Apparatus suitable for preparing a sample for analysis

Abstract

An apparatus 100 suitable for preparing a sample for analysis comprises a body 101 defining a cavity 102 with an entrance aperture 103, a seal 104 arranged on the body to, in use, engage with a container 108 and form a liquid tight seal between the container and the body, a valve 105 arranged in fluid communication with a filter 106 and configured to prevent fluid contained within the body from passing through the entrance aperture 103; the filter is arranged in fluid communication with the entrance aperture. The body has an egress aperture 109, the body may comprise a movable body and have an absorber in the body. An apparatus may comprise a body with first and second arms with fluid collector and an absorber. An apparatus may comprise a body defining a cavity with entrance aperture, a rupturable seal occluding the aperture, and a captured filter arranged to rupture the seal.

Description

Field The present invention generally relates to an apparatus, kit, system, and method of use of apparatus, suitable for preparing a sample for analysis. Introduction Sample preparation and analysis traditionally takes place in the controlled environment afforded by a laboratory by a dedicated technician typically having numerous years of experience in such matters. This necessarily involves a high cost and time delay in the provision of such facilities, which are normally located remote from where a sample was taken or a customer who required such a sample to be analyzed. The safe collection of fluid samples is problematic, as such samples may require the sample is kept at a controlled temperature to ensure a valid analysis (e.g. if containing a live culture). Collection of such samples may be hazardous in itself, where such samples may be a hazardous substance or simply difficult to obtain directly (e.g. from livestock). The sample, once collected, must then be transported, at cost and in the required manner (e.g. at a controlled temperature) to the laboratory for analysis. One or more embodiments of the present invention aim to solve one or more of the above mentioned problems. In particular, removing the need for a dedicated laboratory and technician, or the controlled transport of a sample to a remote facility. Statements of Invention The inventors have developed a novel apparatus, kit, system, and method of use of apparatus, suitable for preparing a sample for analysis. The novel apparatus, kit, system, and method facilitate sample collection an analysis in the field by non-expert users or lay people. Such samples which may be collected include, but are not limited to bodily secretions, milk, blood, urine, saliva, and the like. Advantageously, this reduces expense and time of performing, and obtaining results from, tests in the field. Apparatus in fluid communication with a filter In one aspect of the present disclosure, the invention provides an apparatus, suitable for preparing a sample for analysis, the apparatus comprising: a body defining a cavity with an entrance aperture; a seal arranged on the body to, in use, engage with a container and form a liquid tight seal between the container and the body; a valve arranged in fluid communication with a filter and configured to prevent fluid contained within the body from passing through the entrance aperture; and wherein the filter is arranged in fluid communication with the entrance aperture. Suitably, the apparatus may include, or forms part of, a filter unit, thereby allowing a fluid to be filtered during collection and ensuring impurities are removed from the fluid prior to analysis. Suitably, the seal may be an O-ring, such as a rubber or silicone O-ring or the like, such that a fluid-tight seal may be achieved. The valve may be an umbrella valve, such that fluid is permitted to flow in a single direction. Suitably, the valve may be arranged adjacent the filter, such that filtered fluid may not pass back through the filter and meet the unfiltered fluid. Suitably, the fluid may be a sample. The sample being collected from a target that a user wishes to obtain some information about. Samples which may be collected include, but are not limited to bodily secretions, milk, blood, urine, saliva, and the like from any living creature including humans, animals, including livestock. Suitably, the sample may be milk. Suitably, the filter may be arranged adjacent the entrance aperture, such that the fluid must first pass the filter, ensuring the apparatus is not contaminated by impurities which may be caught by the filter. Suitably, the body comprises an egress aperture arranged in in fluid communication to the entrance aperture, the egress aperture providing an exit for a prepared fluid to exit the apparatus for further preparation or analysis of the fluid. Suitably, the egress aperture may be arranged in opposition to the entrance aperture. Suitably, the egress aperture may be provided with a temporary closure such that the fluid may not pass through the egress aperture unless and until the temporary closure is removed or ruptured. Suitably, the temporary closure may be a cap (such as a push or screw cap), film or foil, and further optionally or alternatively wherein the temporary closure is peelable from the apparatus, such as by hand or absent a tool. Suitably, the cavity comprises a rehydration buffer solution. In this way the apparatus may be pre-primed to conduct partial or full preparation of a fluid introduced to the apparatus. Suitably, at least a portion of the body may be dimensioned complementary to an internal dimension of a container, such that insertion of the body into the container forces a fluid contained within the container through the filter, valve, and into the cavity, optionally wherein the container is a sample processing container. In this way, the portion of the body dimensioned complementary to an internal dimension of a container may act as a piston within the container and form a fluid tight seal with the container. Suitably, the body may be urged towards the container, and the fluid forced through the filter, by hand, without additional tooling. Suitably, the container may comprise a fluid absorber, the fluid absorber arranged to absorb fluid and to then release the fluid upon insertion of the body into the container. Such a fluid absorber can make samples easier to handle by retaining the fluid within the container, even if the container is, for example, inverted. The absorber prevents the fluid leaving the container until a user intentionally withdraws the fluid from the absorber, by, for example, compressing the absorber and thereby displacing the fluid. Suitably, the absorber, in use, may provide a filter and retains impurities such as solid particulate matter held within the sample. The absorber may provide a riddle or maze by which impurities are trapped, or such impurities may preferentially adhere to the absorber rather than remain in the fluid. Suitably, the absorber may be pre-treated with a reagent. In this way the apparatus may be pre-primed to conduct partial or full preparation of a fluid introduced to the apparatus. Suitably, a kit of parts comprising the apparatus described above may be provided, further including at least one container, such as a sample processing container, and the kit further optionally comprising one or more from the list: a fluid, a liquid, a rehydration buffer, and / or a cap. The kit may suitably comprise a cap, wherein the cap is configured to occlude the entrance