Tissue Sampling Kit and Device

Abstract

TISSUE SAMPLING KIT. The present invention relates to a sampling kit comprising a reaction chamber (20) and a sampling device (1) configured to be mounted in the reaction chamber and to provide a defined reaction volume (16) between the base of the reaction chamber and the sampling device. The sampling device has a proximal handling part (3) and a distal scraping part (4) comprising a scraping formation (9) configured to collect a tissue sample when the distal scraping part is rubbed against a tissue surface. The sampling device comprises an internal lumen configured to supply reaction fluid from a distal end of the tissue sampling device to the reaction volume through a distal opening (7) disposed in the distal scraping part when the tissue sampling device is mounted in the reaction chamber.An automated high-throughput sampling and indexing system is also described.

Description

1 / 28 Tissue Sampling Kit and Device Field of Invention

[001] The present invention relates to a sampling kit for use with tissues, such as meat carcasses used in the food industry. The invention also relates to a method for tissue sampling. Background of the Invention

[002] Tissue sampling kits are used in the meat industry to take a tissue sample from a carcass, which can then be used to extract carcass-specific DNA. This DNA can be used to determine specific carcass information, such as the origin of the meat (i.e., country, herd, farm, slaughterhouse), or to determine the source of a microbial pathogen. The uses of tissue sampling kits, and a specific tissue sampling kit, are described in WO 2008 / 034847 (Parlanca Limited). The kit comprises a tissue sampling device and a lid to receive the device after a sampling operation. The sampling device has a handle part and a sampling part with a scraping edge for scraping tissue from a carcass. The scraping part is provided by a series of deep circumferential edges that tear the carcass tissue (as opposed to scraping only a few meat fibers).Following a tissue sampling operation, the cap is placed on the device to protect the tissue sampling portion from contamination during transport to a laboratory. In the laboratory, the cap is removed, and the tissue sampling portion is placed in a well of a multi-well plate that already contains reaction fluid. The device is designed to nest snugly and securely in the well. In the well, the reaction fluid reacts with the tissue to perform cell cytolysis and release DNA during an incubation period. After the... Petition 870250090762, dated 06 / 10 / 2025, page 21 / 99 During the 2 / 28 incubation period, the device is removed from the well, and a sample of the liquid in the well, containing the cell-free DNA, is removed for analysis. During this process, retraction of the device from the well may result in droplets of the removed liquid falling onto the plate and contaminating other wells on the plate, resulting in well contamination and corrupted analysis results.

[003] US 4707450 describes a specimen collection and testing unit provided for use in collecting a biological specimen. The specimen collection and testing unit comprises an elongated swab having a fibrous tip mounted at one end of an elongated hollow shaft. The opposite end of the swab shaft is borne by a hollow base, usually cylindrical and enlarged, formed from a plastic or similar material, and having sufficient resilience to permit manual compression and bending without exhibiting significant plastic deformation. The cylindrical base contains one or more reagents preserved within one or more sealed frangible ampoules.

[004] During use, the swab and base are manually handled as a unit for collecting a biological specimen, or similar, on the fibrous tip of the swab, without removing the swab from the cylindrical base. After the specimen is collected, the cylindrical base is manually bent, or otherwise deformed, to break the frangible ampoule(s), thus releasing the reagents contained therein, which can be properly mixed by gently shaking the base. The reagents will then be available for pumping through the hollow stem of the swab to the fibrous tip by applying manual pressure to the cylindrical base.

[005] Once incubated, the test unit needs to be removed from the reagent chamber to allow the fluid contained in the chamber to be sampled, thus exposing the chamber and the system to cross-contamination. Petition 870250090762, dated 06 / 10 / 2025, page 22 / 99 3 / 28

[005] The pasteurization temperature must be maintained, depending on the product to be pasteurized, in a range substantially between 57 °C and 72 °C to avoid product degradation at higher temperatures or to avoid insufficient pasteurization when using lower temperatures.

[006] It is an objective of the invention to overcome at least one of the problems mentioned above. Summary of the Invention

[007] The applicant has addressed the problems of the prior art by providing a sampling device or kit comprising a sampling device and a reaction chamber (e.g., a well or a lid), wherein the sampling device has a hollow lumen that allows fluid to be added to, or removed from, the reaction chamber using one or more openings, while the tissue sampling device remains nested within the reaction chamber. The hollow lumen generally extends from an upper (proximal) opening to one or more openings disposed in the sampling (distal) portion of the device. During use, the sampling device containing the scraped tissue can be nested snugly within the well of a multi-well plate (or in an individual well provided by a lid), and the reaction fluid then added to the well through the lumen of the sampling device.In this way, the reaction liquid can only be added after the sampling device is mounted in the reaction chamber, thus limiting the risk of spillage. It is important to note that providing a sampling device with an internal lumen allows the liquid to be removed from the well through the lumen without needing to remove the sampling device from the well, thus addressing the contamination problems associated with prior art devices. The kit of the invention can take two forms... Petition 870250090762, dated 06 / 10 / 2025, page 23 / 99 4 / 28 one in which the reaction chamber is provided by a lid within which a sampling device is mounted (figures 4A to 6). The lid can be configured to perform detachable mounting on the distal scraping part of the device (to act as a reaction chamber), but also to perform detachable mounting on the proximal handling part of the device, to act as a handle extension in order to facilitate use during a sampling operation. The invention also provides, as described below, an automated tissue sampling system comprising a robotic arm.

