GB2512141A

GB2512141A - Encapsulation System

Info

Publication number: GB2512141A
Application number: GB201305329A
Authority: GB
Inventors: Graham Scott Gutsell; Stuart Charles Wright
Original assignee: Individual
Current assignee: Individual
Priority date: 2013-03-22
Filing date: 2013-03-22
Publication date: 2014-09-24
Also published as: GB201305329D0

Abstract

A device 1 comprising a protective enclosure 6, a first module 2 and a second module 5 wherein the modules are attached to the protective enclosure such that the protective enclosure lies between the modules and the protective enclosure is punctured, perforated or pierced to facilitate flow of reagents between the two modules. Ideally the first module comprises moisture sensitive reagents and the second module comprises wet reagents. The protective cover is preferably formed of laminated aluminium material. The first and second modules can comprise conduits, reservoirs, channels, etc which are aligned with corresponding components in the other module. The first module may comprise a sensor or fluid manipulation platform. The first module may also comprise discrete pots (12;fig 4) for storing moisture sensitive or dry reagents which are connected to a fluid conduit (7;fig 4) by spigots (13;fig4). The device is ideally used in detection or diagnostic tests where reagents existing in one or more physical phases can be stored together.

Description

ENCAPSULATION SYSTEM

The present invention relates to systems used in the detection or diagnosis of certain target materials where one or more of the reagents employed in the test is sensitive to the presence of liquids, dampness or moisture, but where one or more additional reagents are themselves either liquid, damp or moisture-containing. The former will be referred to here as either dry, or moisture-sensitive, reagents and are likely to be biological materials prepared for use by freeze-drying (or lyophilisation); the latter will be referred to as either wet, or liquid, reagents. Such wet reagents may be, for example, a simple buffer, diluent or solvent. As a consequence of their moisture-sensitivity, and in order to provide stability during prolonged periods in storage, the dry reagents frequently have to be packaged in a protective enclosure, such as a pouch, bag.

sachet or cassette, the walls or sides of which are resistant to the transmission of moisture.

Laminated aluminium foil pouches are commonly used for this purpose.

Where they are already in use, such tests are typically conducted on or within a disposable element, device or assembly, which may be stored inside the same protective enclosure as the dry reagents and to which the additional wet reagents are added at or near the time of use.

Commonly, the dry reagents are stored on or within the test device. To perform the test, an operator needs to remove the device and any dry reagents from its protective packaging.

Furthermore, such tests are frequently operated by automated instrumentation which, in addition to controlling the sequence and timing of the test, may also provide additional energy input and be involved "reading" the output signal from the test reaction.

A frequent shortcoming of such tests is that the wet reagents cannot be kept, ready for use on the test device and inside the same protective packaging, without causing some deterioration to the dry regents during long-term storage. Thus to add, mix or otherwise combine any wet and dry reagents together in the appropriate manner and at the appropriate time(s), there is a requirement for additional steps to be performed, either by an operator or by further automation.

A solution to this shortcoming would be desirable simply for convenience, however, it would be especially important for any tests that need to be deployed, either; a) with minimal user-intervention, for example in very busy settings where speed may be critical or in situations where users may have limited training; or b) in semi or fully-automated applications where users are not necessarily present, such as at-line tests conducted within process plant and machinery.

The present invention overcomes this shortcoming by storing a first modular part of the test device together with the dry reagents in' -_ nclosure, such as a welded pouch, but affixing a second modular part (or parts) of the device together with the wet reagents to the outside of the enclosure in such a way that they can be dispensed when require through one or more holes formed in the enclosure wall and delivered to the required position(s) on or around the device. The protective enclosure thus effectively becomes a functional part of the device. By such means, the wet and dry reagents can be stored for prolonged periods in a unified assembly, which is amenable either, to very simple manual operation or to automated operation.