aperture in order to prevent ingress of contaminants or to retain the fluid within the apparatus. Suitably, a system comprising an apparatus as described above may be provided, and further including a container, such as a sample processing container, and the system further optionally comprising one or more from the list: a fluid, a liquid, a rehydration buffer, and / or a cap. The system may further comprise a cap, wherein the cap is configured to occlude the entrance aperture in order to prevent ingress of contaminants or to retain the fluid within the apparatus Apparatus comprising an absorber In one aspect of the present disclosure, the invention provides an apparatus, suitable for preparing a sample for analysis, the apparatus comprising a body defining a cavity with an entrance aperture and an egress aperture arranged in fluid communication with the entrance aperture; wherein the body comprises a movable member, and an absorber associated with the body. Such a fluid absorber can make samples easier to handle by retaining the fluid within the container, even if the apparatus is, for example, inverted. The absorber prevents the fluid leaving the container until a user intentionally withdraws the fluid from the absorber, by, for example, compressing the absorber and thereby displacing the fluid. An absorber can be sized to absorb a predetermined volume of fluid, thereby simplifying collection of the correct volume of a fluid. Suitably, the apparatus may include, or form part of, a filter unit, thereby allowing a fluid to be filtered during collection and ensuring impurities are removed from the fluid prior to analysis. Suitably, the egress aperture may be arranged opposite the entrance aperture. Suitably, the absorber, in use, provides one or more of a wick, a filter, and / or is sized to absorb a predetermined volume of a fluid. The absorber may therefore remove impurities and also simplify sample collection. The absorber may perform the aforesaid functions singularly or together. Suitably, the egress aperture is adapted to interface with a fluid extractor, such that the fluid, once absorbed by the absorber, may be extracted for further processing or analysis directly from the apparatus. Suitably, the movable member may be displaceable within the cavity between an entrapping position and a releasing position. Suitably, the movable member may be placed in either the entrapping position or the releasing position by depressing the egress aperture. Depressing the egress aperture should be achievable by hand and without the need of extra tooling. Suitably, the movable member may be arranged to entrap a reagent when in the entrapping position, and to release the reagent when in the releasing position. In this way the movable member may be used to retain an element that can be released at will by a user of the apparatus. Suitably, the reagent may comprise a lysing agent, such as a lyophilized agent, optionally a bacteriocin, further optionally wherein the bacteriocin is lysostaphin. Suitably, the reagent may be contained on a bead. The bead may be a lyophilised bead. Suitably, the body may be arranged to engage, and fluidly seal with a container, with the wick arranged within the container, this allows for fluid communication between a container and the wick without ingress or egress of fluid about the body. Suitably, a kit of parts comprising the apparatus described above may be provided, and further comprising a container, such as a sample processing container, and the kit further optionally comprising one or more from the list: a fluid, a liquid, a rehydration buffer, a pouch, a flexible container, a syringe, and / or a cap. The kit may further optionally comprise a fluid. The kit may further optionally comprise a liquid. The kit may further optionally comprise a rehydration buffer. The kit may further optionally comprise a pouch. The kit may further optionally comprise a flexible container. The kit may further optionally comprise a syringe. The kit may further optionally comprise a cap. When comprising a cap, suitably the kit, is configured to occlude the egress aperture, via the cap. Suitably, a system comprising an apparatus as described above may be provided, and further including a container, such as a sample processing container, and the kit further optionally comprising one or more from the list: a fluid, a liquid, a rehydration buffer, a pouch, a flexible container, a syringe, and / or a cap. The system may further comprise a cap, wherein the cap is configured to occlude the egress aperture. Apparatus comprising arms In one aspect of the present disclosure, the invention provides an apparatus, suitable for preparing a sample for analysis, the apparatus comprising: a body having first and second arms, the first arm bearing a fluid collector the second arm bearing an absorber. Such a fluid collector can make collecting samples easier, providing a mechanical filter where particulate matter in the fluid is unable to enter the collector, e.g. via a weir or suitably dimensioned aperture. An absorber can be sized to absorb a predetermined volume of fluid, thereby simplifying collection of the correct volume of a fluid. The apparatus may also be provided as the body having only the first arm bearing the fluid collector and absent the second arm. Suitably, the fluid collector may comprise a reagent, wherein the reagent optionally comprises a lysing agent, optionally a bacteriocin, further optionally wherein the bacteriocin is lysostaphin. Suitably, the reagent may be contained on a bead. Suitably, the reagent may be contained within a rupturable chamber, such that the reagent may only be released once a user ruptures the rupturable chamber at the desired time. Suitably, the absorber may be pretreated with a reagent, wherein the reagent optionally is a buffer. Suitably, the absorber may be sized to absorb a predetermined volume of fluid, and optionally or alternatively, the absorber is arranged to release an absorbed fluid. An absorber can be sized to absorb a predetermined volume of fluid, thereby simplifying collection of the correct volume of a fluid without additional fluid measuring devices. Suitably, a kit of parts comprising the apparatus described above may be provided, and further comprising one or more of a container; a rupturer; a cap. The kit may suitably comprise a rupturer, wherein the rupturer is sized to accept the fluid collector, and the rupturer arranged to rupture the rupturable container. Suitably, the rupturer may comprise a piercing element in order to rupture the rupterable container. Suitably, the rupturer may comprise a fluid, and optionally wherein the volume of fluid is defined relative to a fluid volume capacity of the fluid collector. Suitably, the cap may be sized to engage with the body and shield the absorber in order to protect the absorber from contamination or from prematurely releasing the fluid. Suitably, a system comprising the apparatus described above may be provided, and further comprising one or more of a container; a rupturer; a cap. Suitably, the rupturer of the system may be sized to accept the fluid collector, and the rupturer arranged to rupture the rupturable container. Suitably, the rupturer may comprise a piercing element. Suitably, the rupturer may comprise a fluid, and optionally wherein the volume of fluid is defined relative to a fluid volume capacity of the fluid collector. Suitably, the cap is sized to engage with the body and shield the absorber. Apparatus comprising a captured filter In