[008] In a first aspect, the invention provides a sampling kit comprising a reaction chamber and a sampling device configured to be mounted in the reaction chamber and to provide a defined reaction volume between the base of the reaction chamber and the sampling device. The sampling device has a proximal handling part and a distal scraping part, which comprises a scraping formation configured to collect a tissue sample when the distal scraping part is rubbed against a tissue surface. The kit is characterized in that the sampling device generally comprises an internal lumen configured to supply the reaction fluid from a distal end of the sampling device to the reaction volume through a distal opening (e.g., a window or partition) disposed in the distal scraping part when the sampling device is mounted in the reaction chamber.

[009] Such a tissue sample is, for example, made up of meat tissue fibers. The scraping formation, according to the description, is therefore suitable for collecting meat tissue fibers.

[010] In one embodiment, the reaction chamber comprises a lid for the tissue sampling device. Petition 870250090762, dated 06 / 10 / 2025, page 24 / 99 5 / 28

[011] In one embodiment, the cap is configured to be detachably attached to the proximal handling end of the tissue sampling device in order to provide a handling extension.

[012] In one embodiment, an external base of the cover includes an electronic identification indicator that can be read by a flat scanner. The indicator can be, for example, an RFID indicator or a barcode.

[013] In another embodiment, the reaction chamber comprises a well of a multi-well plate.

[014] In one embodiment, the sampling device is substantially hollow and provides an internal lumen that extends from an open top to at least one opening in a side wall of the distal sampling portion.

[015] In one embodiment, at least two openings are provided in the side wall of the distal sampling portion. The at least two openings may be arranged on opposite sides of the device.

[016] The inner lumen is typically configured to receive a pipette tip, syringe or needle, or any suitable device that can be used to remove liquid from the reaction volume, or introduce liquid into the reaction volume. This allows a pipette tip or any suitable device to be inserted into the lumen and extend through the lumen into the reaction fluid within the lumen. In this way, it is possible to add liquid to the reaction chamber, and / or remove liquid from the reaction chamber, through the lumen, thus allowing the digestion / solubilization of the tissue without removing the sampling device from the well / lid. In other words, the inner lumen is configured to supply the reaction liquid and to subsequently remove the reaction liquid mixed with the tissue sample (reaction mixture or product produced by the reaction mixture) when the device is removed. Petition 870250090762, dated 06 / 10 / 2025, page 25 / 99 6 / 28 Tissue sampling is mounted in the reaction chamber (well or lid, where the lid can be arranged in a well of a multi-well plate, for example).

[017] In one embodiment, the distal opening, in the distal sampling portion, is arranged proximal to the scraping formation.

[018] In one embodiment, the distal scraping part comprises a conical end part.

[019] In one embodiment, the scraping formations comprise one or more wholly or partially circumferential grooves. In one embodiment, the scraping formation has a depth of not more than 1 mm.

[020] In one embodiment, one or more totally or partially circumferential grooves are arranged on the conical end of the distal scraping part. In one embodiment, the circumferential grooves have a depth of not more than 1 mm, 0.9 mm, 0.8 mm, 0.7 mm, 0.6 mm, or 0.5 mm. In one embodiment, the circumferential grooves have a depth of about 0.2 to 1.0 mm. In one embodiment, the circumferential grooves have a depth of about 0.3 to 0.7 mm, or 0.3 to 0.6 mm.

[021] In one embodiment, one or each of the distal openings is disposed on the conical end portion of the distal scraping part, typically proximal to the scraping formations.

[022] In one embodiment, the sampling device and the reaction chamber are configured to ensure engagement. In one embodiment, the sampling device and the reaction chamber are configured to achieve sealed engagement, typically by means of a friction seal fitting. In one embodiment, the sampling device and the reaction chamber are configured to be mounted by means of a press-fit fitting.

[023] In one embodiment, the sampling kit includes a plurality of sampling devices. In one embodiment, each of the plurality Petition 870250090762, dated 06 / 10 / 2025, page 26 / 99 7 / 28 of the sampling devices are detachably mounted in a well of a multi-well plate.

[024] In one embodiment, the multi-well plate includes an electronically identifiable indicator. The indicator may be, for example, an RFID indicator or a barcode.

[025] In another aspect, the invention provides a sampling device having a proximal handling part and a distal scraping part, comprising a plurality of scraping formations configured to collect a tissue sample when the distal scraping part is rubbed against a tissue surface, wherein the distal scraping part comprises a conical end part and the scraping formations comprise one or more totally or partially circumferential edges having a depth of not more than 1 mm.

[026] In one embodiment, the circumferential edges are provided by grooves. In another embodiment, the circumferential edges are provided by protrusions.

[027] In one embodiment, the circumferential protrusions have a depth of not more than 1 mm, 0.9 mm, 0.8 mm, 0.7 mm, 0.6 mm or 0.5 mm.

[028] In one embodiment, the circumferential edges have a depth of about 0.3 to 1.0 mm. In one embodiment, the circumferential grooves have a depth of about 0.3 to 0.7 mm, or 0.3 to 0.6 mm.

