that the concentration of the neutrophil-derived marker, fecal calprotectin (FC) reflects mucosal healing in IBD and is associated with endoscopic activity scales. Increased FC levels have been shown to correlate with IBD activity. The concentration of FC in stool samples is usually measured by quantitative enzyme-linked immunosorbent assay (FC-ELISA). However, this method necessitates a well-equipped laboratory to analyze the samples, accompanied with high financial cost, and slow evaluation time. FC home tests measure FC levels by immunochromatographic method using red and gold particles as rising through a test cassette and reaching testing and control strips.

[0004] Nonetheless, FC home tests still require user handling with feces; the user must manually sample their stool in at least three different places using a collection stick to ensure homogeneous sampling. Then, the user is requested to insert the collection stick into a dilution tube and thoroughly shake it to ensure the sample is adequately diluted within the solution. The user is then required to load the diluted sample onto the test cassette. Finally, the user is requested to take an image of the results window of the test cassette at an accurate timing, after calibration process, which is then analyzed by a smartphone application. The reliance on the user's sampling method, manual mixing, loading, and ability to capture an image after a predefined time in a non-controlled illuminating environment, all lead to significant variance in results, as well as raise the chance for sample contamination derived from user manual handling of the sample.

[0005] Indeed, it has been demonstrated that results obtained using these FC home tests are less accurate compared to FC-ELISA (Lerchova, Tereza; Hradsky, Ondrej; Copova, Ivana; Potuznikova, Kristyna; Gonsorcikova, Lucie; Bronsky, Jiri. The Accuracy of a Home- performed Faecal Calprotectin Test in Paediatric Patients With Inflammatory Bowel Disease. Journal of Pediatric Gastroenterology and Nutrition 69(1): p 75-81, 2019).

[0006] Moreover, a prospective study examining FC test performed at home by parents of pediatric patients with IBD, revealed that 15% of the home-tests failed, mainly because of technical reasons, including technical problems with the test (77%), problems with smartphone camera (13%), and inappropriate storage of the tests. Just under half of the patients (47%) considered home testing comparable or superior to routine testing. Most of the parents were unsatisfied (61%), since the results were significantly different from FC-ELISA and due to technical issues derived from handling phone application. Interestingly, when the home test was performed by a laboratory professional it was comparable with ELISA, suggesting that practical issues hampered home testing (Piekkala M, Alfthan H, Merras-Salmio L, Puustinen Wikstrbm A, Heiskanen K, Jaakkola T, Klemetti P, Farkkila M, Kolho KL. Fecal Calprotectin Test Performed at Home: A Prospective Study of Pediatric Patients With Inflammatory Bowel Disease. J Pediatr Gastroenterol Nutr. 2018; 66:926-931).

[0007] Therefore, there is an urgent need for new automatic methods to prepare and accurately analyze analytes in biological samples, such as measuring FC levels in stool samples.SUMMARY OF THE INVENTION

[0008] It is an object of the present invention to disclose a tube for receiving a biologic sample, and dispensing a predetermined volume of diluted the biologic sample, comprising: a tube housing configured to accommodate fluid, the tube housing having an upper portion and a lower portion; the upper portion comprises a cone comprising an upper opening and a hole at the lower end of the cone, the hole is provided with a temporally central sealing element; the upper portion is configured for receiving a sample collector stick comprising a longitudinal segment with collector gripping segment and a lower sample collecting segment configured to accommodate a biologic sample; the opening of the tube lower portion comprises a dispensing valve, configured in a closed position preventing the fluid from exiting from the opening of the lower portion, and operable in an open position to permit the fluid to exit; wherein, once the collector stick is inserted through the hole to a predetermined position, the dimensions of the cone and the hole are configured to create a seal with the longitudinalsegment of the collector stick, generating a predetermined increased pressure (AP) of air above the fluid, further wherein, the longitudinal segment in the predetermined position comprises an internal segment and an external segment, and the dimensions of the internal segment are proportional to the predetermined volume of the fluid exiting from the lower portion, for generating dispensing of a predetermined volume of the fluid once the valve is in the open position.

[0009] It is another object of the present invention to disclose a tube as disclosed herein, wherein the predetermined position of the collector stick within the tube is when the lower sample collecting segment extends into the fluid, the gripping segment closes the upper opening, or both.

[0010] It is another obj ect of the present invention to disclose a tube as disclosed herein, further comprising a ring in the upper opening, and the collector gripping segment comprises a sample collector locking element, configured to lock the ring, when the collector stick is in the predetermined position.[Oi l] It is another object of the present invention to disclose a tube as disclosed herein, wherein the cone is a press-fitted cone.

[0012] It is another object of the present invention to disclose a tube as disclosed herein, further comprising an elastomeric ring at the lower end of the cone, and the hole is a cavity within the elastomeric ring.

[0013] It is another object of the present invention to disclose a tube as disclosed herein, wherein the central sealing element is a silicone plug or an elastomeric analogue thereof.

[0014] It is another object of the present invention to disclose a tube as disclosed herein, wherein central sealing element is pressed into the fluid upon insertion of the sample collector stick through the hole.

[0015] It is another obj ect of the present invention to disclose a tube as disclosed herein, further comprising a mixing mechanism, configured to homogenously dilute the biologic sample loaded on the collecting segment, within the fluid, and the dispensing is of a predetermined volume of a homogenous mixture of the fluid and the sample.

[0016] It is another object of the present invention to disclose a tube as disclosed herein, wherein the mixing mechanism is vibrations by an external automatic vibrator.

[0017] It is another obj ect of the present invention to disclose a tube as disclosed herein, further comprising at least one solid element within the fluid, configured to vibrate when the external automatic vibrator is operated, to facilitate and / or shorten the time required to homogeneously dilute the biological sample loaded onto the collecting segment in the fluid.

[0018] It is another object of the present invention to disclose a tube as disclosed herein, wherein the at least one solid element has a density lower than 1 gr / ml.

[0019] It is another obj ect of the present invention to disclose a tube as disclosed herein, further comprising a lateral external longitudinal projection configured to: a. restrict rotational movement of the tube within a cylinder chamber of an automatic system for preparing and analyzing the biologic sample, the automatic system comprises a valve actuator; and, b. place the valve in adjacent to the valve actuator.

[0020] It is another object of the present invention to disclose a tube as disclosed herein, wherein the ratio (v / v) between the volume of the internal segment of the sample collector stick, and the predetermined volume of the fluid is between 5: 1 to 1 :5 in room temperature.

[0021] It is another object of the present invention to disclose a tube as disclosed herein, wherein the ratio is about 1 : 1.

[0022] It is another object of the present invention to disclose a tube as disclosed herein, wherein the biologic sample is a stool sample.

[0023] It is another object of the present invention to disclose an automatic system for preparing diluted biologic sample and detecting an analyte in the biologic sample, comprising: a container housing; a first chamber configured for receiving a test cassette of a biologic analyte; a second chamber configured for receiving a tube, the tube is configured for receiving a sample collector stick comprising a biologic sample, the tube comprises an upper portion comprising a cone comprising an upper opening and a hole at the lower end of the cone, the hole is provided with a temporally central sealing element, the opening of the tube lower portion comprises a dispensing valve, configured in a closed position preventing the fluid from exiting from the opening of the lower portion, and operable in an open position to permit the fluid to exit; at least one controller, operably linked to: a. a mixing mechanism configured to homogenously dilute the biologic sample within the fluid; b. a valve actuator configured to open the valve after the mixing and to dispense a predetermined volume of homogenous mixture; c. illumination means and camera configured to capture an image of the test cassette; and, d. optionally user display on the container housing,wherein the at least one controller comprises instructions to activate the mixing mechanism for a first predetermined time, to further automatically activate the valve actuator to open the valve and to dispense a predetermined volume of the homogenous mixture onto the test cassette, and to further automatically operate the illumination means and camera to capture an image of the test cassette after a second predetermined time, to produce an automatic preparation and detection of the biologic sample.

[0024] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the test cassette is configured to produce color development proportional to concentration of the biologic analyte in the biologic sample, and the illumination means and camera are configured to capture an image of the color development within the test cassette.

[0025] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein at least one controller is operably linked to a remote operator via communication means, the remote operator is configured to perform at least one member of the group consisting of: a. analyzing the image and / or image based-data; b. storing at least one member of the group consisting of the image, the image based-data and analysis of the image based-data; and, c. displaying at least one member of the group consisting of the image, the image based-data and analysis of the image based-data in at least one of a user's smartphone, a medical data system, or analogue system thereof.

[0026] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the remote operator is a cloud-based system, further comprising data of a standard curve of the analyte, for a specific batch of the test cassette, and analyzing of the image and / or image based-data is based on the standard curve.