one aspect of the present disclosure, the invention provides an apparatus, suitable for preparing a sample for analysis, the apparatus comprising: a body defining a cavity with an entrance aperture; a rupturable seal occluding the entrance aperture; a captured filter arranged adjacent the rupturable seal; wherein the captured filter is arranged to rupture the rupturable seal and thereby establish fluid communication between the captured filter and the cavity. Suitably, the captured filter is associated with the body. The captured filter need not be connected, directly, or indirectly, to the body. Suitably, one or more seals are arranged on the body and / or the captured filter to, in use, engage with a container and form a liquid tight seal between the container and the body and / or filter. Suitably, the captured filter comprises a first filtration stage and a second filtration stage, thereby providing complete filtration without additional filters. The fist and second filtration stages may be achieved by layers of discrete filters stacked one on another. Suitably, a secondary filter is provided. The secondary filter may be provided, as an alternative to the captured filter comprising first and second filtration stages. The secondary filter may be provided at any suitable location within the apparatus, provided it may fulfil its function of providing a further filtration of a fluid within the apparatus. Suitably, the captured filter is slidably engaged with the body, and optionally wherein sliding the captured filter from a first position to a second position causes the rupturable seal to rupture. The captured filter maybe provided with a rupture to facilitate breaking the seal. Suitably, the captured filter is urge-able towards the body, and optionally wherein urging the captured filter towards the body causes the rupturable seal to rupture. Such urging may cause rupture of the seal by changes in fluid pressure to in excess of a limit tolerable by the seal or of an adhesive connecting the seal to the body. Suitably, the cavity comprises a variable volume, optionally wherein the cavity may transition from a first volume to a second volume. Suitably, a fluid located within the cavity is caused to flow by variation of the cavity volume. Suitably, the volume of the cavity is varied by a piston. The piston may be provided in the form of a syringe, built integrally into the body and in fluid communication with the cavity. Suitably, the cavity comprises a rehydration buffer solution. Suitably, at least a portion of the body, and / or captured filter, is dimensioned complementary to an internal dimension of a container, such that insertion of the body, and / or captured filter, into the container causes rupture of the rupturable seal. Suitably, a kit of parts comprising the apparatus described above may be provided. The kit of parts comprising an apparatus as above, and at least one container, such as a sample processing container, and the kit further optionally comprising one or more from the list: a fluid, a liquid, a rehydration buffer, and / or a cap. Suitably, a system comprising the apparatus described above may be provided. The system comprising an apparatus as described above, and a container, such as a sample processing container, and the system further optionally comprising one or more from the list: a fluid, a liquid, a rehydration buffer, and / or a cap. Kit In one aspect of the present disclosure, the invention provides a kit of parts for collecting, filtering, and preparing a sample for analysis, the kit comprising: a container; an apparatus as discussed in any of the aspects disclosed above; and a processing container. Suitably, the kit optionally further includes a heater and / or an analyser, optionally wherein the analyser comprises a further filter. Suitably, the container may comprise an aperture sized to allow collection of 20-100 ml of a fluid, optionally collection directly from a living creature or processing line, and further optionally wherein the container comprises an indicator marker. Methods of using the Apparatus In one aspect of the present disclosure, the invention provides a method of preparing a sample for analysis, using a kit as discussed above, the method comprising: collecting a sample in the container; treating the sample with a reagent; adding a buffer solution to the sample; heating the sample to a first temperature for a first period of time, heating the sample to second temperature for a second period of time, and cooling the sample to ambient temperature; and handling the sample with the apparatus. Suitably, the below steps may be performed in the following order: (a) collecting a sample in the sample container; (b) treating the sample with a reagent; (c) adding a buffer solution to the sample; (d) heating the sample for a first period of time, heating the sample for a second period of time, and cooling the sample to ambient temperature. Suitably, the method may further comprise the step of analyzing the filtered sample. Suitably, the reagent may comprise a lysing agent, optionally a bacteriocin, further optionally wherein the bacteriocin is lysostaphin. Suitably, the first temperature is 37°C and the second temperature is 100°C. Figures Figure 1 shows a cross section of an apparatus; Figure 2 shows a cross section of an apparatus; Figure 3 shows a cross section of an apparatus; and Figure 4 shows a cross section of an apparatus. The invention will now be further described with reference to the following non-limiting examples, and the figures described above. Examples Apparatus in fluid communication with a filter Figure 1 shows a cross section of an apparatus (100) suitable for preparing a sample for analysis. The apparatus has a body (101) defining a cavity (102) with an entrance aperture (103); a seal (104) arranged on the body (101) to, in use, engage with a container and form a liquid tight seal between the container and the body (101); a valve (105) arranged in fluid communication with a filter (106) and configured to prevent fluid contained within the body from passing through the entrance aperture (103); and wherein the filter (106) is arranged in fluid communication with the entrance aperture (103). The container can be a tube (108) comprising a sample to be analyzed, typically a fluid. Suitable tubes (108) include, but are not limited to, a polypropylene tube (108). The tube (108) may also take the form of a cap to provide a mechanical cover for the entrance aperture (103), protecting the apparatus (100) from ingress of contaminants or other materials. In use, the apparatus (100) is inserted into the tube (108) and the user applies a pressure, urging the apparatus into the tube (108) and thereby forcing the fluid contained within the tube (108) through the entrance aperture (103), through filter (106) and the valve (105) and into the cavity (102). The valve (105), acts as a non-return valve preventing the fluid from returning through the filter (106) and the entrance aperture (103), and thereby providing a filtered fluid, ready for further preparation or analysis in the cavity (102), the apparatus thereby forms, or includes a filter unit. A suitable example filter has a density of 0.07 g / cc. Other densities may be suitable such as up to 0.1 g / cc or 0.15 g / cc. The valve also prevents any pre-loaded reagent (107), where present, from leaking out of the apparatus (100) during storage and handling. In some cases, a secondary filtration step may also be required to further purify the fluid