[029] In another aspect, the invention provides a method for sampling and treating tissue comprising the steps of: provide a sampling kit comprising a reaction chamber and a sampling device configured to be mounted in such a way Petition 870250090762, dated 06 / 10 / 2025, page 27 / 99 8 / 28 detachable to the reaction chamber, and provide a defined reaction volume between the base of the reaction chamber and the sampling device, the sampling device having a proximal handling part and a distal scraping part, which comprises a scraping formation configured to collect a tissue sample when the distal scraping part is rubbed against a tissue surface; To perform a tissue sampling operation by scraping the distal scraping part of the sampling device against the tissue, where a tissue sample is collected by the scraping formation; to detachably mount the tissue sampling device in the reaction chamber, characterized by the fact that the method includes the following steps: Add reaction fluid to the reaction volume through an internal lumen configured to supply the reaction fluid from a distal end of the tissue sampling device to the reaction volume through a distal opening disposed in the distal scraping portion, when the sampling device is mounted in the reaction chamber, where the reaction fluid comes into contact with the tissue sample to provide a reaction mixture in the reaction volume; and incubate the reaction mixture for an appropriate period of time.

[030] Generally, the reaction liquid addition step comprises adding sufficient liquid to at least partially, and usually completely, submerge the distal opening. This means that the liquid in the reaction chamber extends into at least the bottom of the inner lumen, allowing the liquid to be removed using a pipette inserted into the inner lumen. In one embodiment, the reaction liquid formulated to perform cell cytolysis is added to the reaction chamber through the sampling device, where it mixes with the tissue sample that performs the Petition 870250090762, dated 06 / 10 / 2025, page 28 / 99 9 / 28 cytolysis of the cells releases cellular DNA, which diffuses through the fluid, including diffusion into the fluid contained in the inner lumen of the sampling device, and allowing it to be removed through the inner lumen.

[031] In one embodiment, the method includes a step of removing a portion of the reaction mixture or a product produced by the reaction mixture.

[032] In one embodiment, the reaction chamber comprises a lid for the tissue sampling device, wherein the method includes a lid disposition step, with the tissue sampling device mounted on the lid, in a well of a multi-well plate, before adding the reaction liquid to the reaction volume.

[033] In one embodiment, the cap is configured to be removablely attached to the proximal handling end of the tissue sampling device to provide a handling extension, and where the method includes a step of attaching the cap to the proximal handling end of the tissue sampling device before the tissue sampling operation, and releasing the cap from the proximal handling end after the sampling operation.

[034] In one embodiment, the reaction chamber comprises a well of a multi-well plate, and where the method includes a detachable assembly step of the tissue sampling device to the well of the multi-well plate after the sampling operation.

[035] In another aspect, the invention relates to the sampling device only.

[036] In one embodiment, the sampling device has a proximal handling portion and a distal scraping portion comprising a plurality of scraping formations configured to collect a sample. Petition 870250090762, dated 06 / 10 / 2025, page 29 / 99 10 / 28 of the tissue when the distal scraping portion is rubbed against a tissue surface. The distal scraping portion comprises a conical end portion and the scraping formations comprise one or more edges, wholly or partially circumferential, having a depth of not more than 1 mm.

[037] In one embodiment, the circumferential edges are provided by grooves.

[038] In one embodiment, the circumferential edges have a depth of no more than 0.5 mm.

[039] In one embodiment, the circumferential edges have a depth of about 0.3 to 0.7 mm.

[040] In another aspect, the invention provides a system for performing high-throughput indexing sampling of multiple tissue samples, for example, carcasses, comprising: a multi-well plate having a plurality of wells; a plurality of sampling devices nested in the wells of the multi-well plate, each sampling device having a proximal handling part and a distal scraping part nested in the well; a robotic system comprising a robotic arm configured to engage a proximal handling portion of a first tissue sampling device while seated in a first well of a multi-well plate, lift the first tissue sampling device from the first well, move the first tissue sampling device toward a first tissue sample and scrape the distal scraping portion against the first tissue sample, and replace the distal scraping portion of the sampling device in the first well; a sensor for detecting the electronic ID of an electronic identification indicator disposed in the first tissue sample; Petition 870250090762, dated 06 / 10 / 2025, page 30 / 99 11 / 28 a processor operationally coupled to the robotic system and the sensor and configured to receive the detected electronic ID from the sensor and assign the detected electronic ID to the tissue sample with the multi-well plate and the first well identification; and a display system to display the tissue sample ID and associated plate and the first well identification.

[041] Typically, the tissue sample is a carcass, for example, a meat carcass for use in the food industry.

[042] In one embodiment, the multi-well plate and the plurality of tissue sampling devices form part of a tissue sampling kit, according to the invention.

[043] In one embodiment, the system is configured to sample multiple tissue samples (i.e., multiple carcasses) using multiple tissue sampling devices and, optionally, multiple multiwell plates, and to assign a plate and well ID to each tissue sampled.

[044] In one embodiment, the system comprises a liquid-handling robot configured to simultaneously move the reaction fluid into a plurality of wells in a multi-well plate. In another embodiment, the system comprises a liquid-handling robot configured to simultaneously remove a portion of a reaction mixture, or of a product produced by the reaction mixture, from the plurality of wells in the multi-well plate.

[045] Other aspects and preferred embodiments of the invention are defined and described in the other claims set forth below. Brief Description of the Figures

[046] Figure 1 is a perspective view of a device of Petition 870250090762, dated 06 / 10 / 2025, page 31 / 99 12 / 28 sampling, according to the invention. This sampling device uses a well of a multi-well plate as a reaction chamber, and is suitable for use with an automated sampling system (i.e., a sampling robot).

[047] Figure 2 is an end view of the sampling device of Figure 1 facing the handling part of the device.

[048] Figure 3A is a side elevation view of the sampling device of Figure 1 with the internal lumen illustrated by dashed lines, Figure 3B is a cross-sectional view of the sampling device of Figure 1.