One example, described in this application, is a test for the detection of endotoxins, such as that used during the manufacture of injectable pharmaceuticals or certain medical devices. The invention thus relates to a disposable cartridge containing wet and dry reagents into which a test sample is loaded, and that in combination with a sensor or detector, permits the presence of endotoxins in the sample to be detected at relatively high speed and low cost. The example described is suitable for use at-line within process plant for pharmaceutical liquids, such as water for injection (WFI).

Further instances of tests involving both wet and dry reagents and which need to be performed with the minimum of user-intervention, can be found in the field of human diagnostics.

Additionally, with the introduction of tests that are conducted at the point-of-care (POC tests), ease-of-use is a particularly important requirement and indeed, there may be regulatory requirements that mandate this. A second example described in this application is a POC molecular diagnostic test.

The prior art contains many examples of devices used in the various fields of detection and diagnostics and that make use of both wet and dry reagents. Two such cases, that provide relevant background to the examples of the present invention described hereinafter, are as follows.

The use of Limulus amebocyte /ysate (LAL) assays have become the standard test for the detection of endotoxins (or lipopolysaccharides). Endotoxins are liberated when the outer cell membrane of gram-negative bacteria are destroyed, and in humans, can be responsible for severe toxic shock or sepsis, which frequently can be fatal. Commonly, tests involving LAL are either gel-clot, turbidimetric or chromogenic assays, and currently, are normally conducted in a laboratory remote from the point of sampling. These tests typically suffer from the complexity of the associated test protocols, time delay between sample collection and actual testing, and the requirement for skilled labour. Furthermore, in the case of WFI testing, considerable volumes of water can be consumed by production processes before endotoxin levels have been determined, since testing is not normally conducted at-line. W02005/010207 discloses a cartridge-based system in which disposable cartridges are pre-loaded with one or more imrriobilised hemocyte lysates. In use, a sample is admitted to the cartridge via an inlet port and is then moved along a conduit where it reconstitutes and/or solubilises a dried LAL reagent that is disposed on a fluid contacting surface of the conduit before being moved to an sensing cell to permit observation of a change in optical properties. The test is normally performed by an operator who can load the additional necessary wet reagents onto the device at the time of use.

However, there are applications where the device could be performed away from the laboratory and for which it would be advantageous for all the reagents to be stored on-board a test device, which can be fed into a small and relatively simple controller, potentially without the need for manual intervention. These applications would thus benefit from the present invention.

W0201 1051735 describes a device that can perform a biochemical reaction such as a DNA amplification reaction. The device thus has potential applications such as detecting the presence of infectious diseases, in either animals, or indeed humans. When deployed in this application, the device may contain dry reagents in one or more of the wells cited, and to provide it with long-term storage stability, may be packaged in a moisture-resistant pouch.

However, in addition to adding a test sample at the point of use, various liquid reagents need to be loaded onto the device to enable its correct function. One of these would be added along with the sample, a second takes the form of a diluent that is mixed with the products of DNA amplification before analysis on the lateral flow strip described. This device thus requires certain manual steps, but could benefit from the present invention whereby the entire process could be performed without manual intervention after the sample has been introduced.

The present invention has some similarities with the prior art devices cited above, but seeks to provide multiple novel improvements over these and other existing solutions.

In one aspect of the present invention, a first part, module or sub-assembly of the test device (hereinafter referred to as the first module) is encapsulated by being attached to the inside surface of a protective enclosure, such as a pouch, bag, sachet or cassette. For ease of any forming and joining, the enclosure can be manufactured from a ductile metal, such as aluminum, which has been laminated with a thin polymeric material. A laminated aluminium sheet or foil may be advantageous for this purpose. A second part, module or sub-assembly of the test device (hereinafter referred to as the second module) is attached to the outside surface of the same protective enclosure. The first module contains, or has associated with it in some other way, the dry reagents that are employed by the test; the second module contains, or has associated with it in some other way, the wet reagents that are employed in the test.