and / or to prevent blockage of analytical instruments. A secondary filter (not shown) may be placed elsewhere within the apparatus between the filter (106) and the egress aperture. The secondary filter may be placed inside the apparatus, such as within the cavity (102) towards an egress aperture (109) or alternatively, on top, or adjacent, the filter (106), prior to the valve (105). The egress aperture (109) provides an exit for a prepared fluid to exit the apparatus for further preparation or analysis of the fluid. The cavity may be supplied with a reagent (107) to which the fluid is introduced when the apparatus (100) is urged into the tube (108). Including a reagent (107) means this arrangement simultaneously filters the fluid and mixes the fluid with the reagent (such as, but not limited to a rehydration buffer). The apparatus (100) is sized complimentary to the internal diameter of the tube (108) in order to provide a tight seal between the inside of the tube (108) and the outside of the filter unit. A seal (104), such as, but not limited to, an O-ring may be placed adjacent the entrance aperture (103). Any appropriate seal may be used provided the seal serves the purpose of preventing the fluid in the tube (108) flowing between the outside diameter of the apparatus (100) and the internal diameter of the tube (108), thereby ensuring the fluid passes through the entrance aperture (103). The reagent (107) may be provided in a predetermined volume, sized to be sufficient to interact with a fluid entering the cavity (102) to achieve a desired outcome. An example volume of reagent may be in the range 100 pL to 500 pL, in particular. The volume of reagent (107) may, for example, be sized to achieve a ratio of 2 part fluid to 3 parts reagent by volume, following the passage of the fluid through the filter (106). The apparatus may be sized to yield is approximately 150 uL of filtered fluid from an initial fluid volume of 600 pL. When the 150 uL of filtered fluid mixes with the reagent (107) the final fluid volume is under 400 pL. The egress aperture (109) is provided with a temporary closure (110) such that the fluid may not pass through the egress aperture (109) unless and until the temporary closure (110) is removed or ruptured. The temporary closure (110) may be a film or foil, and further optionally or alternatively wherein the temporary closure (110) is peelable from the apparatus, such as by hand or absent a tool. The user removes the temporary closure (110) which covers the egress aperture. This action may also releases an elevated air pressure within the cavity (102) which may have formed during urging the apparatus (100) into the tube (108). The filtered fluid, mixed with the reagent (107, where present), may then be removed from the apparatus (100) and then further user, analysed, and / or treated, such as, by way of non-limitative example, loaded onto an analysis cartridge by, such as, either pouring, or by using a pipette or syringe. The above apparatus is applicable to the preparation of clinical mastitis milk samples in the field. Such samples can be very variable in consistency depending on the host immune response and the causative pathogen. As a result, the physical characteristics of each sample and the consistency of the sample (including after initial preparation, including heating) vary greatly. Filtration to remove lumps and debris from the milk samples after heating allows the sample fluid to then be analysed. Some fluids can solidify on initial processing (e.g. milk samples can solidify following a high heat incubation treatment). Subsequent handling of such samples is difficult and often results in protocol failure. A solution to this issue is to provide a fluid absorber (not shown) in the tube (108). The fluid absorber is arranged to absorb fluid and to then release the fluid upon insertion of the body (101) into the container (tube 108) through compression of the absorber by the body (101). The absorber can prevent the solidification of some samples upon treatment (milk samples following a high heat incubation treatment, for example). Where the initial preparation required an initial reagent to be mixed with the fluid, the absorber can hinder the mixing process. Therefore, the fluid may need to be pre-mixed with the initial reagent prior to introduction of the fluid to the absorber. Where this is the case, the container may be pre-supplied with a given volume of the initial reagent (such, as a buffer like IDTE) and then user introduces the fluid. The container may be provided with a fill marker to ensure a known quantity of fluid is added to the container. The fill marker may be arranged to ensure a particular a ratio of fluid to initial reagent, such as, two parts fluid and one part initial reagent, by way of non-limitative example. The absorber can be introduced to the same container, or a volume of fluid transferred to another container pre-supplied with the absorber. The size of the absorber may be selected to ensure a majority of the fluid is absorbed. The size of the absorber may be selected such that a pre-determined volume of fluid is absorbed. The absorber may provide a filter function and retain impurities such as solid particulate matter held within the fluid. A non-limited example use of the above apparatus may be as follows: A milk sample is collected in a generic collection vessel with ideal volume between 20 and 100 mL, to allow for easier handling. The diameter of the vessel may be relatively large (such as approximately 5-10 cm), to allow for easier sample collection directly from a lactating animal or parlor line. The container may be provided with a marker, such as a ‘fill-to-line’, to ensure users do not overfill this container. The volume collected at this stage is not essential, as long as it is sufficient to allow for a sub-sample to be taken. A small sub-sample is aspirated out of the generic collection vessel, such as by using a 1 mL syringe. A larger capacity syringe could be used, however this will reduce accuracy. Once the milk sample is aspirated into the syringe the sample is mixed with a lysostaphin bead. The lysostaphin bead will be rehydrated and the enzyme will dissolve into the milk sample. The volume of the sample collected here is 400 pL. However this volume can be scaled in line with the volumes of other reagents, to better match the user case and / or cost requirements of the product. The user then dispenses the whole volume (400 uL) into a container (108) (a suitable tube such as a 2 mL tube) which is pre-filled with 200 pL of IDTE (1XTE buffer, pH 8.0). The action of dispensing the milk sample into the IDTE allows for some mixing of sample, IDTE and lysostaphin to occur, however it is advisable for the user to invert the container (108) a few times to ensure reagents are mixed adequately. The user places the container (108) into a sample handling unit (SHU); this is a heating and cooling dry bath unit which is pre-programmed to incubate the sample at 37°C for 10 minutes, followed by heating at 100°C for 15 minutes. The SHU will then cool down the sample to an acceptable temperature close to room temperature (20°C). The SHU takes some time to ramp up and down during incubation, and actual incubation times at the given temperatures can be further adjusted (primarily reduced) to account for the time spent ramping, in order to reduce total processing time. At this stage, the user will fit the body (101) into the container (108) and apply pressure in order to simultaneously filter