[049] Figures 4A, 4B and 5 are elevation views of a sampling kit according to the invention, comprising a sampling device and a detachable cover for the sampling device, which functions as a handle extension in a first configuration (Figures 4A and 4B), and a reaction chamber in a second configuration (Figure 5). This embodiment of the invention is suitable for manual (non-automated) use.

[050] Figures 6A to 6E illustrate a tissue sampling and treatment process of the invention employing the sampling kit of Figures 4A, 4B and 5 (where the reaction chamber is provided by a lid): Figure 6A illustrates the tissue sampling kit from Figure 4 scraping against the surface of a meat carcass to remove tissue samples from the surface; Figure 6B illustrates the cap removed from the distal loop portion of the device and mounted on the proximal tissue scraping portion of the device; Figure 6C illustrates the assembly of Figure 6B nested in a straight position in a well of a multi-well plate; Figure 6D illustrates the reaction fluid being pipetted into the reaction chamber through the inner lumen of the tissue sampling device; Petition 870250090762, dated 06 / 10 / 2025, page 32 / 99 13 / 28 Figure 6E illustrates the liquid being removed from the reaction chamber through the internal lumen in the tissue sampling device.

[051] Figures 7A to 7D illustrate a tissue sampling and treatment process, according to an alternative embodiment of the invention employing the sampling device of Figures 1 to 3B (where a well of a multi-well plate is used as a reaction chamber): Figure 7A illustrates the tissue sampling device from Figure 1 scraping against the surface of a meat carcass to obtain a tissue sample from the surface; Figure 7B illustrates the tissue sampling device of Figure 1 nested in a straight line within a well of a multi-well plate; Figure 7C illustrates the reaction fluid being pipetted into the well of the multi-well plate through the inner lumen of the tissue sampling device; Figure 7D illustrates the liquid being removed from the reaction chamber through the internal lumen in the tissue sampling device.

[052] Figure 8 is a cross-sectional view of a sampling device of figures 1 to 3B nested in a well of a multi-well plate with a defined reaction volume between the well base and the sampling device, and the reaction fluid added to the well to a height H submerging the openings in the device.

[053] Figure 9 is an illustration of a high-throughput, automated carcass tissue sampling and indexing system according to the invention, employing the sampling kit illustrated in Figures 1 to 3B and 8. Detailed Description of the Invention

[054] All publications, patents, patent applications and other Petition 870250090762, dated 06 / 10 / 2025, page 33 / 99 14 / 28 references mentioned in this document are incorporated by reference in their entirety for all purposes as if each individual publication, patent or patent application were specifically and individually indicated as incorporated by reference and the contents thereof mentioned in their entirety. General Definitions and Preferences

[055] Where used and, unless specifically indicated otherwise, the following terms shall have the following meanings, in addition to any broader (or narrower) meaning the terms may have in the art.

[056] Unless otherwise required by the context, the use of the singular herein should be considered as including the plural and vice versa. The term one, an used in relation to an entity should be considered as referring to one or more of that entity. As such, the terms one (or an), one or more and at least one are used interchangeably herein.

[057] As used herein, the term comprise or variations thereof, such as comprises or comprising, shall include any integer mentioned (e.g., an accessory, an element, a feature, a property, a step or method / process limitation) or a group of integers (e.g., accessories, elements, features, properties, steps or method / process limitations), but not the exclusion of any other integer or group of integers. Thus, as used herein, the term comprising is inclusive and has an open meaning and does not exclude additional integers, or method / process steps not mentioned.

[058] As used herein, the term tissue refers to any biological tissue, especially meat used in the food industry, such as carcasses of beef, poultry, lamb, pork, game meat, horse meat or parts thereof. Petition 870250090762, dated 06 / 10 / 2025, page 34 / 99 15 / 28

[059] As used herein, the term reaction fluid refers to a liquid used to interact with the tissue sample and cause a reaction. Generally, the reaction fluid is a reagent for performing cell cytolysis. Such reagents are described in Islam et al (Micromachines March 2017, 8(3):83) Exemplification

[060] The invention will now be described with reference to the specific Examples. These are merely illustrative and serve illustrative purposes only: they should in no way limit the scope of the claimed monopoly or of the invention described. These examples constitute the best currently contemplated way of practicing the invention.

[061] With reference to the drawings, and initially to figures 1 to 3B, a first embodiment of a sampling device is illustrated, which is part of a sampling kit according to the invention, and generally indicated by the numerical reference 1. This embodiment of the sampling device is suitable for use with a well of a multi-well plate as a reaction chamber (as illustrated below in figures 7 and 8) and is suitable for use with an automated sampling system described below. The device comprises an elongated projectile-shaped body 2 having a proximal handle portion 3 and a distal tissue scraping portion 4. The device is hollow and has an internal lumen 5 that provides fluid communication between an open top 6 and one or more openings 7, for example, two openings arranged on opposite side walls of the distal tissue scraping portion 4. The internal lumen is illustrated by dashed lines A in figure 3A.A distal end 8 of the device includes a scraping formation 9 that provides scraping edges configured to collect tissue fibers when the distal scraping portion is scraped against a carcass.

[062] For example, scraping formation 9 comprises a series of Petition 870250090762, dated 06 / 10 / 2025, page 35 / 99 16 / 28 five circumferential grooves spaced along the length of the distal tissue scraping part 4, and located between the end of the device (distal end 8) and the openings 7 of the scraping part 4. A larger number of grooves (e.g., between six and ten grooves) can be used to increase the amount of tissue samples collected.