[0027] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein at least one controller is operated by the remote operator such as the user's smartphone, or analogue device thereof.

[0028] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the test cassette is a lateral flow cassette.

[0029] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the lateral flow cassette comprises a sample loading area and a test results window.

[0030] It is another object of the present invention to disclose an automatic system as disclosed herein, wherein the second chamber is positioned at a first predetermined distance above thefirst chamber, to allow optimal liquid dripping and accurate absorption of the predetermined volume by the sample loading area, once the tube is in the second chamber.

[0031] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the illumination and camera means are located at a second predetermined distance above the test result window.

[0032] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, further comprising at least one sensor operably linked to the at least one controller, the at least one sensor detects at least one member of the group consisting of the presence, the orientation, the type, or any combination thereof, of the test cassette, the presence, orientation, or both, of the tube, the presence of the sample collector stick within the tube, the presence and / or dissolution of the sample within the buffer, and any combination thereof.

[0033] It is another object of the present invention to disclose an automatic system as disclosed herein, further comprising a sensor configured to detect the dissolution of the sample within the buffer, and the first predetermined time is when the dissolution of the sample within the buffer is above a predetermined threshold, creating a homogenous solution.

[0034] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, further comprising a lid, wherein closure of the lid enables at least one member of the group consisting of noise reduction, minimal user contact, a complete sealing of the system from external light, or any combination thereof.

[0035] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the valve actuator is connected to a valve connector.

[0036] It is another object of the present invention to disclose an automatic system as disclosed herein, wherein the valve actuator comprises a mechanical interface mean that enables engagement to the valve connector and a rotational movement of the valve connector to switch the valve from the closed position to the open position.

[0037] It is another object of the present invention to disclose an automatic system as disclosed herein, further comprising a longitudinal notch, compatible to an external longitudinal projection positioned alongside the tube, the notch is configured to mark insertion site for a user, by restricting rotational orientation of the tube, allowing position of the valve connector adjacent to the valve.

[0038] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, further comprising a flexible ring surface, on or adjacent to the upper side of the container housing, configured to encircle the tube and / or the collector gripping segment and to allow vibrations of the tube, once the mixing mechanism operates.

[0039] It is another object of the present invention to disclose an automatic system as disclosed herein, wherein the mixing mechanism comprises vibrations by a vibrator, coupled to the second chamber.

[0040] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the vibrator is positioned vertically and coupled to a bottom portion of the second chamber.

[0041] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the biologic sample is a stool sample.

[0042] It is another obj ect of the present invention to disclose an automatic system as disclosed herein, wherein the biologic analyte is a protein or nucleic acid whose levels are dysregulated in a gastrointestinal disease selected from the group consisting of: gastrointestinal cancer, gastrointestinal infection and inflammatory bowel disease (IBD).

[0043] It is another object of the present invention to disclose an automatic system as disclosed herein, wherein the test cassette is selected from the group consisting of calprotectin test, fecal occult blood test (FOBT), fecal immunochemical test (FIT), stool bacterial culture, Clostridium difficile (C. difficile) toxin test, c-reactive protein (CRP), rotavirus and / or norovirus antigen tests, parasite tests, Helicobacter pylori (H. pylori) and any combination thereof.

[0044] It is another object of the present invention to disclose a method for preparing diluted biologic sample and detecting an analyte in the biologic sample, the method comprising: a. obtaining a sample collector stick comprising a biologic sample within a sample collecting segment; b. obtaining a tube comprising: fluid; an upper portion; and a lower portion, the upper portion comprises an upper opening and a hole provided with a temporally central sealing element; the opening of the tube lower portion comprises a dispensing valve, configured in a closed position preventing the fluid from exiting from the opening of the lower portion, and operable in an open position to permit the fluid to exit; c. inserting the sample collector stick into the tube to a predetermined position via the hole; d. obtaining an automatic system comprising: a first chamber configured for receiving a test cassette of a biologic analyte, the test cassette is configured to produce color development proportional to concentration of the biologic; a second chamber configured for receiving the tube; at least one controller, operably linked to:i. a mixing mechanism configured to prepare a homogenous mixture of the biologic sample and the fluid, within the tube; ii. a valve actuator configured to open the valve after the mixing and to dispense a predetermined volume of the homogenous mixture; iii. illumination means and camera configured to capture the color development within the test cassette; and iv. optionally user display on the container housing, wherein the at least one controller comprises instructions to activate the mixing mechanism for a first predetermined time, to further automatically activate the valve actuator to open the valve and to dispense a predetermined volume of the homogenous mixture onto the test cassette, and to further automatically operate the illumination means and camera to capture the test cassette after a second predetermined time, to produce an automatic preparation and analysis of the biologic sample; e. turning on the automatic system; f. inserting a lateral flow cassette into the test cassette chamber, inserting the tube comprising the sample collector stick into the second chamber; and, g. obtaining at least one member of the group consisting of an image, an image-based data, and analysis of the image-based data, in at least one of user's smartphone, a medical data system, or a corresponding system thereof.

[0045] It is another object of the present invention to disclose a method as disclosed herein, wherein the step (f) comprises fully inserting the tube comprising the sample collector stick into the second chamber, allowing the valve actuator to be positioned adjacent to the valve.

[0046] It is another object of the present invention to disclose a method as disclosed herein, further comprising at least one sensor operably linked to the at least one controller, the at least one sensor detects at least one member of the group consisting of the presence, the orientation, the type, or any combination thereof, of the test cassette, the presence, orientation, or both, of the tube, the presence of the sample collector stick within the tube, the presence and / or dissolution of the sample within the buffer, and any combination thereof.

[0047] It is another object of the present invention to disclose a method as disclosed herein, wherein the user's smartphone or analogue device thereof displays at least one notification that validates the at least one member of the group consisting of the presence, the orientation, the type, or any combination thereof, of the test cassette, the presence, orientation, or both, of the tube, the presence of the sample collector stick within the tube , the presence and / or dissolution of the sample within the buffer, and any combination thereof.

[0048] It is another object of the present invention to disclose a method as disclosed herein, wherein the biologic sample is a stool sample and the step (a) further comprises collecting the stool sample on the sample collecting segment by a stool sampling device configured to accommodate the sample collector stick, the stool sampling device comprises a proximal wiping base and a distal covering element or portion, configured to sample a predetermined amount of the stool on the collecting segment.

[0049] It is another object of the present invention to disclose a method as disclosed herein, wherein collecting the stool sample comprises a manual sampling by contacting the stool sampling device with at least one portion of the stool.

[0050] It is another object of the present invention to disclose a method as disclosed herein, wherein the stool sampling device is accommodated within a stool collecting device configured to collect a homogenous stool sample comprising a stool receiving container; and a lid for sealing the container, and the collecting the stool sample comprises rotating the lid to obtain the homogenous stool sample.

[0051] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0052] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The presently disclosed subject matter may be more clearly understood upon reading of the following detailed description embodiments of non-limiting exemplary embodiments thereof, with reference to the drawings.

[0054] The following detailed description of embodiments of the presently disclosed subject matter refers to accompanying drawings. Dimensions of components and features shown in figures are chosen for convenience or clarity of presentations and are not necessarily shown to scale. Wherever possible, the same reference numbers will be used throughout the drawings and the following description to refer to the same and like parts.

[0055] Figures 1A-1B are illustrations representing a perspective view of a tube 100 including a valve in a close configuration (1A) and open configuration (IB) according to some embodiments of the invention.

[0056] Figures 2A-2C are illustrations representing 3D perspective internal view of a tube 100, including a press-fitted cone 111 and a round-shaped valve 160.

[0057] Figure 3 is an illustration representing a perspective view a tube 100 and a sample collector stick 200 according to some embodiments of the invention. Insertion of the sample collector stick into the tube pushes a central sealing element, e.g., plug 112 temporarily sealing the tube into the fluid within the tube.

[0058] Figure 4 is a schematic presentation of the system 300 disclosed herein, according to some embodiments of the invention, comprising the tube 100, sealed in its upper portion by the sample collector stick 200.

[0059] Figures 5A-5B are illustrations representing a perspective view of the external view of the system 300 disclosed herein according to some embodiments of the invention.

[0060] Figures 6-10 are illustrations representing a perspective view of the internal view of the system 300 disclosed herein according to some embodiments of the invention. Figure 6 is an illustration of tube 100 comprising the sample collector stick 200, within second chamber 323. Figures 7A-7B show the valve actuator 351, valve connector 350, and the mixing mechanism 340, according to some embodiments of the invention. Figure 8 shows major features within the system 300 disclosed herein, including the position of the tube 100 in relation to the flow cassette 380, and the position of the illumination means and camera 360 in relation to the flow cassette 380. Figures 9A-9B and Figures 10A-10B further demonstrate some of the components of the system disclosed herein, according to some embodiments of the present invention including communication means 370.