Furthermore, the two modules are attached to opposing sides of a wall (or walls) of the protective enclosure such that certain wells, reservoirs, conduits, channels, ducts, vias, pipes, wicking materials or other fluid-containing or conveying features of the first module are aligned with corresponding wells, reservoirs, conduits, channels, ducts, vias, pipes, wicking materials or other fluid-containing or conveying features of the second module. In operation, holes may be formed in the protective enclosure by puncturing, perforating or other hole-making means such that the wet reagents from the second module may pass into the first module in order to be combined with dry reagents located therewith and/or otherwise deployed.

The two modules may be attached to their respective sides of the protective enclosure wall(s) by joining-means such as sealing, welding or bonding. This may be further enhanced by additional adhesive materials such as single-or double-sided adhesive tapes, similar to those used in many disposable diagnostic devices.

By virtue of the two modules being attached to opposite surfaces of the same region of the protective enclosure wall(s), their positions are accurately defined with respect to each other.

Thus the position of the first module, or of specific features within it, are known provided that the position of the second module is at some specified location. This may be, for example, as a consequence of it being retained in some piece of automated instrumentation or controlling equipment (hereinafter referred to as the "controller"). The benefits of this should be made apparent by what follows.

In addition to forming holes in the protective enclosure for the transport of liquids between the two modules once the controller has located onto the second module, it can then, if necessary! form additional openings, apertures or windows in the enclosure at specific locations in order to gain access to certain functional areas, such as other fluid ports, pneumatic or vacuum coupling ports, venting ports, to make electrical connections, to perform sensing operations or to transfer modular elements (such as special reservoirs or transducers) from one module to the other. In other applications of the invention the area of the protective enclosure that does not form the connecting membrane between the first and second modules may be partially or completely removed, either by an operator prior to the start of the test or by the controller.

It should be noted, that in contrast to the need to form apertures in the membrane for the functions described above, some applications of the invention, especially those involving biochemical reactions! require the transfer of heat to specific regions of the device. It may be necessary, for example! to perform an incubation, amplification or culturing step. Clearly heat can be transferred through the enclosure wall or membrane to specific regions of the first module purely by means of thermal conduction without the need to form an aperture. This is perceived by the inventors as a particularly beneficial feature associated with the invention.

Furthermore, it is advantageous for the second module to contain registration features by which any controller can locate and retain it whilst performing the test. These may simply be the outer edges of the second module, but beneficially are specific registration features, such as holes into which locating pins can engage. There is no need, therefore, for the first module to contain similar registration features, since it has been located and retained by virtue of its attachment, through the enclosure membrane, to the second module. [Notwithstanding this, it may be advantageous for the first module to contain different registration features that are used during manufacture, as described later in this document.] In order to form perforations in the membrane wall of the enclosure to convey the wet reagents through it and between the modules or for other functions to be performed, either module can contain suitable elements, such as cutting pins, pegs, blades, punches or other protruding features. It is preferable for these to be actuated by mechanisms within the controller, and so these are preferably contained in the second module. Alternatively, any protruding features can be located in the first module and puncture the membrane through the displacement of the same under the action of differential pressure, for example. The puncturing elements can be located inside the reservoirs or conduits of the second module that contain or convey the wet reagents provided that some part of the construction is deformable so as to allow movement.

The puncturing elements can be made from specially hard materials, however, if the protective enclosure is relatively thin-walled, then these features can be made from softer materials such as injection-moulded plastics. This is clearly advantageous to enable the mass manufacture of component parts.

The puncturing elements can be of various geometries to achieve different purposes, for

example:

-to create a small punctured hole through which a liquid can be conveyed from say a reservoir in the second module to a channel in the first module, the puncturing element may take the form of a small cylindrical pin.

-a similar geometry may be used for pneumatic or vacuum couplings, or for air vents.

-to remove a larger portion of the membrane so as to create an aperture to allow greater flow between wells in both modules for example, the puncturing element may be a punch. The end profile of the punch may take various different forms depending on the necessary aperture to be created, for example; it may be square-ended, angled (like a hypodermic syringe), concave (like a stationary hole punch), conical (like a drill point), or a cruciform shape. A unified well straddling the two modules may be created this way, if for example freeze dried reagents retained in a well in the first module need to be combined and re-suspended by liquid from an aligned reservoir in the second module.