[063] A smaller number of grooves (for example, between one and four grooves) can be used to reduce the size of the tissue scraping part 4.

[064] In particular, the scraping formation 9 and the openings 7 can be adapted to ensure that some reaction fluid mixed with the tissue sample extends to the bottom of the inner lumen and can be removed through the lumen. The amount of reaction fluid that can be inserted through the inner lumen can also be adapted.

[065] A number of five grooves is, therefore, a good compromise between the amount of tissue samples collected and the size of the tissue scraping part 4.

[066] For example, the circumferential grooves may have a width of about 0.5 mm to about 0.7 mm. The ridge between two grooves may have a width of about 0.5 mm to about 0.7 mm. As an example, the first groove (located near the end of the device) has a width of 0.55 mm, the other grooves have a width of 0.6 mm, the ridge between two grooves has a width of 0.55 mm, and the ridge / rim between the top groove (fifth groove located near opening 7) and the rest of the sampling device has a width of 0.50 mm.

[067] Circumferential grooves can have a depth of about 0.3 mm to about 0.5 mm. These dimensions can be considered effective for removing a sufficient amount of tissue fibers from a meat carcass without tearing the carcass tissue, regardless of Petition 870250090762, dated 06 / 10 / 2025, page 36 / 99 17 / 28 amount of force used during the sampling operation, and without compromising the geometric strength of the sampling device (which is substantially hollow).

[068] In another embodiment, the scraping formation 9 comprises a helical groove or thread.

[069] For example, as illustrated in figure 3B, the inner lumen 5 of the sampling device may have a main section 51 in the shape of a cylinder with, for example, an angle of inclination of 1. The lumen extends from the upper opening 6 to the distal scraping part 8 of the device. The diameter of the cylinder at the upper opening may be about 4 to 6 mm, for example, 4.17 mm. The diameter of the cylinder at the lower end may be about 3 to 4 mm, for example, 3.42 mm.

[070] The tilt angle of the inner lumen can be used to create a frictional fit between the sampling device and a gripping pin of a robotic end-of-arm tool (EOAT), as described below. In this way, the sampling device can be handled even in an unsupervised setting, for example, in a slaughterhouse. In particular, such a tilt angle helps ensure effective robotic operation each time the EOAT engages the sampling device while removing and inserting it into a well of the multi-well plate during the tissue sampling process.

[071] The internal lumen 5 of the sampling device may also have one or more sections in the shape of a truncated cone at the upper opening 6. In the example illustrated in Figure 3B, the internal lumen 5 of the sampling device has a first section 52 in the shape of a first truncated cone and a second section 53 in the shape of a second truncated cone, the smaller diameter of the second truncated cone being the larger diameter of the first. Petition 870250090762, dated 06 / 10 / 2025, page 37 / 99 18 / 28 truncated cone, for example, 4.59 mm, and the smaller diameter of the first truncated cone being the larger diameter of the cylinder, for example, 4.17 mm.

[072] The inner lumen 5 of the sampling device may also have an end section 54 in the distal scraping part 4, which may coincide with the shape of the scraping part 4. For example, the distal scraping part of the device is usually parabolic, and the end section 54 of the inner lumen 5 is a parabolic end section.

[073] As already mentioned, the inner lumen 5 comprises at least one opening 7.

[074] According to a particular embodiment, at least one of said openings 7 is located on a side wall of the distal scraping part. For example, two openings are provided on opposite side walls of the distal scraping part.

[075] In one embodiment, the inner lumen 5 comprises at least one opening located above the scraping formation, not in the scraping formation. For example, such openings may be positioned near the scraping formation, at least 2 mm, and preferably less than 1 mm from the upper groove of the scraping part. In one embodiment, such openings may be in fluid communication with the upper groove of the scraping part. Such location of said at least one opening outside the scraping formation avoids blockage of the openings with tissue samples while collecting a tissue sample by scraping the tissue surface (e.g., the carcass of an animal). According to at least one embodiment, such location also avoids blockage of the pipette / needle when a pipette / needle is used to remove liquid through the lumen.

[076] According to at least one embodiment, such location of said at least one opening close to the scraping formation allows the Petition 870250090762, dated 06 / 10 / 2025, page 38 / 99 19 / 28 openings are completely submerged in the diffused reaction fluid (e.g., the reaction fluid for performing cell cytolysis mixed with sample tissue cells and releasing cellular DNA) of the reaction chamber. In this way, the diffused reaction fluid extends at least into the bottom of the inner lumen, allowing the diffused reaction fluid to be removed using, for example, a pipette inserted into the inner lumen.

[077] In one embodiment, the distal scraping portion of the device is generally parabolic. In this way, at least one opening located in the distal scraping portion is not blocked by a reaction chamber wall when the sampling device is inserted into the reaction chamber.

[078] Various openings can be provided.

[079] A small number of openings (e.g., one or two) is sufficient to insert and remove fluid from the reaction chamber. For example, one opening may be provided at the lower end of the distal scraping part and one or more openings may be provided on a side wall of the distal scraping part. In another example, several openings may be provided on a side wall of the distal scraping part. The distance between the lower end of the distal scraping part and each opening may be different.

[080] A higher number of openings (e.g., three to five) may also be provided, for example, to improve liquid distribution in the reaction chamber.