[0061] Figure 11 is a flow chart exhibiting the method disclosed herein, according to some embodiments of the invention.

[0062] Figures 12A-12E demonstrate an exemplary stool sampling device 400 designed to accommodate the sample collector stick 200 when collecting a stool sample.

[0063] Figure 13 illustrates an exemplary stool collecting device 500 configured to collect a homogenous stool sample comprising a stool receiving container 501 and a lid 502.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0064] The present invention is based at least in part, on the surprising finding that biologic analytes, such as fecal calprotectin (FC) levels, can be automatically and accurately measured using the tube and the system disclosed herein. The present invention is further based on the finding that preparation of homogenous mixture of stool samples diluted in a buffer, and automatic dispensing of a pre-determined volume of the homogenous mixture can be performed by the tube disclosed herein.

[0065] Fecal calprotectin (FC) levels can be used as a quantitively biomarker, for example for distinguishing between IBD and IBS, for detecting relapses of IBD and for monitoring effectivity of IBD therapies. In patients with Crohn’s disease (CD) in symptomatic remission with recent confirmation of endoscopic remission, the American Gastroenterological Association (AGA) suggests using fecal calprotectin <150 mg / g to rule out active inflammation (Ananthakrishnan A.N., Adler J., Chachu K.A., et al, AGA Clinical Practice Guideline on the Role of Biomarkers for the Management of Crohn’s Disease, Gastroenterology 2023;165:1367-1399). In clinical trials where a response to a new treatment is monitored, a low cut-off point of 100 pg / g is frequently used to demarcate the upper limit of the normal FC range. Conversely, in real-life studies, a higher cut-off point of e.g. 250 pg / g is advocated as an action threshold for adjusting treatment (Maaser C., et al, ECCO-ESGAR Guideline for Diagnostic Assessment in IBD Part 1 : Initial diagnosis, monitoring of known IBD, detection of complications, Journal of Crohn's and Colitis, Volume 13, Issue 2, February 2019, Pages 144-164).

[0066] The need for an automatic device, with minimal user interaction with a biologic sample (e.g., a stool sample) and minimal steps performed by the user, is necessary. However, this should not undermine the necessity of overcoming challenges in analyzing a quantitatively biomarker whose levels indicate the presence and / or the severity of a disease. The system disclosed herein allows for homogenized sampling of a biologic sample, such as stool, with minimal user contact. The sample is collected and transferred to the automatic device at a predetermined volume or weight to prevent false-positive or false-negative results.Furthermore, the buffer-diluted sample is automatically mixed, without relying on user capabilities. A predetermined volume of the homogeneous buffer-diluted sample is then automatically loaded onto a lateral flow cassette. The system then captures an image of the test results window after a predetermined time, compatible with the examined test and the specific lateral cassette, and results are finally analyzed. Only adaptation of all required parameters enables a sensitive test that is both easy to use without leading to user noncompliance and yet provides accurate results.

[0067] Reference is now made to Figure 1, Figures 2A-2C, and Figure 3, demonstrating exemplary illustrations of a tube 100 as disclosed herein. Tube 100 is configured a) to receive a biologic sample placed onto a sample collecting segment 220 within a sample collector stick 200; b) to dilute the biological sample within a buffer to produce a homogenous solution, wherein all the collected biologic sample is homogenously dissolved within the buffer; and, c) to dispense or extract a predetermined volume of the buffer-diluted sample.

[0068] In some embodiments, the biological sample is selected from the group consisting of stool, whole blood, serum, plasma, cerebrospinal fluid, spinal fluid, amniotic fluid, aqueous humor, vitreous humor, bile, breast milk, cerumen (earwax), chyle, chyme, endolymph, perilymph, exudates, female ejaculate, gastric acid, gastric juice, lymph, mucus (including nasal drainage and phlegm), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheum, saliva, sebum (skin oil), semen, sputum, synovial fluid, sweat, tears, urine, vaginal secretion, vomit and mixtures of one or more thereof. In some embodiments, the biologic sample comprises stool sample. The stool sample collected on the sample collecting segment 220 may be in any aggregation state selected from liquid, solid, or any state in-between.

[0069] In some embodiments, the tube housing 110 is a cylindrical container. In some embodiments, the tube housing 110 is made of a biocompatible durable material such as plastic or glass. In some embodiments, the tube is made of disposed material. The tube 100 is preferably composed of a material that is resilient to withstand increased pressure exerted by the inserted sample collector stick 200 without deforming or rupturing. Non limiting examples for such a material include plastic, silicone, thermoplastic elastomers (TPEs), and certain types of rubber. In some embodiments, tube 100 is made from materials selected from polypropylene (PP), polycarbonate (PC), polyethylene (PE), polyethylene terephthalate (PET), glass, and any combination thereof.

[0070] Still referring to Figure 1 and Figures 2A-2B, the tube 100 has an upper portion 130 and a lower portion 140. In the exemplified example, the upper portion 130 of the tube comprises a ring 132 circulating the upper opening 131 (Figure 2B). In some embodiments, the ring 132 is made of flexible material. In some embodiments, below the ring 132 level, there is positioned a cone 111, comprising an upper opening 116 and a hole 113 at the lower end of the cone 111. In some embodiments, cone 111 comprises an elastomeric ring 115 at the lower end of cone 111, and the hole 113 is positioned within the elastomeric ring 115.

[0071] The sample collector stick 200, as exemplified in Figure 3, is configured to be inserted through hole 113. The shape of a cone 111, in some embodiments, is configured to facilitate or direct insertion of the sample collector stick 200 into the tube, and to ensure removal of excessive sample collected on the sample collector stick 200.

[0072] In some embodiments, hole 113 is temporarily sealed with a central sealing element 112. In some embodiments, cone 111 is a press-fitted cone, pressed into the tube. In some embodiments, the collector gripping segment 230 comprises a locking mean 240, configured to lock the ring 132, and secure the collector stick in place when it is fully inserted.

[0073] As used herein, the term "central sealing element" refers to any element or mean that can at least temporarily seal the hole 113 within the upper portion of the tube. In some embodiments, the central sealing element is selected from the group consisting of: a plug, a stopper, a cap, a lid, a gasket, and any combination thereof. In some embodiments, the central sealing element is a press-fitted element.

[0074] In some embodiments, the central sealing element 112 is a plug made of a flexible material, such as silicone, allowing for a sealed closure of the hole 113. The diameter of hole 113 is designed to fit that of the longitudinal segment 210 of the sample collector stick 200, allowing for a sealed closure once the sample collector stick 200 is within the tube 100,

[0075] In some embodiments, the fitted diameter of hole 113 allows for the removal of excessive sample present on the sample collecting segment 220 once the sample collector stick 200 is inserted into hole 113, enabling a predetermined ratio between the sample and buffer with the tube 200.

[0076] As used herein, the term "elastomeric ring" encompasses a circular or ring-shaped component made from elastomeric materials such as rubber or silicone.

[0077] Reference is still made to Figures 1A-1B, showing the lower portion 140 of the tube 100. Below the fluid or buffer level 120, there is a first lower opening 141 on the proximal side of the fluid 120. The first lower opening 141 is sealed by a valve 160, which is placed in a closed position 161, preventing fluid exit, and can be operated in an open position 162,permitting fluid to exit. In some embodiments, a canal or a conduit connects the first lower opening 141 and the second lower opening 142, positioned distally to the fluid level, from which the fluid is dispensed out of the tube. The valve, in its closed configuration 161, blocks the transmission of the fluid 120 via the canal.

[0078] In some embodiments, valve 160 is a cone-shaped valve. As demonstrated in Figures 2B and 2C, in some embodiments, the internal portion of the cone present within the tube is round-shaped, preventing the deposition of aggregates that may be present within the solution onto the valve. This design thus prevents the possible blockade of channel 141 once the valve is transformed to its open position 162.

[0079] In some embodiments, the valve is held in a closed position 161 when the sample collector stick 200, containing the sample within the sample collecting segment 220, is inserted via hole 113 in the upper portion 130. Preferably, as described in Figure 3, column 4, the insertion of the sample collector stick 200 through the hole 113 to a predetermined location pushes the central sealing element, e.g., plug 112 into the fluid 120. The predetermined location can be when the collector gripping segment 230 closes the opening 131 of the tube 100, when the locking mean 240 locks the ring 132, when the sample collecting segment 220 is within the fluid or the buffer, and any combination thereof. In some embodiments, the sample collecting segment 220 is tapered and comprises an angled surface. In some embodiments, the diameter of the sample collecting segment 220 is lower compared to the diameter of the hole 113, allowing slidable insertion of the sample collector stick 220 via the hole 113.