the incubated sample and mix the sample with rehydration buffer (107, RB), which is already pre-loaded into the body (101). The body (101) is of exact size which allows a tight seal between the inside of the container (108) and the outside of the body (101); this is further assisted by an O-ring which is placed on the body (101). The body (101) also includes a filter (106), which removes lumps and debris from the filtrate, and a valve which prevents the pre-loaded RB from leaking out of the body (101) during storage and handling. The volume of RB is measured to be 225 pL, in order to achieve a ratio of 2 part sample to 3 parts RB following filtration. The approximate recoverable volume from filtering a 600 pL diluted and heated sample using the body (101) is approximately 150 uL, and the final lysate is under 400 pL in volume. The user then removes the temporary closure (110, foil) which covers the top of the body (101), which may release some air pressure. The lysate is then loaded onto an analysis cartridge by either pouring, or by using a pipette or syringe. Where an absorber is present in the container (108), the above method may be varied in that the syringe is used to take up a volume of 600 pL of diluted milk sample which is dispensed into the container (108) comprising the absorber (such as a wick e.g. 8.5mm in diameter, 0.1g / cc in density, cut to approximately 1.5cm in length, optionally having a hollow core). The diluted sample is absorbed into the absorber. The body (101) is then pressed into the container (108), until a shoulder of the body (101)engages with the entrance aperture of the container (108) preventing further insertion of the body (101) into the container (108), or until the absorber may not be further compressed by urging the body (108) into the container (108). This action completed the filtration of the sample and mixed the rehydration buffer (RB) with the filtered sample, resulting in a total lysate volume of approximately 500 pL. This varies slightly from one sample to the next, with more challenging samples producing less total lysate volume. The container (108) with the sample are then incubated in the SHU as above. Following incubation, the body (108) is used to filter the sample and mix the lysate with RB. The volume of sample and RB can be adjusted as needed, as long as their relative ratio is maintained. The size of the absorber also needs to be adjusted to ensure the majority of the sample is absorbed by the absorber. Apparatus comprising an absorber Figure 2 shows a cross section of an apparatus (200), suitable for preparing a sample for analysis, the apparatus (200) comprising a body (201) defining a cavity (202) with an entrance aperture and an egress aperture arranged in fluid communication with the entrance aperture; wherein the body (201) comprises a movable member (203), and an absorber (204) associated with the body (201). The absorber (204) is compressible and attached to the body (201) via a moveable member (203). The purpose of the absorber is to absorb a fluid, thereby collect a sample for processing. The moveable member (203) displaceable within the cavity (202) between an entrapping position and a releasing position. The movable member (203) may be placed in either the entrapping position or the releasing position by depressing the egress aperture. The movable member (203) is arranged to entrap a reagent when in the entrapping position, and to release the reagent (206) when in the releasing position. In this way the movable member may be used to retain an reagent that can be released at will be a user of the apparatus. The reagent (206) is anything that a user wishes to introduce to a fluid once collected by the absorber, and the fluid released from the absorber. The reagent (206), may, for example, be immobilized on a bead. The reagent (206) may be, for example, lysing agent, optionally a bacteriocin, further optionally the bacteriocin is lysostaphin, or provided as a lysostaphin bead. The reagent may also be a fluid or other solid preparation. The egress aperture is provided with a tip (205) to allow the apparatus (200) to interface with further devices. The tip (205), may be, for example, a Luer™ compatible tip, or any other similar tip (205) which facilitates aspiration of the sample from the absorber onto a syringe. A seal such as, but not limited to, an O-ring may be placed between the movable member (203) and the body (201). Any appropriate seal may be used provided the seal serves the purpose of preventing the fluid flowing between the outside diameter of the movable member (203) and the internal diameter of the body (201). The seal may also serve to provide an improved interference fit between the moveable member (203) and the body (201), such that the movable member (203), and the reagent (206) entrapped within, may only be released via a determined, deliberate movement of the movable member (203) by a user, thereby preventing accidental, or premature, release of the reagent (206). The apparatus is used by introducing the absorber (204) by the user into a fluid to be sampled. A measured volume of fluid is absorbed. The absorber may be dimensioned to take up a predetermined volume of fluid. The collection volume can be controlled by altering the diameter and length of the absorber. The user then places the absorber inside a soft pouch which may also contains a measured volume of an initial reagent, (such as, but not limited to a buffer, such as I DTE). The pouch may be sealed by mechanically attaching the body (201) onto the pouch opening via complimentary fixings, such as a screw and thread, bayonet fixing, or the like. Any suitable fixing may be used provided a fluid tight seal between the body (201) and the pouch may be achieved. The user presses down on the egress aperture, placing the movable member (203) into the release position, which releases the reagent (206) into the pouch. The pouch is then compressed multiple times to trigger a mixing action between the reagent (206) and the fluid. This action can be carried out by either the user (manually), or mechanically via a sample handling unit. The apparatus is suitable for receiving applied heat, for example, via incubation and formed of suitable materials (such as polypropylene). For example, the fluid in the pouch with the attached apparatus (200) may be incubated at a variety of temperatures without affecting the function of the apparatus (200) (e.g. 37°C for 10 min, 100°C for 15 min and then cooled down to 20°C). The tip (205) may be provided with a protective cap (208) to allow such manipulation of the apparatus (200) when attached to a pouch or otherwise. The cap (208) prevents ingress of contaminants and egress of the fluid within the apparatus (200) until the user desires. A non-limited example use of the above apparatus may be as follows: A milk sample is collected in a generic collection vessel with ideal volume between 20 and 100 mL, to allow for easier handling. The diameter of the vessel may be relatively large (such as approximately 5-10 cm), to allow for easier sample collection directly from a lactating animal or parlor line. The container may be provided with a marker, such as a ‘fill-to-line’, to ensure users do not overfill this container. The volume collected at this stage is not essential, as long as it is sufficient to allow for a sub-sample to be taken. The absorber (204) of the apparatus (200) is then is dipped by the user into the milk collection container, and a measured volume of milk is absorbed