[081] The openings are designed to be large enough to allow aqueous liquids to drain freely out of the openings, for example, under the action of gravity. An opening may have, for example, an annular shape (with a diameter of about 1 mm to 4 mm), a square shape (with a dimension of about 1 mm to 4 mm), an oblong shape Petition 870250090762, dated 06 / 10 / 2025, page 39 / 99 20 / 28 (with a length of about 3 to 4 mm, for example, equal to 3.50 mm, and a width of about 1 to 2 mm, for example, 1.75 mm), a rectangular shape, etc. When multiple openings are provided, each opening may have a different shape. An opening of less than 1 mm (diameter or length) may resist the free flow of liquid. An opening of more than 4 mm (diameter or length) may compromise the structural integrity of the device.

[082] In the example illustrated in figure 3B, the openings have an oblong shape, and are located at the lower end of the main section 51 in the shape of a cylinder. The length of an opening can be about 3 to 4 mm, for example, 3.50 mm, and the width can be about 1 to 2 mm, for example, 1.75 mm.

[083] According to a particular embodiment, the hollow lumen configuration 5 allows a pipette, syringe needle, or any suitable device that can be used to remove liquid from the reaction volume, or introduce liquid into the reaction volume, to be inserted into the lumen and, in particular, to extend through it into the reaction volume (e.g., parabolic end section 54).

[084] This helps introduce and remove the liquid without needing to remove the sampling device from the well or cap at any stage, thus reducing the risk of cross-contamination.

[085] Such a pipette may be, for example, a Beckman Coulter™ 230 μL Biomek™ pipette (item B85903) or an Apricot Designs™ 125 μL EZ-Load™ pipette tip, or any suitable pipette, for example, standard pipettes.

[086] This also additionally allows the use of standard liquid handling robots, such as, for example, the Beckman Coulter i7™, or any Petition 870250090762, dated 06 / 10 / 2025, p. 40 / 99 21 / 28 liquid handling machine that can handle multiple standardized plates, for example, by the Society for Biomolecular Sciences (SBS), such as Apricot Personal Pipettor™.

[087] According to one embodiment, the proximal loop portion 3 includes ribs 13. Such external ribs or fins may be provided on the proximal loop portion 3 to allow air to escape from the well, and thus facilitate the free flow of the reaction liquid through the lumen and its exit through the openings 7, while the sampling device is inserted into the well. They may also facilitate the secure and tight mounting of the device in a reaction chamber.

[088] The sampling device can be manufactured from plastic, so it can be easily manufactured by injection molding. It can be created as a single piece of plastic.

[089] For example, it can be manufactured from polypropylene or any material suitable for laboratory processing (such as liquid fluid dynamics for automatic dispensing and removal, sterilization, etc.), as well as for safe robotic operations during tissue scraping.

[090] In a particular embodiment, the sampling device is manufactured from polypropylene polymer material, such as RP373R material, i.e., polypropylene random copolymer. This material is used primarily for applications in the pharmaceutical, medical device, laboratory and diagnostic fields.

[091] In the first embodiment, the invention kit comprises the sampling device 1 described above and a well of a multi-well plate as a reaction chamber, within which the sampling device is securely mounted after a tissue sampling operation with the distal scraping part of the device nested in the base of the well. The well of the plate Petition 870250090762, dated 06 / 10 / 2025, page 41 / 99 22 / 28 multiwell plates and the scraping section of device 1 are dimensioned in order to provide a defined reaction volume between the device and the well wall for the purpose of receiving a reaction fluid. This is illustrated in greater detail in Figure 8, which shows device 1 nested in well 20 of a multiwell plate 21 and illustrates the reaction volume 16 defined by well 20 and device 1, which is supplied with reaction fluid 17 up to above the level of opening 7 to a height H.

[092] With reference to figures 4A, 4B and 5, a sampling kit, according to an alternative embodiment of the invention, is illustrated. This embodiment of the sampling device is for use with a lid as a reaction chamber and is also suitable for manual (non-automated) use. The kit comprises the sampling device 1 or an alternative sampling device 10 (which is very similar to device 1 but has a longer proximal handle part 3) and a lid 15 that functions as a reaction chamber.

[093] In a first configuration of the kit (figures 4A and 4B), the cap 15 is detachably attached to the tissue handling part 3 of the device 10 providing a handle extension for use during a tissue sampling operation. In a second configuration, used after a tissue sampling operation, the cap 15 is attached to the distal scraping part 4 of the device 10 where it acts as a reaction chamber.

[094] According to one embodiment, the sampling device 10 comprises at least one partition 41. Such partitions are located above the openings 7, for example, in an upper part of the distal scraping section. This ensures an air connection between the reaction chamber and the exterior of the sampling device when the lid is attached to the distal scraping section 4 of the device 10, and thus serves to provide ventilation of the device. Petition 870250090762, dated 06 / 10 / 2025, page 42 / 99 23 / 28 sampling.

[095] It may be noted that sampling device 1, or alternative sampling device 10, is suitable for use with an automated sampling system described below.

[096] With reference to figures 6A to 6E and 7A to 7D, the use of two embodiments of the invention kit is illustrated. In the first embodiment (figures 7A to 7D), the use of the device and kit of figures 1 to 3B is illustrated, where the reaction chamber is provided by the well of a multi-well plate. In the second embodiment (figures 6A to 6E), the use of the kit of figures 4 and 5 is illustrated, where the reaction chamber is provided by a lid 15.

[097] With reference specifically to figure 6A, the sampling device 10 and the lid 15 attached to a proximal handling end of the device are illustrated sampling tissue from a carcass surface 24. The user holds the lid 15 and scrapes the conical end and the grooves 9 along the carcass surface, or substrate, where the shallow grooves dislodge and collect some of the meat fibers without tearing or ripping pieces of the meat tissue.