[0080] Once inserted into a predetermined location, a sealed closure of the upper portion of the tube is created between hole 113 and the longitudinal segment 210 of the sample collector stick 200. Since the lower portion of the tube 140 is sealed by the valve in the closed configuration161, an increased predetermined pressure (AP) of the air gap 114 is created within the tube. At this point, the tube containing the sample collector and the biological sample is sealed in both the upper and lower portions by the sample collector stick 200 and the valve in its closed configuration 161, respectively, allowing for easy portability and mobility of the tube 100 with the sample collector stick 200 inside.

[0081] Reference is still made to Figure 3. The tube 100 disclosed herein allows for a predetermined volume to be extracted or dispensed once the valve is moved to its open position162. When the sample collector stick 200 is in the predetermined location (e.g., when the locking mean 240 locks the ring 132), two portions of the longitudinal segment 210 can bedistinguished: an external segment 212, representing the segment above the hole 113, and an internal segment 211, referring to the segment within the tube, above the sample collecting segment 220.

[0082] When the sample collector stick 200 is in the predetermined location, the volume of the fluid dispensed out of the second lower opening 142 of the tube 100, by changing the configuration of the valve from a close position 161 to an open position 162, is proportional to the volume of the internal segment 211 of the sample collecting segment 220. In some embodiments, the longitudinal segment 210 has a cylinder shape, characterized by radius r, Hi for the height of internal segment 211 and H2 for the height of external segment 212. In some embodiments, the value of (nr2x Hi) is proportional to the volume of the dispensed fluid. In some embodiment, the ratio (v / v) between the volume of the internal segment 211 (nr2x Hi) and the volume of the dispensed fluid is between 5 : 1 to 1 :5 in room temperature (RT). In some embodiments, the tube disclosed herein allows a dispensing mechanism of a predetermined volume of fluid, wherein the ratio (v / v) between the volume of the internal segment 211 (nr2x Hi) and the volume of the dispensed fluid is between 2: 1 to 1 :2 in RT. In some embodiments, the ratio (v / v) is about 1 : 1 in RT.

[0083] In some embodiments, the predetermined volume is determined by predetermining the size, e.g., length of the cone 111. In some embodiments, the predetermined volume is determined by predetermining the position of the cone 111. In some embodiments, the predetermined volume is determined by predetermining the length, the radius or both of the longitudinal segment 210 and / or the internal segment 211 of the sample collector stick 200. In some embodiments, the predetermined volume is determined by the level of the buffer.

[0084] In some embodiments, an automatic mixing mechanism 340 is coupled to the tube 100 to dilute the biological sample within the fluid or buffer, allowing for homogeneous dilution of the sample.

[0085] As used herein, the term "mixing mechanism" refers to a device used to dissolve, blend, or combine, substances to achieve a uniform mixture. Non-limiting examples include vortex, magnetic stirring, vibrator, and ultrasonic mixing devices that use ultrasound to create cavitation and microstreaming in a liquid sample.

[0086] The mixing mechanism may be achieved either by ultrasonic mixing or by mechanical mixing (e.g., vibrations). In some embodiments, mechanical mixing is more efficient in homogeneously dissolving the sample within the buffer. Interestingly, the mechanical mixingmechanism disclosed herein was found to be associated with significantly less foaming of the buffer, compared to manual mixing.

[0087] In some embodiments, the predetermined location of the sample collector stick 200 within the tube is when the upper level of the fluid or buffer is only slightly above the sample collecting segment 220 comprising the biological sample. This arrangement was found to allow for maximal vibrations of the fluid interface, contributing to the efficient mixing mechanism.

[0088] In some embodiments, the volume of the fluid or buffer within the tube 100 is between 1 ml and 10 ml. In some embodiments, the volume of the fluid or buffer within the tube 100 is about 5 ml. In some embodiments, the sample collecting segment 220 is designed to collect 1- 10 mg of biologic sample. In some embodiments, the sample collecting segment 220 is designed to collect about 5 mg of biologic sample.

[0089] In some embodiments, mixing occurs for a time range between 10 seconds and 10 minutes. In some embodiments, mixing is for 0.5-5 minutes. In some embodiments, mixing occurs for a time range between 10 seconds and 120 seconds, between 10 seconds and 60 seconds, between 20 seconds and 120 seconds, between 20 seconds and 60 seconds, between 30 seconds and 120 seconds, between 30 second and 60 seconds. Each possibility represents a separate embodiment of the present invention. In some embodiments, mixing is for less than about one minute. In some embodiments, the system further comprises a sensor configured to detect the dissolution of the sample within the buffer 120, and the mixing ends when dissolution of the sample within the buffer is above a predetermined threshold, resulting in a homogenous solution. After achieving a homogeneous solution of buffer comprising the biological sample, an external valve actuator 351 can operate the valve to dispense a predetermined volume of the diluted sample within the homogeneous solution, allowing for dispensing of a predetermined volume of the diluted sample.

[0090] According to some embodiments, the tube further comprises at least one solid element within the fluid, configured to vibrate when the mixing mechanism 340, e.g., external automatic vibrator, is operated, to facilitate and / or shorten the time required to homogeneously dilute the biological sample in the fluid.

[0091] As used herein, the term “solid element(s)” refers to one or more objects that are present within the tube 100, which may be solid or semi-solid in nature, including flexible or deformable materials such as gels or elastomers. These elements are configured to float, sink, or remain suspended within the fluid, depending on their density, shape, and materialproperties, in order to facilitate agitation and mixing of the biological sample during operation of the mixing mechanism 340.

[0092] In some embodiments, at least one solid element may be of a three-dimensional shape such as a sphere, cube, pyramid, or any irregular or angular geometry. In some embodiments, the shape may not be symmetrical in all dimensions.

[0093] In certain embodiments, the solid element(s) may be made of a material having a density lower than 1 g / cm3(or 1 g / ml), such as polypropylene (with an exemplary specific gravity of approximately 0.9), when the target sample, loaded on the sample collecting segment 220, resides in the upper portion of the fluid. Conversely, if the biological sample is located near the bottom of the tube, materials with a density greater than 1 g / cm3may be used to ensure contact with the sample during vibration.

[0094] According to some preferred embodiments, the floating element(s) dimensions are selected to ensure sufficient contact with the region containing the sample. In some embodiments, the total volume of solid elements may range from 0.01% to 0.2% of the internal volume of the tube. For example, in a tube with an internal diameter of 14 mm, the floating element dimensions may have a volume ranging from approximately 0.2 mm3to 3 mm3.

[0095] The number of solid elements may vary, and in some embodiments, multiple elements may be included such that they form a layer or concentration of elements at the region of the biological sample. This ensures effective agitation in the vicinity of the sample during vibration.

[0096] In further embodiments, the tube may contain a mixture of solid elements differing in one or more parameters, such as shape, size, material, or density, to enhance mixing efficiency and coverage across different sample positions within the fluid tube.

[0097] In some embodiments, the central sealing element 112, the solid element(s), or both, further comprises a metal element. In some embodiments, when the sample collector stick 200 is inserted through hole 113 to a predetermined location, it pushes both the plug and the metal element into the fluid 120. In some embodiments, the presence of the metal element within the fluid enables the use of a mixing mechanism based on magnetic force (e.g., magnetic bead and / or stirrer).

[0098] In some embodiments, the tube 100 comprises a lateral external longitudinal projection 150 configured to: a) restrict rotational movement of the tube within a receiving chamber, and b) position the valve 160 adjacent to an external valve actuator configured to operate the valve.The external longitudinal projection 150 may be positioned above the valve, as seen in Figure 2A, allowing for accurate configuration of the valve 160 in relation to the valve actuator 351.

[0099] Reference is now made to Figures 4-6, demonstrating exemplary illustrations of the system 300 disclosed herein. System 300 is configured for automatically preparing a homogenous dilution of the biologic sample and detecting an analyte in the biologic sample. The automatic system 300 comprises a container housing 320, preferably sealed from external light, allowing controlled lighting environment within the container.

[0100] As the system disclosed herein is configured, at least in some embodiments, to detect an analyte within a biological sample by capturing an image of an analyte-detection mean in which color (or fluorescence) develops in relation to analyte concentration (e.g., in a strip or a cassette), the presence of a controlled light environment within the container housing enables obtaining accurate results without the need for a pre-calibration procedure of illumination before each test. In some embodiments, the presence of a controlled light environment allows for limiting the need for performing a calibration procedure to only before a new type, a new batch, or both, of cassettes and / or strips that have not been previously used in the system disclosed herein.