into the absorber. The collection volume can be controlled by altering the diameter and length of the absorber. The user places the absorber inside a soft pouch which contains a measured volume of I DTE buffer as per the example method above, and seals it by connecting (e.g. via complementary threads) the body (201) onto the pouch opening. The user presses down on the movable member (203), which releases the lysostaphin bead (206) into the pouch. The pouch is then compressed multiple times to trigger a mixing action between the RB and the milk sample. This action can be carried out by either the user (manually), or the SHU (sample handling unit). The sample is then incubated at the required temperatures (37°C for 10 min, 100°C for 15 min) and cooled down to 20°C. Following incubation, a syringe prefilled with RB is attached to the tip (205) of the body (201) and the lysate is aspirated out of the absorber, while being mixed with the measured volume of RB. The lysate is now loaded onto the microfluidics cartridge using the syringe. As secondary filter may be provided within the flow path of the lysate, such as within or adjacent the syringe. The secondary filter may be provided as an external accessory or fitted (optionally integrally) or within the syringe. The volume of IDTE, RB and the dimensions of the wick can easily be adapted to ensure the optimal ratios of sample to IDTE, sample to RB are maintained, and the protocol is scalable. Apparatus comprising arms Figure 3 shows a cross section of an apparatus (300), suitable for preparing a sample for analysis, the apparatus (300) comprising: a body (301) having a first arm (302) and a second arm (307), the first arm (302) bearing a fluid collector (303) the second arm bearing an absorber (304). A cap (305) may be provided to protect the absorber from contamination. The absorber (304) may be pre-treated with an initial reagent, such as those previously discussed. The absorber (304) is made of a material capable of releasable holding to fluid, such as a wicking material. This applies to the other absorbers mentioned in this description too. The fluid collector may comprise a reagent (306), similar to the reagents discussed previously, that is, anything that a user wishes to introduce to a fluid once collected by the absorber, and the fluid released from the absorber. The reagent (306), may, for example, be immobilized on a bead. The reagent (306) may be, for example, lysing agent, optionally a bacteriocin, further optionally the bacteriocin is lysostaphin, or provided as a lysostaphin bead. The reagent may also be a fluid or other solid preparation. The reagent may be entrapped within a rupturable chamber within the fluid collector (303). The rupturable chamber may comprise a temporary closure (308) such that the reagent (306) may not be released, unless and until the temporary closure (308) is ruptured. The temporary closure may be a film or foil. The user collects a fluid (309) (such as a milk sample) in a container. The fluid collector (303) is introduced into the fluid, and thereby collects part of the fluid. Any solid particulates or lumps remain in the container as these tend to be denser than the fluid and sink to the bottom of the container. That said, in some cases inclusion of some particulate matter or lumps into the fluid collector (303) may be desirable, as such matter may comprise a higher concentration of an analyte the fluid alone. The solids / lumps may be dissolved or suspended in solution during processing of the fluid sample. A seal (308) (such as those previously discussed, e.g. an O-ring) may be included to prevent the fluid from contacting the reagent (306) or the reagent may be maintained separately within an isolated rupturable chamber. Once the fluid is collected, the body (301) is urged into a rupturer (310) which carries a predetermined volume of an initial reagent (312), which is as previously discussed. The rupture is a vessel that comprises rupturing means (311) to pierce the temporary closure (310) and thereby release the reagent (306). Where the fluid (309) is separated from the reagent in the fluid collector, the process of urging the body (301) into a rupture (310) may also result in the release of the fluid (310) into the rupturer (310). The rupturing means may take the form of a piecing element arranged towards a base of the rupture (310). The volume of the initial reagent (312) may be sized to be in ratio with the carrying capacity of the fluid collector (303), such as, but not limited to, to ensure a 2 part fluid to 1 part initial reagent ratio. The user can move the body (301) in a circular motion or ‘pump’ the body (301) up and down within the rupturer (310) to mix the initial reagent (312), fluid (309) and reagent (306). The rupture (310) can be separated from the body (301) and be used to further process the fluid, such as via incubation. When ready for further processing, a user may utilize the absorber (304), which may be protected by a cap (305). The user removes the cap (305) from the end of the body (301) and introduces the absorber (304) into the fluid, located in the rupture (310). The absorber takes up a predetermined volume of fluid, such as, but not limited to, to a final ratio of 3 parts of a reagent to 2 parts fluid. The absorber (304) serves to remove some lumps or solid particulates from the fluid, negating the need for centrifugation in a laboratory. A non-limited example use of the above apparatus may be as follows: The apparatus (300) has a cap (305) that protects an absorber (304) that is been pre-wetted with RB, located on a first arm (302). A second arm (307) carries a fluid collector (303) which includes a lysostaphin bead (306) sealed inside a rupturable chamber. The user collects the milk sample in a container. The first arm (302) of the apparatus (300) is dipped into the milk sample. The design of the fluid collector (303) allows a small volume of sample to be collected, and also allows lumps to be carried away from the main sample. Preliminary filtration tests have indicated that lumps found in mastitic milk carry similar, if not higher levels of pathogen compared to clarified milk, therefore carry-over of lumps at this stage is thought to be beneficial. Once milk is collected, the first arm (302) is urged towards a rupterer (310) which is pre-filled with a measured amount of IDTE. A temporary closure (310) is opened by a rupturing means within the rupterer (310). The amount of IDTE carried by the container is measured against the carrying capacity of the fluid collector (303) to ensure a 2 part sample to 1 part IDTE ratio. The urging motion by the user ruptures the temporary closure (e.g. by piercing a foil seal) at the bottom of the rupturer and also, simultaneously, releases a lysostaphin bead (306) for rehydrated by the IDTE, and also releases the milk sample from the fluid collector (303). The user can move the tool in a circular motion or ‘pump’ up and down the body (301) to mix the IDTE, sample and lysostaphin reagent. The rupturer (310) can now be placed inside the SHU for incubation at 37°C and 100°C, followed by the cooling down step as described in the example methods above. Once the incubation is completed, the user removes the cap (305) from the end of the second arm (307) of the apparatus (300) and dips the partially wetted absorber into the incubated sample. The absorber absorbs a set amount of sample from the rupture (310), to a final ratio of 3 parts RB to 2 part sample. This lysate is now ready for loading