-in some situations it is also beneficial for a small element (which in storage resides in the second module) to punch' a hole in the membrane (as above) and then be transferred completely into the first module. The element may contain a special well or a sensing transducer, for example, but may not be suitable for long-term storage in the first module with the wet reagents.

In a manner that is not dissimilar to the above, instead of the second module containing puncturing elements, it can contain pressing elements so that the controller can exert localised forces through the membrane (thereby not puncturing it) to the first module. Alternatively, the pressing elements can form part of the controller and thus exert forces in a similar fashion through the membrane. These pressing elements can perform various functions at different times as required by a particular process, for example -to obstruct channels within the first module and thus effectively perform the function of valves to restrict or prevent the motion of fluids in the first module -to squeeze wicking materials, either to modify their capillary action or to completely inhibit flow along them -to bring wicking materials into contact with each other in order to initiate fluid transport -to initiate fluid movement through the creation of differential pressure -to cause electrical connections to be made.

In a further adaption of similar principles, if the protective enclosure forms a partition between two chambers, one in each module, there can be communication of pressure without the flow of any fluids. This can be used to create a simple pressure transducer, for example.

A further important aspect of the invention is the manufacturing method by which the device is fabricated and assembled.

The protective enclosure can be created in various different ways. It can be formed from a single laminated sheet or foil of material that is wrapped around the first module prior to welding along its edges. Similarly the enclosure can also consist of multiple pieces. However, for ease of mechanical handling it has been found that it is preferable for the enclosure to be formed from two separate pieces. In applications where the first module is generally flat in shape, resembling a credit card for example, the two pieces can both be foil materials that forms a pouch around the first module. If however, the first module is deeper, it is generally preferable for one of the two pieces to be mechanically pre-formed to create a tray or cup shape that encloses its volume, and for the second piece to be a foil that is attached thereto. This arrangement bears similarity to the packaging used on some coffee capsules where the grounds are stored in a protective environment up to the time of use.

Beneficially, certain features on the first and second modules can be present to improve the mechanical handling of the component parts during the production processes.

Whilst the inventors have identified both rotary and linear analogues of the assembly process, the key aspect of the corresponding machinery is that it has two module retention mechanisms (described in the following description as the first and second retention mechanisms), one to hold each module respectively, and that they interleave or overlap each other in such a way to provide a "handover' and thereby, a means to form the protective enclosure by bonding or welding together one, two or more sheets of protective enclosure membrane material.

A rotary version of the process utilising two indexing carousels that overlap at one angular position has generally been found to be preferable since it can simplify the mechanical handling requirements and increase manufacturing throughput. However, in some applications it is advantageous to link multiple test devices together in a chain-link structure (for automated feeding by the controller, for example), a semi-continuous linear analogue of the system, comprising two reciprocating gripper arms, provides a very suitable alternative.

The sequence of either linear or rotary processes may proceed as follows: 1. The dry reagents are loaded into the first module, (Note that the first module may contain sub-component parts specifically arranged to store the dry reagents, possibly to simplify such pre-processing as lyophilisation. If so these would preferably be added to the first module at or before this step 1).

2. The first module is loaded onto the first retention mechanism being located by certain registration features that allow its position to be accurately defined relative thereto, albeit to a moving frame of reference.

3. The first of the two sheets of protective membrane material is affixed to the first module, preferably to a generally flat region thereof. Joining means, including sealing, welding or bonding, may be suitable for this operation.

4. In parallel with any of the preceding steps, the second module, or a sub-component part of it, is loaded onto the second retention mechanism and located by certain features that allow its position to be accurately defined relative thereto.