[0101] In some embodiments, the system 300 comprises at least two chambers; a) a first chamber 325 (as demonstrated in Figure 5B) is configured for receiving a test cassette or a strip 380 of a biologic analyte; and, b) a second chamber 323 (as demonstrated in Figure 6) is configured for receiving the tube 100 as disclosed herein. In some embodiments, the tube 100 is pre-sealed in its upper portion with the sample collector stick 200. In some embodiments, the second chamber 323 is a cylinder-shaped chamber, configured to wrap the tube 100.

[0102] The first chamber 325, configured for accommodating a test cassette or a strip, is preferably positioned at the lower portion of the system 300 and is accessed from the lateral side of the container housing 320. The test cassette or strip 380 is preferably positioned horizontally, although it may also be placed vertically or at any angle between the horizontal and vertical axes.

[0103] As exemplified in Figure 5B, optionally, chamber 325 is laterally opened by the user via a side access door. The second chamber 323 is preferably accessed via the upper portion of the container housing 320.

[0104] In some embodiments, the test cassette 380 is configured to produce color development proportional to the concentration of the biologic analyte in the biologic sample. The test cassette 380 may be a lateral flow cassette or a strip.

[0105] As used herein, the terms "a test cassette" and "strip" encompass a diagnostic device used for rapid testing of analytes (such as proteins, antibodies, hormones, drugs, etc.) in biological samples. The test cassette typically consists of a plastic cassette or strip with specific regions containing reagents or materials that interact with the analyte of interest, producing a detectable signal such as color change (i.e., colorimetric test) or fluorescence. The color or fluorescence development may be based on antigen-antibody interactions, enzyme-linked assays, chemical reactions triggered by the presence of the analyte, fluorescent dyes binding to the analyte, or any combination thereof. The test performed within the test cassette or strip may be selected from immunochromatographic assay, enzyme immunoassays (EIAs), enzyme- linked immunosorbent assay (ELISA), chemical reaction-based assay, immunochemical assay, polymerase chain reaction (PCR), immunoturbidimetric assay, immunonephelometric assay, and any combination thereof.

[0106] In some embodiments the test cassette 380 comprises a sample loading area 382 and a test results window 381.

[0107] The system 300 disclosed herein further comprises at least one controller or processor 330, operably linked to: a. a mixing mechanism 340 configured to homogenously dilute the biologic sample within the fluid 20 or the buffer; b. a valve actuator 351 configured to open the valve 160 after the mixing and to dispense a predetermined volume of homogenous mixture; c. illumination means and camera 360 configured to capture an image of the test cassette 380; and, d. optionally user display 327 on the container housing 320

[0108] As illustrated in Figure 4 and Figure 5A, the system 300 may further comprise a lid 310, wherein closure of the lid 310 enables at least one member of the group consisting of noise reduction, minimal user contact, a complete sealing of the system from external light, or any combination thereof.

[0109] Reference is still made to Figures 5A and 5B. A longitudinal notch or groove 322, compatible to the external longitudinal projection 150 of the tube 100 may be present alongside the internal portion of the second chamber 323, marking the insertion site of the tube 100 for the user and / or restricting rotational orientation of the tube 100 within the second chamber 323. In some embodiments, an accurate rotational position of the tube 100 within the second chamber 323 is required to allow the accurate position of the valve actuator 351, the valve connector 350, or both in regard to the valve 160 sealing the lower portion of the tube 100.

[0110] In some embodiments, once the user places the lateral flow cassette 380 into the test cassette chamber 325, inserts the sealed tube 100 pre-comprising the sample collector stick 200 into the second cylinder chamber 323, prefarably closes a lid 310, and turn on the system 300, a series of automatic steps is instructed by the controller 330.

[0111] Although the system 300 disclosed herein is primarily designed to receive the sealed tube 100 pre-comprising the sample collector stick 200 in the second chamber 323, it is understood by a person skilled in the art that the tube 100 devoid of the sample collector stick 200 could be initially inserted into the second chamber 323, and only after that could the sample collector stick 200 be inserted into the tube 100.

[0112] In some embodiments, at least one controller 330 comprises instructions to first activate the mixing mechanism 340 for a first predetermined time. In some embodiments, the first predetermined time is between 1 and 15 minutes, between 1 and 10 minutes, or between 1 and 5 minutes. Each possibility represents a separate embodiment of the present invention. In some embodiments, the predetermined time is programmed according to a relevant protocol. In some embodiments, the predetermined time is when the sample is fully dissolved within the buffer. In some embodiments, the system 300 further comprises a sensor (e.g., an optical sensor) configured to evaluate the dissolution of the sample within the buffer.

[0113] Reference is now made for Figures 7A-7B and Figure 8. In some embodiments, the mixing mechanism 340 comprises vibrations by a vibrator, coupled to the second chamber 323. As exemplified in Figure 7A, the mixing mechanism 340, e.g., vibrator, may be cylindershaped. The vibrator may be positioned horizontally to the second cylinder chamber 323 or vertically, i.e., in a parallel axis to second chamber 323, attached to the bottom, median or upper portion of second chamber 323. It was found herein that optimal and efficient mixing, resulting in the dilution of all of the biological sample within the fluid, occurs when the vibrator is positioned vertically and coupled to the bottom portion of the second cylinder chamber 323.

[0114] As demonstrated in Figure 4 and Figure 5A, the system 300 may further include a flexible ring surface 321 located on or adjacent to the upper side 324 of the container housing 320. This flexible ring surface 321 is designed to encircle the second chamber 323 and / or the collector gripping segment 230, allowing for vibrations of the tube when the mixing mechanism is in operation. The flexible surface 321 is typically composed of soft, sponge-like materials to facilitate tube vibrations. Non-limiting examples for such sponge-like materials are silicone foam, polyurethane foam, neoprene sponge, ethylene-vinyl acetate foam foam rubber, and poron foam.

[0115] After mixing for the first predetermined or preprogrammed time has accomplished, the controller 330 further instructs to automatically activate the valve actuator 351 to open the valve 160 and to dispense a predetermined volume of the homogenous mixture onto the test cassette 380, or more specifically, onto the sample loading area 382 of the test cassette 380.

[0116] The transferring of the valve 160 from a close configuration 161 to an open configuration 162, can be performed by the valve actuator 351 directly, or as exemplified in Figure 7B, by a valve connector 350, attached or connected to the valve actuator 351.

[0117] Non limiting examples for valve actuators include: a) rotational valve actuator consisting of a motor-driven gear system that rotates the valve connector, e.g., electric motors, stepper motors, or servo motors; b) piston valve actuator that uses the linear motion of a piston to control the valve; c) solenoid valve actuator that use electromagnetic force to operate the valve; d) pneumatic or hydraulic valve actuator that use air or liquid pressure to operate the valve. The pressure may be applied to a piston or diaphragm, causing movement that actuates the valve; and, e) electrically controlled valve actuator that use electrical signals to control the valve position. These may incorporate motors, solenoids, or other electromechanical components for precise valve control.

[0118] In some embodiments, the valve actuator 351 comprises a mechanical interface mean that enables engagement to the valve connector 350 and a rotational movement of the valve connector 350 to switch the valve 160 from the closed position 161 to the open position 162.

[0119] To ensure accurate results, it is essential not only to achieve homogeneous dissolution of the biological sample within the buffer and accurately dispense a predetermined volume of the homogeneous solution, but also to ensure that all of the dispensed predetermined volume is absorbed by the sample loading area 382 of the cassette. Therefore, in some embodiments, second chamber 323 comprising the tube 100 is positioned at a first predetermined distanceabove the first chamber 325 comprising the cassette 380, e.g., as illustrated in Figure 8, to allow optimal liquid dripping and accurate absorption of the predetermined volume by the sample loading area 382.

[0120] In some embodiments, after the diluted biologic sample is loaded on the sample loading area 382 it flows along the test strip through capillary action to the test results window 381. For example, if the target analyte is present in the sample, it binds to labeled antibodies or antigens in the conjugate pad. The analyte then binds to capture reagents at the test line(s), causing a visible color change. The appearance of colored lines at specific locations on the membrane indicates a positive result, while a control line confirms that the test is valid.