onto the cartridge. The absorber serves to remove some lumps from the milk sample, replacing the centrifugation step of a laboratory protocol. Apparatus comprising a captured filter Sometimes, it may be necessary for multiple filtration stages to occur, in order to remove undesirable contaminants or solid matter from a sample. For example, processing of difficult milk samples, there is the potential for blockages of microfluidics cartridges due to milk-derived particles entering the cartridge. A filter unit (400) is shown in figure 4 comprising a captured filter (401). Here, captured means that the filter (402) is held and contained within a frame. The frame may be integral to the body (403) or a separate entity entirely, as shown in figure 4. This version of the filter unit consists of two parts, the body (403), and the captured filter (401), which are not connected to each other. The captured filter (401) may contain one filter, or multiple such that multiple filtration stages may occur. This may be achieved by stacking multiple filters on top of each other. This unit (either or both of the body (403) and captured filter (401)) also uses a seal, such as, but not limited to an O-ring, to ensure a fluid-tight seal against the inner wall of the container tube. The captured filter (401) may be equipped with a a piercing piece at an end arranged adjacent a rupturable seal (404) provided on the body (403). When a user urges the body (403) towards the captured filer (401), or vice versa, such as by inserting the apparatus (400) into a container tube (405), the captured filter (401) is urged towards the body (403) upwards under pressure from the sample (406) that is inside the container tube (405), and this ruptures the rupturable seal (404) that is occluding the entrance aperture (407) of the body (403). This rupturable seal ensures no fluid (409) (such as, but not limited to, resuspension buffer) contained within a cavity (408) of the body (403) can pass into the container tube (405) unless desired by a user. Once the rupturable seal is broken, the sample will travel through the filter (402) in the captured filter (401), and into the cavity (408), and mix with the fluid (490) contained within. This action will also compress any gaseous fluid within the cavity (408). The mixed sample may then be analysed, by, for example, loading onto a microfluidics cartridge. The apparatus may be provided with a releasable cap (409) on the body (403), such as a screw-cap. The cap may be provided with a syringe plunger (410), sized complementary to a dimension (such as the diameter, if cylindrical) of the cavity (408). In this way, depression of the syringe plunger causes the volume of the cavity to be variable between maximum and minimum limits, depending on whether the piston is extended or retracted. The piston may be arranged to interfere with a stopper, such that the syringe may only be depressed a predetermined amount. Once the apparatus (400) is inserted into a sample container (405), there may be an added clamp or arresting feature which holds the apparatus in a fixed position relative to the sample container (405) in order to prevent the apparatus moving due to the built-up pressure in the cavity (408). Following filtration, the user would remove the body (403) from the container 405, and disassociate the captured filter 401 from the body. The body may be connected (such as via complementary screw threads) to a microfluidics cartridge, and the cavity (408) contents (409) injected into the microfluidic system by depressing the syringe (410). Such actuation of the syringe (410) may either be manually, by user action or automated via a suitable device or system. If a one-stage filter 402 is used in the captured filter (401), then a secondary filter may be placed at a suitable location within the body (403), or indeed a separate accessory filter may be provided to be used with the apparatus (400), such as on the microfluidics cartridge itself.. The syringe part (410) may be hidden from the user, via a suitable cover or access point. The plunger may be arranged to be completely housed, and not protrude from, the releasable cap of the apparatus (400). This way, accidental actuation of the plunger can be prevented, and only a suitable system or trained technician will be able to actuate the plunger and expel sample for analysis. A non-limited example use of the above apparatus may be as follows: A milk sample is collected in a generic collection vessel with ideal volume between 20 and 100 mL, to allow for easier handling. The diameter of the vessel may be relatively large (such as approximately 5-10 cm), to allow for easier sample collection directly from a lactating animal or parlor line. The container may be provided with a marker, such as a ‘fill-to-line’, to ensure users do not overfill this container. The volume collected at this stage is not essential, as long as it is sufficient to allow for a sub-sample to be taken. A small sub-sample is aspirated out of the generic collection vessel, such as by using a 1 mL syringe. A larger capacity syringe could be used, however this will reduce accuracy. Once the milk sample is aspirated into the syringe the sample is mixed with a lysostaphin bead. The lysostaphin bead will be rehydrated and the enzyme will dissolve into the milk sample. The volume of the sample collected here is 400 pL. However this volume can be scaled in line with the volumes of other reagents, to better match the user case and / or cost requirements of the product. The user then dispenses the whole volume (400 uL) into a container (405) (a suitable tube such as a 2 mL tube) which is pre-filled with 200 pL of IDTE (1X TE buffer, pH 8.0). The action of dispensing the milk sample into the IDTE allows for some mixing of sample, IDTE and lysostaphin to occur, however it is advisable for the user to invert the container (405) a few times to ensure reagents are mixed adequately. The user places the container (405) into a sample handling unit (SHU); this is a heating and cooling dry bath unit which is pre-programmed to incubate the sample at 37°C for 10 minutes, followed by heating at 100°C for 15 minutes. The SHU will then cool down the sample to an acceptable temperature close to room temperature (20°C). The SHU takes some time to ramp up and down during incubation, and actual incubation times at the given temperatures can be further adjusted (primarily reduced) to account for the time spent ramping, in order to reduce total processing time. At this stage, the user will fit the body (403) and captured filter (401) into the container (405) and apply pressure in order to simultaneously filter the incubated sample and mix the sample with rehydration buffer (409, RB), which is already pre-loaded into the body (403). The body (403) is of exact size which allows a tight seal between the inside of the container (405) and the outside of the body (403); this is further assisted by an O-ring which is placed on the body (403). The body (403) also includes a filter (402), which removes lumps and debris from the filtrate, and a rupturable seal which prevents the pre-loaded RB from leaking out of the body (405) during storage and handling. The volume of RB is measured to be 225 pL, in order to achieve a ratio of 2 part sample to 3 parts RB following filtration. The user then removes the body (403) from the container (405) and may connect it with an analysis cartridge. The lysate is then loaded onto an analysis cartridge by depressing the syringe piston (410).