5. Whilst the first module is retained in the first retention mechanism, the second module, or a sub-component part of it (such as a refeience frame), is positioned relative to the first module with the desired final alignment and is then affixed to the opposite surface of the first sheet in a defined position. Joining means, including sealing, welding or bonding, may be suitable for this operation.

6. The first module is then released from the first retention mechanism, the welded or bonded assembly now being retained by the second retention mechanism.

7. The second sheet of protective membrane material is then bonded or welded to the first sheet, in such a way that the first module is enclosed between the two.

(Note that steps 1-7 may be performed in a controlled dry environment to protect the dry reagents).

8. Any remaining sub-component parts of the second module can then be attached to the initial sub-component part attached to the first sheet. Any wet reagents may have been pre-loaded into the respective reservoirs in the second module, or can be loaded as an integral process within this step 8.

9. Alternatively and finally, the completed second module can be loaded with the wet reagents.

As described above, two examples of the invention are presented here, each building on the prior art cited above. The second of the two examples describes in greater detail the manufacturing method which is clearly applicable to either.

In the first example, the device (1) incorporates a first module (2) pre-loaded with dried reagents for use in hemocyte lysate based assays as shown in Figure 1. The first module is separated from one or more external reservoirs (3, 4) integrated into the second module (5) by the wall of a protective enclosure (6) which may or may not be in the form of a pouch entirely surrounding the first module. Figure 2 (protective enclosure 6 not shown) illustrates how the first module incorporates one or more conduits (7) permitting the transport of fluids, and may include wells (8) to enhance the mixing of samples and reagents as desired. One or more sensing cells (9) is included to permit observation of a change in a material property. Such sensing may be optical (such as turbidity, colour change, change in absorbance or transmittance or change in fluorescence), or may detect changes in other physical properties, such as viscosity.

Additionally, one or more vents (10) may be provided. One or more of the external reservoirs contains high purity water (HPW) or an alternative neutral hydrating agent, and at least one of the external reservoirs contains a gaseous fluid such as air.

The dried reagents may include one or more of lyophilised LAL, one or more standard endotoxins, one or more chromogenic substrates and other performance enhancing agents such as stabilisers. Such reagents are pre-loaded at various locations along the conduits such that, depending on the specific use, the LAL reagent is hydrated by the water (or other hydrating medium) or by the test sample entering the device along the one or more conduits (7). In the case of a chromogenic test the sample (or a mixture of sample and reagent) additionally contacts a chromogenic substrate before entering the sensing cell. The chromogenic substrate may be either upstream or downstream of the [AL. In the case of a standard or positive control test, the water (or other hydrating medium) hydrates a lyophilised endotoxin of known concentration before it reaches the [AL reagent.

The external reservoir assembly shown in cross-section in Figure 3 forming the second module (5) or part thereof and the portion of the first (internal) module (2) to which it is adjacent are preferably designed to incorporate a pin (11) or other sharp feature which can be moved to perforate the protective enclosure (6) thereby providing direct communication between the reservoir(s) and one or more conduits (7) in the first module, and hence permitting the transport of the reservoir contents into one or more of the conduits in the first module. In this way, water or other fluid stored in a reservoir outside the protective enclsoure can be admitted to reconstitute or solubilise dry reagents inside the internal module at the time of use. Puncturing of the membrane may alternatively be achieved through a sharp feature in the first module adjacent to the reservoir over which the protective enclosure membrane material is strained by the generation of a relative positive pressure in the liquid reservoir.

The design of the individual reservoirs in the first module may advantageously be such that the reservoir can be squeezed, deformed or depressed to generate a relative positive pressure within the reservoir such that fluids stored in the reservoir are transported in a controlled way into and along conduits within the first module. Such reservoir pressurisation may also be instrumental in perforating the membrane as above. Reservoirs of a blister' form may be advantageous.

According to another aspect of the present invention, the dry reagents are accommodated in discrete pots (12) shown in Figure 4 which, at the time of cartridge assembly, are admitted and sealed into location points or mounted onto spigots (13) in the first module (2) such that there is fluid communication between the reagent and one or more conduits (7) in the first module.