[0121] In some embodiments, the intensity of the color in the test line(s) within the test results window 381 is positively correlated to the analyte concentration. In some embodiments, accurate results are achieved only when the intensity of the line(s) is captured after a predetermined time. Therefore, the controller 330 disclosed herein comprises instructions to further automatically operate the illumination means and camera 360 to capture an image of the cassette 380 or the test results window 381 after a second predetermined time. In some embodiments, the second predetermined time is dependent on the specific type of the cassette 380. In some embodiments the predetermined time is selected between 1 and 20 min, 1 and 15 min, 1 and 10 min, 1 and 5 min. Each possibility represents a separate embodiment of the present invention.

[0122] As exemplified in Figures 8 and Figure 9A, the illumination and camera means 360 are located at a second predetermined distance above the test result window 381, allowing capturing the entire test results window 381. In some embodiments, illumination means comprise a light-emitting diode (LED). In other embodiments, the LEDs are arranged in a circular fashion, allowing minimal shadow on the test strip or test results window 381.

[0123] As used herein, the term "illumination" refers to the lighting system used to illuminate the test strip or test results window 381 evenly, consistently and with minimal shadow. The illumination system, in conjunction with the opacity of the container housing 320 to external light, creates the controlled lighting environment necessary for capturing the cassette without the need for light pre-calibration. Proper illumination ensures that the colors on the test strip are accurately and repeatably represented when captured by the camera and help to minimize shadows, reflections, and uneven lighting that can affect color detection.

[0124] The image is captured by a camera or by an imaging system. The camera may capture images in RGB (Red, Green, Blue) or other color spaces for color analysis. The camera 360, the controller 330, or both, may comprise an image processing system, for at least partially analyzing the captured image. In yet another embodiment, at least part of the analysis of the image may be performed on a cloud-based system.

[0125] In some embodiments, communication means 370 (e.g., as illustrated in Figure 4), are operably linked to the controller 330 for at least one member of the group selected from: communicating the image and / or image based-data to a cloud-based system configured to analyze the image; and, communicating the image and / or analysis of the image to a remote operator such as the user's smartphone, a medical data system, or analogue system thereof. The communication means 370 may be wireless or non-wireless, although wireless communication means are preferable.

[0126] As used herein, the term "image based-data" encompasses any data or analysis derived from the image. This refers to a wide range of information obtained through image processing, such as measurements, features, patterns, and statistics extracted from the image.

[0127] The term "medical data system" used herein refers to a specialized information management system designed to collect, store, organize, and analyze various types of medical and healthcare-related data. This may include patient records, diagnostic test results, treatment plans, and medication histories. As used herein, the terms "analogue system thereof' and "analogue device thereof refer to a corresponding system / device being capable of at least one of storing, organizing, analyzing, exhibiting, and any combination thereof, of medical-related data.

[0128] In some embodiments, the controller 330 is operated by a remote operator or controller, such as the user's smartphone, or analogue device thereof. In some embodiments, the system 300 further comprises a sensor operably linked to the controller 330. A sensor could be used to detect the presence, the orientation, the type, or any combination thereof, of the test cassette 380 in the first chamber 325. In a yet another embodiment, a sensor could be used to detect the presence, orientation, or both, of the tube 100, within the second chamber 323. In yet another embodiment, a sensor could be used to detect the presence of the sample collector stick 200 within the tube 100. In some embodiments, a sensor could be used to detect the presence of a sample upon the sample collecting segment 220 and / or its dissolution within the buffer. In some embodiments the controller is operably linked to a user display 327, present on thecontainer housing 320, as exemplified in Figure 5B. In some embodiments, the user would be notified by displaying an alert on the user display 327, on the smartphone, or on an analogue device, when any of the parameters selected from the presence, orientation, type, or any combination thereof, of the test cassette 380; the presence, orientation, or both, of the tube 100; the presence of the sample collector stick 200 within the tube 100; or any combination thereof, is not valid.

[0129] Examples for sensors configured to detect the presence, orientation and / or type of an object are known in the art. Presence sensors, detecting the presence or absence of an object within a certain range, include inductive proximity sensors, capacitive proximity sensors, ultrasonic sensors and optical sensors such as photodiodes, phototransistors, and infrared sensors. Orientation sensors detecting orientation of an object, include accelerometers and gyroscopes. Type and / or batch sensors include barcode scanners and RFID (radio-frequency identification) reader. Additionally, the system disclosed herein may utilize combination sensors such as smart sensor systems or integrated sensor systems and internet of things (loT) sensors.

[0130] In some embodiments, a calibration curve is saved in the cloud and used for analysis of each cassette or strip of the same type and batch, for maintaining consistency and accuracy in measurement.

[0131] In some embodiments, the tube 100, the system 300, and both, disclosed herein, are configured for detection of an analyte within a biologic sample. The analyte may be a protein or nucleic acid molecule whose levels are dysregulated, such as for example in a disease. The disease may be an inflammatory disease, an autoimmune disease, or cancer. The inflammation may be systemic inflammation. The analyte may be an analyte used to diagnose a disease or an analyte used to monitor disease progression. In some embodiments, the disease comprises a gastrointestinal disease selected from gastrointestinal cancer, gastrointestinal infection and inflammatory bowel disease (IBD).

[0132] Although the scope of the present invention is mainly addressed to an automatic system configured to detect a test cassette 380, selected from calprotectin test, fecal occult blood test (FOBT), fecal immunochemical test (FIT), stool bacterial culture, Clostridium difficile (C. difficile) toxin test, c-reactive protein (CRP), rotavirus and / or norovirus antigen tests, and parasite tests, and Helicobacter pylori (H. pylori), a skilled in the art would appreciate that anytest cassette or strip, and specifically a test that is based on colorimetric detection, would be compatible to be used by the system disclosed herein.

[0133] Reference is now made to Figures 12A-12E. In some embodiments, the stool sample may be collected by manually contacting the collecting segment 220 of the sample collector stick 200 with the stool sample. In other embodiments, the stool sample may be collected onto the collecting segment 220 using a stool sampling device 400 designed to accommodate the sample collector stick 200. The sample collecting segment 220 may comprise notches or grooves. In some embodiments, the stool sampling device 400 comprises a proximal wiping base 420 and a distal covering portion 411 (Figures 12A-12C), or element as exemplified by feature 412 (Figure 12D), and feature 413 (Figure 12E), configured to allow sampling of a predetermined amount of stool. In some embodiments, the inner diameter of the proximal wiping base 420 is configured to tightly fit the external diameter of the longitudinal segment of the sample collector stick 210, allowing wiping or removal of excessive stool that remains on the sample collector stick 200 when the stick is pulled out of the sampling device 400.

[0134] Reference is still made to Figures 12A-12C. The stool sampling device 400 may comprise a proximal wiping base 420, a longitudinal segment 410 and a covering portion 411 comprising an opening 401 on the lateral side of the distal end of the longitudinal segment 410. During sampling, the stool sample 500 contacts the collecting segment 220 on the sample collector stick 200 only within the opening 401, whereas excessive stool remains on the outer surface of the covering portion 411, allowing stool to be attached only to the sample collecting segment 220, as exemplified in feature 501 of Figure 12C.

[0135] Reference is still made to Figure 12D. In some embodiments, the stool sampling device 400 comprises a proximal wiping base 420 and a covering element 412, e.g., a plug, that is temporally attached to the distal end of the sample collector stick 200, blocking direct contact between the collecting segment 220 and the stool in the distal end of the sample collector stick 200. In some embodiments, the shape of covering element 412 is an inverted cone, wherein the maximal diameter of the cone is in its distal outer portion facing the stool sample. In some embodiments, the maximal diameter of covering element 412 is higher compared to the inner diameter of the proximal wiping base 420, and once the sample collector stick 200 is pulled out of the stool sampling device 400, the covering portion 412 is detached from the stick 200 and may be left within the opening of the proximal wiping base 420. Excessive stool present on the outer surface of the stick 200 remains attached to the inner portion of the proximal wiping base 412, allowing sampling of a predetermined amount of stool. Reference is still madeto Figure 12E. The distal covering element may also be a detachable hollow cylinder shaped 413 temporarily wrapping the distal portion of the sample collector stick 200. The covering element 413 may comprise an opening 402 configured to allow direct contact between the collecting segment 220 and the stool sample. Similarly to covering element 412, covering element 413 may also be detached from the stick 200 once the sample collector stick 200 is pulled out of the stool sampling device 400. Covering portion or element, e.g., as exemplified by features 411, 412, 413, may be made of a flexible material. Non-limiting examples for such a flexible material include silicone elastomers, polyurethane elastomers, thermoplastic elastomers (TPE), natural rubber, fluorosilicone rubber, ethylene propylene diene monomer (EPDM) rubber, etc.

[0136] As used herein, the terms "covering element" and "covering portion" encompass an element and a portion thereof, respectively, configured to block direct contact between the biologic sample and at least one portion of the sample collecting stick 200.