Claims

1. An apparatus, suitable for preparing a sample for analysis, the apparatus comprising:a body defining a cavity with an entrance aperture;a seal arranged on the body to, in use, engage with a container and form a liquid tight seal 5 between the container and the body;a valve arranged in fluid communication with a filter and configured to prevent fluid contained within the body from passing through the entrance aperture;wherein the filter is arranged in fluid communication with the entrance aperture; andwherein the cavity comprises a rehydration buffer solution.10 2. The apparatus of claim 1, wherein the apparatus includes, or forms part of, a filter unit.

3. The apparatus of claim 1 or 2, wherein the seal is an O-ring and / or wherein the valve is an umbrella valve.

4. The apparatus of any one of claims 1 to 3, wherein the valve is arranged adjacent the filter.

5. The apparatus of any one of claims 1 to 4, wherein the fluid is a sample.15 6. The apparatus of any one of claims 1 to 5, wherein the filter is arranged adjacent theentrance aperture.

7. The apparatus of any one of claims 1 to 6, wherein the body comprises an egress aperture arranged in in fluid communication to the entrance aperture.

8. The apparatus of claim 7, wherein the egress aperture is arranged in opposition to the 20 entrance aperture.

9. The apparatus of claim 7 or claim 8, wherein the egress aperture is provided with a temporary closure.

10. The apparatus of claim 9, wherein the temporary closure is a film or foil.

11. The apparatus of claim 9 or claim 10, wherein the temporary closure is peelable.25 12. The apparatus of any one of claims 1 to 11, wherein at least a portion of the body isdimensioned complementary to an internal dimension of a container, such that insertion of the body into the container forces a fluid contained within the container through the filter, valve, and into the cavity.

13. The apparatus of claim 12, wherein the apparatus further comprises a container and the container is a sample processing container.

14. The apparatus as claimed in claim 12, wherein the apparatus further comprises a container, and the container comprises a fluid absorber, the fluid absorber arranged to absorb 5 fluid and to then release the fluid upon insertion of the body into the container.

15. The apparatus of claim 14, wherein the absorber, in use, provides a filter.

16. The apparatus claim 14 or 15, wherein the absorber is pre-treated with a reagent.

17. A kit of parts comprising an apparatus as claimed in any one of claims 1 to 16 and at least one container.10 18. The kit of claim 17, further comprising one or more from the list: a fluid, a liquid, and / or acap.

19. A kit of parts as claimed in claim 18, when comprising a cap, wherein the cap is configured to occlude the entrance aperture.

20. A kit of parts for collecting, filtering, and preparing a sample for analysis, the kit comprising:15 a container;an apparatus as claimed in any one of claims 1-16; anda processing container.

21. A kit of parts as claimed in claim 20, wherein the kit further includes a heater and / or an analyser.20 22. The kit of parts as claimed in claim 21, wherein the analyser comprises a further filter.

23. The kit of parts as claimed in any one of claims 20 to 22, wherein the container comprises an aperture sized to allow collection of 20- 100 ml of a fluid.

24. The kit of parts as claimed in claim 23, wherein the container comprises an aperture sized to allow collection directly from a living creature or processing line.25 25. The kit of parts as claimed in claim 24, wherein the container comprises an indicatormarker.

26. A method of preparing a sample for analysis, using a kit as claimed in any one of claims 20 to 25, the method comprising:collecting a sample in the container;treating the sample with a reagent;adding a buffer solution to the sample;heating the sample to a first temperature for a first period of time, heating the sample to second temperature for a second period of time, and cooling the sample to ambient5 temperature; andhandling the sample with the apparatus.

27. The method of claim 26, wherein the below steps are performed in the following order:(a) collecting a sample in the sample container;(b) treating the sample with a reagent;10 (c) adding a buffer solution to the sample;(d) heating the sample for a first period of time, heating the sample fora second period of time, and cooling the sample to ambient temperature.

28. The method of claim 26, wherein the method further comprises the step of analyzing a filtered sample.15 29. The method of any one of claims 26 to 28, wherein the reagent comprises a lysing agent.

30. The method of claim 29, wherein the reagent comprises a bacteriocin.

31. The method of claim 30, wherein the bacteriocin is lysostaphin.

32. The method of any one of claims 26 to 31, wherein the first temperature is 37°C and the second temperature is 100°C.Application No: GB2401017.5Examiner:Contract Unit ExaminerClaims searched: 1-17 &61-68Date of search: 26 November 2024Patents Act 1977: Search Report under Section 17Documents considered to be relevant:Category Relevant to claims Identity of document and passage or figure of particular relevance X 1-8, 10- 17, 61-63 US2011 / 233119 Al (NELSON STEVEN D) paragraphs [0055], [0056]; figures 25, 26, 11, 12 A - US9677981 B2 (INNOVATIVE PROPERTIES CO) column 1, lines 29-34; figures 6A-C A - US11045036 B2 (O2C RALEIGH LLC) figure 13 A - US3512940A (SHAPIRO JUSTIN J) figures 1-3 A - US8211384 B2 (ELLIS SAMUEL A; HINGORANI KISHAN GET AL) figures 3, 4 A - US4510058A (CAIS MICHAEL ET AL) figures 1-3Categories:X Document indicating lack of novelty or inventive step A Document indicating technological background and / or state of the art. Y Document indicating lack of inventive step if P Document published on or after the declared priority date but combined with one or more other documents of same category. before the filing date of this invention. & Member of the same patent family E Patent document published on or after, but with priority date earlier than, the filing date of this application.Field of Search:Search of GB, EP, WO &US patent documents classified in the following areas of the UKCX :Worldwide search of patent documents classified in the following areas of the IPC_____________BOID; BOIL_________________________________________________The following online and other databases have been used in the preparation of this search reportInternational Classification:Subclass Subgroup Valid From GOIN 0001 / 10 01 / 01 / 2006 BOID 0033 / 01 01 / 01 / 2006 BOIL 0003 / 00 01 / 01 / 2006

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