Advantageously, the dimensions of the location point or spigot are such that the pot is a significant interference fit into the location point or onto the spigot respectively, thereby achieving the required sealing and security. The size of the pots is small relative to the internal module and hence the packing efficiency in the drying cycle of any lyophilsation process is much greater than with existing cartridge solutions, with concomitant reduction in costs of drying.

The principle of operation of the cartridge is described with reference to Figure 5 (protective enclosure membrane 6 not shown) which shows a simplified version of a test cartridge, in this instance for constructing a standard response curve whereby a control standard endotoxin (CSE) is used to generate a response from the [AL reagent. Two separate conduits (14,15) transport high purity water (or other hydrating medium) from inlet ports (16, 17) ultimately to a common conduit (20) along which is located one or more sensing cells (9) and, depending on application, a chromogenic substrate (21). Lyophilised LAL (18) is stored in a pot (12, not shown) along conduit (14) and lyophilized CSE is stored in a pot (12, not shown) along conduit (15). Water or other fluid is admitted via the inlet ports (16,17) prior to which the protective enclosure membrane (6, not shown) is perforated by one of the methods described above as well as optionally, at one or more vents (10). A controlled volume of water solubilises/reconstitutes the lyophilised LAL and CSE materials respectively whereupon they are pumped to the common conduit (120). A defined volume of both materials is driven along the common conduit (20) and/or mixing wells (8) provided for this purpose. Following an appropriate incubation period the composite mixture of sample and reagent is transported to the sensing cell (9) to permit observation or detection.

For compliance with recognised quality standards it is necessary to perform positive product and negative control tests and to construct a standard response curve for each sample under test. These are all performed at least in duplicate. Comparing with the process described above: -for a negative control test, the CSE material is omitted -for a product positive control (which checks for cross-reactivity between the sample and CSE), the test is identical in principle to that shown above except that sample material is admitted in inlet port (16) in place of pure water -for an actual sample test, sample material is admitted into inlet port (16) in place of water and the CSE material is again omitted.

From the description above it will be apparent that some or all of these individual tests can be combined on a single (albeit more complex) test device. In these instances, an individual first (internal) module may have a multiplicity of conduit pathway sets but these may be so arranged that multiple pathway sets are served by common external reservoirs and sample inlet port. A multiplicity of pathway sets permits a sample to be tested (multiple times) and control tests to be done on a single cartridge.

It will be further appreciated that in place of the sensing cell (9) there may be a discrete sensor or transducer that detects a response to endotoxin contamination by means of other physical phenomena integrated into the first module. Such a sensor may require electrical connection to be made to the controller. Alternatively, the first module may incorporate an additional fluidic manipulation platform (22) that can perform the detailed fluid transport and mixing steps necessary to perform all the individual tests (sample, controls and standard curve) necessary in the entire assay, once the solubilisation of the freeze dried materials stored in pots (18,19) has been completed by water and test sample admitted at points (16,17) by means of the steps described above. The sensing cell(s) (9) can be within the fluidic manipulation platform. This integrated cartridge incorporating the fluidic manipulation platform is illustrated in Figure 6.

Suitable materials for the first module include metals, alloys, glasses, ceramics, polymers. It is anticipated that the first module will be essentially planar in form with the conduits positioned within the body or at the surface of the module. Advantageously, the first module may be composed of elements including multiple planar sections in at least one of which are formed the conduits and at least one of which are formed the ports and or wells, such elements being joined together. In one aspect of the invention, the internal module may comprise two such planar elements of proportions such that the resulting internal module is also predominantly planar in form.

The manufacturing processes adopted should preferably ensure that all material surfaces which could contact the sample or the reagents in use are free of the microbial contaminant, especially endotoxins, that the cartridge is intended to be used to test.