[098] With reference to figure 6B, the cap 15 is then carefully detached from the proximal handling end of the device 10 and attached to the distal scraping part 4 of the device, where it serves as a reaction chamber for the cytolysis treatment of tissue fiber cells.

[099] With reference to figure 6C, the assembly is made, first with the lid, in a well 60 of a multiwell plate 21 to be stored until later analysis.

[100] With reference to figure 6D, a pipette 25 with a pipette tip 26 is used to distribute the reaction fluid to the reaction chamber in the lid 15 through the internal lumen in the device 10, without needing to detach the Petition 870250090762, dated 06 / 10 / 2025, page 43 / 99 24 / 28 lid device 15. Once the reaction fluid is dispensed, it will mix with the tissue fibers and perform cytolysis of the cells in the fiber, releasing DNA from the cells that will diffuse around the reaction fluid, including the reaction fluid in the inner lumen.

[101] With reference to figure 6E, pipette 25 with pipette tip 26 (illustrated partially by dashed lines) can then be used to remove some of the DNA-containing fluid in the inner lumen, again without needing to disassemble device 10 from cap 15.

[102] With reference specifically to figure 7A, the use of the first embodiment of the invention kit is illustrated, in which the previously identified parts are designated by the same numerical references. In this embodiment, the sampling device 1 is illustrated sampling tissue from a carcass surface 24, where the user (or a robotic arm) holds the device 1 at the proximal handling part 3 and scrapes the parabolic end with grooves 9 along the surface at an oblique angle in order to dislodge and collect the meat fibers.

[103] With reference to figure 7B, the distal scraping portion of device 1 is detachably mounted in a well 20 of a multiwell plate 21, which then serves as a reaction chamber for the cytolysis treatment of tissue fiber cells. The distal end of the device is illustrated by dashed lines to illustrate the reaction volume defined by the device and the well.

[104] With reference to figure 7C, a pipette 25 with a pipette tip 26 is used to distribute the reaction fluid to the reaction chamber in well 20 of the multiwell plate 21 through the inner lumen in the tissue sampling device 1 without needing to remove the device 1 from well 20. Once the reaction fluid is distributed, it will mix with the fibers of Petition 870250090762, dated 06 / 10 / 2025, page 44 / 99 25 / 28 tissue and will perform cytolysis of cells in the fibers, releasing DNA from the cells that will diffuse around the reaction fluid, including the reaction fluid disposed in the inner lumen. Figure 8 illustrates in greater detail device 1 in the well of the multiwell plate after the reaction fluid has been added to the well.

[105] With reference to figure 7D, pipette 25 can then be used to remove some of the DNA-containing fluid in the inner lumen, again without needing to remove the device from the well.

[106] The device and kit of the invention allow tissue to be sampled and then treated (i.e., for cell cytolysis) in a reaction chamber (i.e., a detachable lid, or a well of a multi-well plate) without needing to remove the tissue sampling device from the reaction chamber. As mentioned above, this eliminates the problem associated with prior art devices, which need to be removed from the well before the addition of the reaction liquid, causing droplets to fall onto the plate and into other wells, resulting in contamination problems. An additional advantage is the use of the device with a series of hollow circumferential grooves, which has been shown to normalize the amount of tissue removed from a carcass, regardless of whether the scraping action uses a lot or a little pressure.

[107] With reference to figure 9, an automated robotic system 30 for sampling and indexing multiple carcasses 24 with high throughput is illustrated. In this embodiment, the system comprises a multi-well plate 21 having a plurality of wells and a single barcode 40, and a plurality of tissue sampling devices 1 nested in the wells of the multi-well plate 21, each tissue sampling device having a proximal handling part 3 and a distal scraping part 4.

[108] A robotic sampling system 31 is provided and comprises a robotic arm 32 with an arm tool end 33 Petition 870250090762, dated 06 / 10 / 2025, page 45 / 99 A 26 / 28 robotic arm is configured to engage the interior of the proximal handling portion of a tissue sampling device while seated in the well of a multi-well plate. The robotic arm is configured to lift the tissue sampling device from the well, move the tissue sampling device toward a carcass and scrape the distal scraping portion against the carcass, and replace the tissue sampling device in the same well. The sampling system includes means to correlate the plate position relative to the arm, allowing the system to direct the arm to the first well in the plate, and then to the second, third, and fourth wells, and to record the well position from which the particular device was removed.

[109] The system includes a first sensor 34 which includes a camera 35 to identify, in order to detect, an electronic ID on the housing 24. The system also includes a second sensor to detect a barcode on the multiwell plate. The system also includes a processor 37 operationally coupled to the robotic system 31 and the sensors and configured to receive the detected electronic ID from the housing, the barcode from the multiwell plate, and the location of the well on the plate from which the sampling device was removed. The processor is configured to assign the electronic ID of the housing to a specific sampling device using a reference of the respective housing ID and the reference location of the well on a multiwell plate 21 and the barcode of the multiwell plate 40. All data is archived in the computer spreadsheet with a date and time stamp.

[110] The system additionally and generally includes a display system 38 for displaying the data comprising the casing ID connected to the sampling device with reference to a well location on the associated plate; and optionally, a communication system 39 for transmitting the data to a remote location via a communications system, such as Petition 870250090762, dated 06 / 10 / 2025, page 46 / 99 27 / 28 the Internet.