[0137] In some embodiments, collecting the stool sample comprises a manual sampling by contacting the stool sampling device 400 with at least one portion of the stool. In other embodiments, collecting the stool sample is by a stool collecting device such as disclosed in PCT / IL2024 / 051020, taking priority from USP application number 63544210, "Stool Sample Collecting Device and Method Thereof', herein incorporated by reference in its entity.

[0138] In some embodiments, the stool sampling device 400 is accommodated within a stool collecting device 500 configured to collect a homogenous stool sample comprising a stool receiving container 501; and a lid 502 for sealing the container 501, and the collecting the stool sample comprises rotating the lid 502 to obtain the homogenous stool sample.

[0139] As used herein, the term "about" when combined with a value refers to plus and minus 10% of the reference value. For example, a length of about 1000 nanometers (nm) refers to a length of 1000 nm ± 100 nm.

[0140] It is noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0141] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination.EXAMPLE 1

[0142] Each tube of the invention is filled with a known amount of buffer at atmospheric pressure. The initial pressure within the sealed tube is negative due to the gravitational force acting on the fluid. Two forces act on the fluid within the tube and counteract each other: vacuum and gravity. After insertion of the sample collecting stick into the tube via the upper portion, the air pressure within the tube above the fluid level increases. Once the valve is opened, the fluid is dispensed out of the tube via the valve, and the volume of the air above the fluid level increases until it reaches equilibrium, according to the ideal gas law equation PV=RT (P -pressure, V-volume, R- molar gas constant, T- temperature).

[0143] A sample collecting stick with lengh of 22.44mm, and diameter of 4mm, was examined in various temperatures, to examine the effect of viscocity changes. The volume of pressured air within the tube was reduced by 281.49 mm3(or 0.28149 ml) due to the insertion of the sample collecting stick.

[0144] For each test, after the sample collecting stick was fully positioned within the tube, the conical valve was opened.

[0145] Table 1. Volume of dispensed fluid in different examined temperatures

[0146] It is noted that that the typical sample fluid volume that is required for lateral flow tests is between 50pl and 150pl .EXAMPLE 2

[0147] The inventors examined various vibration protocols to determine the optimal operation conditions for extracting stool from the groove(s) in the sample-collecting segment 220 of the sample collection stick 200, following stool collection. The evaluation was conducted using the sample collection stick 200, the tube 100, and the automatic system 300, as disclosed herein. Stool samples with varying textures were used, ranging from Bristol Stool Chart score 1 (constipation) to score 7 (diarrhea). The tube 100 was filled with a buffer compatible with calprotectin testing. The procedure included the following steps: 1) Determine and record the Bristol score; 2) Collect a stool sample using the sample collection stick 200; 3) Evaluate the groove(s) after stool collection; 4) Fully insert the sample collection stick 200 into the tube 100 described herein; 5) Place the tube, containing the sample collection stick, into the automatic system 300 and activate vibration, clockwise (CW) or counter-clockwise (CCW), according to the selected vibration protocol; 6) Remove the stick from the tube and assess whether the stool has been completely removed from the grooves; 7) If stool residues remain in the grooves, repeat steps 4-6, increasing the vibration time by 1 minute with each subsequent attempt, until no residue remains. In the exemplified test, small floating elements composed of polypropylene with a density of about 0.9 g / mL were also evaluated in some of the tests.

[0148] Table 2. Total vibration time needed for sample’s removal from the groove(s) with and without floating elements in the tube.As shown in Table 2, the protocol involving a 15-second CW run followed by a 15-second CCW run was found to be effective, with 5 out of 8 tests resulting in complete sample removal within 4 minutes or less. Moreover, the inventors found that the addition of small floating elements within the tube may help reduce the time required for full extraction of the sample from the grooves.

Claims

CLAIMSWhat we claim is:

1. A tube 100 for receiving a biologic sample, and dispensing a predetermined volume of diluted said biologic sample, comprising: a tube housing 110 configured to accommodate fluid, said tube housing 110 having an upper portion 130 and a lower portion 140; said upper portion 130 comprises a cone 111 comprising an upper opening 116 and a hole 113 at the lower end of said cone 111, said hole 113 is provided with a temporally central sealing element 112; said upper portion 130 is configured for receiving a sample collector stick 200 comprising a longitudinal segment 210 with collector gripping segment 230 and a lower sample collecting segment 220 configured to accommodate a biologic sample; the opening of said tube lower portion 140 comprises a dispensing valve 160, configured in a closed position 161 preventing said fluid from exiting from said opening of said lower portion 140, and operable in an open position 162 to permit said fluid to exit; wherein, once said collector stick 200 is inserted through said hole 113 to a predetermined position, the dimensions of said cone 111 and said hole 113 are configured to create a seal with said longitudinal segment 210 of said collector stick 200, generating a predetermined increased pressure (AP) of air above the fluid, further wherein, said longitudinal segment 210 in said predetermined position comprises an internal segment 211 and an external segment 212, and the dimensions of said internal segment 211 are proportional to the predetermined volume of said fluid exiting from said lower portion 140, for generating dispensing of a predetermined volume of the fluid once said valve 160 is in said open position 162.

2. The tube of claim 1, wherein said predetermined position of said collector stick 200 within said tube is when said lower sample collecting segment 220 extends into said fluid, said gripping segment 230 closes said upper opening 131, or both.

3. The tube of claim 1 or 2, further comprising a ring 132 in said upper opening 131, and said collector gripping segment 230 comprises a sample collector locking element 240, configured to lock said ring 132, when said collector stick 200 is in said predetermined position.

4. The tube of any one of claims 1 to 3, wherein said cone 111 is a press-fitted cone.

5. The tube of any one of claims 1 to 4, further comprising an elastomeric ring 115 at said lower end of said cone 111, and said hole 113 is a cavity within said elastomeric ring 115.

6. The tube of any one of claims 1 to 5, wherein said central sealing element 112 is a silicone plug or an elastomeric analogue thereof.

7. The tube of any one of claims 1 to 6, wherein central sealing element 112 is pressed into said fluid upon insertion of said sample collector stick 200 through said hole 113.

8. The tube of any one of claims 1 to 7, further comprising a mixing mechanism 340, configured to homogenously dilute said biologic sample loaded on said collecting segment 220, within said fluid, and said dispensing is of a predetermined volume of a homogenous mixture of said fluid and said sample.

9. The tube of claim 8, wherein said mixing mechanism is vibrations by an external automatic vibrator.

10. The tube of claim 9, further comprising at least one solid element within said fluid, configured to vibrate when said external automatic vibrator is operated, to facilitate and / or shorten the time required to homogeneously dilute said biological sample loaded onto said collecting segment 220 in said fluid.

11. The tube of claim 10, wherein said at least one solid element has a density lower than 1 gr / ml.

12. The tube of any one of claims 1 to 11, further comprising a lateral external longitudinal projection 150 configured to: a. restrict rotational movement of said tube within a cylinder chamber 323 of an automatic system 300 for preparing and analyzing said biologic sample, said automatic system 300 comprises a valve actuator 351; and, b. place said valve 160 in adjacent to said valve actuator 351.

13. The tube of any one of claims 1 to 12, wherein the ratio (v / v) between the volume of said internal segment 211 of said sample collector stick 200, and said predetermined volume of said fluid is between 5: 1 to 1:5 in room temperature.

14. The tube of claim 13, wherein said ratio is about 1: 1.

15. The tube of any one of claims 1 to 14, wherein said biologic sample is a stool sample.

16. An automatic system 300 for preparing diluted biologic sample and detecting an analyte in said biologic sample, comprising: a container housing 320; a first chamber 325 configured for receiving a test cassette 380 of a biologic analyte; a second chamber 323 configured for receiving a tube 100, said tube 100 is configured for receiving a sample collector stick 200 comprising a biologic sample, said tube 100 comprises an upper portion 130 comprising a cone 111 comprising an upper opening and a hole113 at the lower end of said cone 111, said hole 113 is provided with a temporally central sealing element 112, the opening of said tube lower portion 140 comprises a dispensing valve 160, configured in a closed position 161 preventing said fluid from exiting from said opening of said lower portion 140, and operable in an open position 162 to permit said fluid to exit; at least one controller 330, operably linked to: a. a mixing mechanism 340 configured to homogenously dilute said biologic sample within said fluid 120; b. a valve actuator 351 configured to open said valve 160 after said mixing and to dispense a predetermined volume of homogenous mixture; c. illumination means and camera 360 configured to capture an image of said test cassette 380; and, d. optionally user display 327 on said container housing 320, wherein said at least one controller 330 comprises instructions to activate said mixing mechanism 340 for a first predetermined time, to further automatically activate said valve actuator 351 to open said valve 160 and to dispense a predetermined volume of said homogenous mixture onto said test cassette 380, and to further automatically operate said illumination means and camera to capture an image of said test cassette 380 after a second predetermined time, to produce an automatic preparation and detection of said biologic sample.