[111] The system can be used to collect tissue samples from multiple carcasses in a slaughterhouse, for example, up to 10,000 per day, which are stored in multi-plate wells of 96 wells. The system provides data comprising the IDs for the carcass samples and for each carcass data relating to the specific well, and to the plate on which the tissue sample from that carcass was stored. The data can be displayed on paper using a printer or recorded on another medium, such as a disk. The data can also be electronically transferred via a communication system to the laboratory where the samples are to be analyzed.

[112] Although not illustrated, the system may include a liquid handling robot configured to simultaneously dispense the reaction fluid into a plurality of wells of a multi-well plate, and simultaneously remove a quantity of a reaction mixture, or of a product produced by the reaction mixture, from the plurality of wells of the multi-well plate. The robotic liquid handling part of the system is generally located in a different location from the robotic sampling system, for example, in a laboratory. In one embodiment, the outer base of each of the wells of the multi-well plate includes an electronic identification indicator (e.g., a barcode) readable by a flatbed scanner.

[113] The liquid removed from the reaction chamber is usually treated to analyze the DNA profile or genotype or to identify single nucleotide polymorphisms or other polymorphisms or haplotypes in the DNA. This may involve DNA multiplexing amplification and subsequent next-generation sequencing of all or part of the DNA. High-throughput or next-generation sequencing methods and kits are available, Petition 870250090762, dated 06 / 10 / 2025, page 47 / 99 28 / 28 including Synthesis Sequencing (Illumina), Pyrosequencing (454), Ion Semiconductor Sequencing (Ion Torrent Sequencing) and Combinational Probe Anchor Synthesis (cPAS-BGI-MGI). Targeted NGS is a next-generation sequencing technique that focuses on specific amplicons and genes and employs amplification of the required gene or amplicon by enzymatic amplification, which is then sequenced on an NGS platform. It is described in Bybee et al (Targeted Amplicon Sequencing (TAS): A Scalable Next-Gen Approach to Multilocus, Multitaxa Phylogenetics. Genome Biology and Evolution. 3: 13121323. doi: 10.1093 / gbe / evr106. PMC 3236605) and Masser et al (Targeted DNA Methylation Analysis by Next-Generation Sequencing). The fluid removed from the chamber can be analyzed to detect quantitatively or qualitatively other tissue components, for example, other nucleic acids or proteins. Equivalencies

[114] The above description details the currently preferred embodiments of the present invention. Numerous modifications and variations in practice, therefore, must occur to those skilled in the art upon consideration of these descriptions. These modifications and variations must be encompassed by the appended claims. Petition 870250090762, dated 06 / 10 / 2025, page 48 / 99

Claims

1 / 3 CLAIMS 1. Sampling kit comprising a reaction chamber (15, 20) and a sampling device (1, 10) configured to be mounted in the reaction chamber and to provide a defined reaction volume (16) between the base of the reaction chamber and the sampling device, the sampling device having a proximal handling part (3) and a distal scraping part (4) comprising a scraping formation (9) configured to collect a tissue sample when the distal scraping part is rubbed against a tissue surface, characterized in that the sampling device comprises an inner lumen (5), extending from an open top to at least one opening in a side wall of the distal scraping part,the said lumen being configured to supply the reaction fluid from a distal end of the tissue sampling device to the reaction volume through a distal opening (7) disposed in the distal scraping part, when the tissue sampling device is mounted in the reaction chamber.

2. Sampling kit according to claim 1, characterized in that the reaction chamber comprises a lid (15) for the sampling device (10).

3. Sampling kit, according to claim 1, characterized in that the reaction chamber comprises a well (20) of a multi-well plate (21).

4. Sampling kit, according to any one of claims 1 to 3, characterized in that the sampling device (1, 10) is hollow.

5. Sampling kit, according to any one of claims 1 to 4, characterized in that it includes at least two distal openings (7) in the distal sampling part. Petition 870260070160, dated 15 / 07 / 2026, page 12 / 19 2 / 3 6. Sampling kit, according to any one of claims 1 to 5, characterized in that the distal opening (7) in the distal sampling part (4) is arranged close to the scraping formation (9).

7. Sampling kit, according to any one of claims 1 to 6, characterized in that the inner lumen is configured to receive a pipette tip or a syringe needle.

8. Sampling kit, according to any one of claims 1 to 7, characterized in that the distal scraping part comprises a parabolic end (8).

9. Sampling kit, according to any one of claims 1 to 8, characterized in that the scraping formations (9) comprise one or more circumferential grooves.

10. Sampling kit, according to claim 9, characterized in that one or more circumferential grooves have a depth of not more than 0.5 mm.

11. Sampling kit, according to any one of claims 1 to 10, characterized in that the scraping formation comprises scraping edges configured to collect the fibers of the meat tissue.

12. Sampling kit, according to any one of claims 1 to 11, characterized in that the sampling device is manufactured as a single piece of plastic.

13. Sampling kit, according to any one of claims 1 to 12, characterized in that the sampling device (1, 10) and the reaction chamber (15, 20) are configured to be assembled together by press fitting.

14. Sampling kit, according to any one of claims 1 to 13, characterized in that it includes a plurality of sampling devices (1), each being detachably mounted in a well (20) of a multi-well plate (21).

15. Sampling kit, according to any one of claims 1 to 14, characterized in that the sampling device comprises an open proximal end (6) and in which the internal lumen (5) is configured to supply the reaction liquid from the open proximal end (6) to the reaction volume. Petition 870260070160, dated 15 / 07 / 2026, p. 14 / 19

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