17. The automatic system of claim 16, wherein said test cassette 380 is configured to produce color development proportional to concentration of said biologic analyte in said biologic sample, and said illumination means and camera 360 are configured to capture an image of said color development within said test cassette 380.

18. The automatic system of claim 16 or 17, wherein said at least one controller 330 is operably linked to a remote operator via communication means 370, said remote operator is configured to perform at least one member of the group consisting of: a. analyzing said image and / or image based-data; b. storing at least one member of the group consisting of said image, said image based-data and analysis of said image based-data; and, c. displaying at least one member of the group consisting of said image, said image based-data and analysis of said image based-data in at least one of a user's smartphone, a medical data system, or analogue system thereof.

19. The automatic system of claim 18, wherein said remote operator is a cloud-based system further comprising data of a standard curve of said analyte, for a specific batch of said test cassette, and analyzing of said image and / or image based-data is based on said standard curve.

20. The automatic system of claim 18, wherein said at least one controller 330 is operated by said remote operator such as the user's smartphone, or analogue device thereof.

21. The automatic system of any one of claims 16 to 20, wherein said test cassette 380 is a lateral flow cassette.

22. The automatic system of claim 21, wherein said lateral flow cassette comprises a sample loading area 382 and a test results window 381.

23. The automatic system of claim 22, wherein said second chamber 323 is positioned at a first predetermined distance above said first chamber 325, to allow optimal liquid dripping and accurate absorption of said predetermined volume by said sample loading area 382, once said tube 100 is in said second chamber 323.

24. The automatic system of claims 22 or 23, wherein said illumination and camera means 360 are located at a second predetermined distance above said test result window 381.

25. The automatic system of any one of claims 16 to 24, further comprising at least one sensor operably linked to said at least one controller 330, said at least one sensor detects at least one member of the group consisting of: the presence, the orientation, the type, or any combination thereof, of said test cassette 380, the presence, orientation, or both, of said tube 100, the presence of said sample collector stick 200 within said tube 100, the presence and / or dissolution of said sample within said buffer 120, and any combination thereof.

26. The automatic system of claim 25, further comprising a sensor configured to detect the dissolution of the sample within said buffer 120, and said first predetermined time is when said dissolution of the sample within said buffer is above a predetermined threshold, creating a homogenous solution.

27. The automatic system of any one of claims 16 to 26, further comprising a lid 310, wherein closure of said lid 310 enables at least one member of the group consisting of: noise reduction, minimal user contact, a complete sealing of said system from external light, or any combination thereof.

28. The automatic system of any one of claims 16 to 27, wherein said valve actuator 351 is connected to a valve connector 350.

29. The automatic system of claim 28, wherein said valve actuator 351 comprises a mechanical interface mean that enables engagement to said valve connector 350 and a rotational movement of said valve connector 350 to switch said valve 160 from said closed position 161 to said open position 162.

30. The automatic system of claim 28 or 29, further comprising a longitudinal notch 322, compatible to an external longitudinal projection 150 positioned alongside said tube 100, saidnotch 322 is configured to mark insertion site for a user, by restricting rotational orientation of said tube 100, allowing position of said valve connector 350 adjacent to said valve 160.

31. The automatic system of any one of claims 16 to30, further comprising a flexible ring surface 321, on or adjacent to the upper side 324 of said container housing 320, configured to encircle said tube 100 and / or the collector gripping segment 230 and to allow vibrations of said tube 100, once said mixing mechanism operates.

32. The automatic system of any one of claims 16 to 31, wherein said mixing mechanism 340 comprises vibrations by a vibrator, coupled to said second chamber 323.

33. The automatic system of claim 32, wherein said vibrator is positioned vertically and coupled to a bottom portion of said second chamber 323.

34. The automatic system of any one of claims 16 to 33, wherein said biologic sample is a stool sample.

35. The automatic system of any one of claims 16 to 34, wherein said biologic analyte is a protein or nucleic acid whose levels are dysregulated in a gastrointestinal disease selected from the group consisting of: gastrointestinal cancer, gastrointestinal infection and inflammatory bowel disease (IBD).

36. The automatic system of any one of claims 16 to 35, wherein said test cassette 380 is selected from the group consisting of calprotectin test, fecal occult blood test (FOBT), fecal immunochemical test (FIT), stool bacterial culture, Clostridium difficile (C. difficile) toxin test, c-reactive protein (CRP), rotavirus and / or norovirus antigen tests, parasite tests, Helicobacter pylori (H. pylori) and any combination thereof.

37. A method for preparing diluted biologic sample and detecting an analyte in said biologic sample, the method comprising: a. obtaining a sample collector stick 200 comprising a biologic sample within a sample collecting segment 220; b. obtaining a tube 100 comprising: fluid 120; an upper portion 130; and a lower portion 140, said upper portion 130 comprises an upper opening 131 and a hole 113 provided with a temporally central sealing element 112; the opening of said tube lower portion 140 comprises a dispensing valve 160, configured in a closed position 161 preventing said fluid from exiting from said opening of said lower portion 140, and operable in an open position 162 to permit said fluid to exit; c. inserting said sample collector stick 200 into said tube 100 to a predetermined position via said hole 113; d. obtaining an automatic system 300 comprising:a first chamber 325 configured for receiving a test cassette 380 of a biologic analyte, said test cassette 380 is configured to produce color development proportional to concentration of said biologic; a second chamber 323 configured for receiving said tube 100; at least one controller 330, operably linked to: i. a mixing mechanism 340 configured to prepare a homogenous mixture of said biologic sample and said fluid 20, within said tube 100; ii. a valve actuator 351 configured to open said valve 160 after said mixing and to dispense a predetermined volume of said homogenous mixture; iii. illumination means and camera 360 configured to capture said color development within said test cassette; and iv. optionally user display 327 on said container housing 320, wherein said at least one controller 330 comprises instructions to activate said mixing mechanism 340 for a first predetermined time, to further automatically activate said valve actuator 351 to open said valve 160 and to dispense a predetermined volume of said homogenous mixture onto said test cassette 380, and to further automatically operate said illumination means and camera to capture said test cassette 380 after a second predetermined time, to produce an automatic preparation and analysis of said biologic sample; e. turning on said automatic system 300; f. inserting a lateral flow cassette 380 into said test cassette chamber 325, inserting said tube 100 comprising said sample collector stick 200 into said second chamber 323; and, g. obtaining at least one member of the group consisting of: an image, an image-based data, and analysis of said image-based data, in at least one of user's smartphone, a medical data system, or a corresponding system thereof.

38. The method of claim 37, wherein said step (f) comprises fully inserting said tube 100 comprising said sample collector stick 200 into said second chamber 323, allowing said valve actuator 351 to be positioned adjacent to said valve 160.

39. The method of claim 37 or 38 further comprising at least one sensor operably linked to said at least one controller 330, said at least one sensor detects at least one member of the group consisting of: the presence, the orientation, the type, or any combination thereof, of said test cassette 380, the presence, orientation, or both, of said tube 100, the presence of said sample collector stick 200 within said tube 100, the presence and / or dissolution of the sample within said buffer 120, and any combination thereof.

40. The method of any one of claims 37 to 39, wherein said user's smartphone or analogue device thereof displays at least one notification that validates said at least one member of the group consisting of: the presence, the orientation, the type, or any combination thereof, of said test cassette 380, the presence, orientation, or both, of said tube 100, the presence of said sample collector stick 200 within said tube 100, the presence and / or dissolution of the sample within said buffer 120, and any combination thereof.

41. The method of any one of claims 37 to 40, wherein said biologic sample is a stool sample and said step (a) further comprises collecting said stool sample on said sample collecting segment 220 by a stool sampling device 400 configured to accommodate said sample collector stick 200, said stool sampling device 400 comprises a proximal wiping base 420 and a distal covering element or portion 411, 412, or 413, configured to sample a predetermined amount of said stool on said collecting segment 220.

42. The method of claim 41, wherein said collecting said stool sample comprises a manual sampling by contacting said stool sampling device 400 with at least one portion of said stool.

43. The method of claim 41, wherein said stool sampling device 400 is accommodated within a stool collecting device 500 configured to collect a homogenous stool sample comprising a stool receiving container 501; and a lid 502 for sealing said container 501, and said collecting said stool sample comprises rotating said lid 502 to obtain said homogenous stool sample.

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