While manual operation and observation of the simple single test cartridges is possible, it is intended that the cartridge be used in an automated reader instrument in which temperatures, timings and fluid motions are accurately controlled and the optical observation accurately monitored and recorded. Pneumatic and/or electromechanical activators in the instrument operate on the external reservoirs on the cartridge or otherwise to effect the puncturing of the protective enclosure membrane and control pumping of the fluids within the cartridge, and admit a controlled volume of the sample (or LAL in place of sample for a negative control assay).

The fluid samples and hydrated reagents may advantageously be transported back and forth within the conduits and/or wells to optimise mixing. To simplify the cartridge, especially in cases where there is a multiplicity of conduits, there may optionally be one or more valves in the instrument optionally connected to, or in fluid communication with, the vents (10) in the internal module such that individual conduits may be subjected to pressure differentials to move fluid contents therein.

Optionally, such instrument could be plumbed into a Water For Injection (WFI) system or other process plant such that sampling, monitoring and recording is undertaken automatically with results transmitted, optionally wirelessly, to a central laboratory or quality control repositoly. In this situation, it is advantageous to have the test devices manufactured as a continuous-feed string so that they can be fed through the controller as required. As described above, the linear variant of the manufacturing process may be preferable for manufacturing the devices in this format.

The second example is a further development of the ideas disclosed in W0201 1051735 cited above, such that the device disclosed in the prior art essentially forms just the first module. This arrangement could enable a simple POC molecular diagnostic test. The device or first module (2) illustrated in Figure 7 contains a well (24) in which a biochemical reaction takes place; a diluent channel (26) leading into the well and; an analysis channel (27) leading from the well to a lateral flow device (28). The device (2) is enclosed in a protective enclosure (6). Attached to the outer surface of the pouch is a second module (30) incorporating a diluent reservoir that connects to the distal end of the diluent channels (26) through a hole punctured in the protective enclosure membrane (6). The working details of the prior art device that forms the first module (2) are extensively described in the prior art documents and so are not repeated here.

The diluent reservoir contains an integral pin capable of puncturing the protective enclosure membrane (6) when the contents need to be admitted to the reaction. Furthermore these can be actuated, either by manual or automated means.

A brief description of the rotary variant of the manufacturing process is provided here. In addition to its own specific benefits, this should help to further illustrate the key features of the invention.

The first module (2) is loaded at position (31) onto a first retention mechanism (32) within indexing carousel (33); this is advanced into an assembly/welding station (34). The first of the two sheets of protective enclosure membrane material (35), which here are laminated foil of aluminium and a polymer, such as polyethylene, is fed over the top of the conveyor from a reel (36). At the first assembly/welding station (34), the carousel is supported and a welding head (37) containing a heated platten descends and maintains a constant force over the foil pressing it onto the first module for a defined but relatively short period of time, thereby effecting a welded bond between the polyethylene layer and the plastic material from which the first module is made. The welding head (37) also contains a die-cutting blade that crops the assembly from the web of laminated material (35). In a second assembly/welding station (38), the second module (5) is positioned over the initial assembly and joined to the upper face of the foil material. This may also be a welded joint, however for clarity, the welding head not shown in the diagram. This assembly is then gripped and retained by the second retention mechanism (39) which forms part of the second carousel (40). Before the carousels indexes again, the first retention mechanism (32) on the first carousel (33) releases. The assembly then proceeds around the second carousel (40) to a third assembly/welding station (41) at which the second sheet of protective enclosure membrane material (42) that is fed from a reel (43) and welded to the underside of the assembly so as to completely encapsulate the first module. The welding head (again not shown in diagram for clarity) also contains a die-cutting blade that crops the laminate material in order to separate it from the web on reel (43). At the final position (44) on the second carousel, the assembly can either be released into a collection bin, or passed to a downstream handling system for subsequent assembly finishing steps.

In a further development of this example of the present invention, the second module contains a sample preparation sub-system that automates the extraction of genomic DNA from an otherwise raw" or unprepared sample. Some variants of this sub-system contain a very simple denaturing step to release DNA, but in more sophisticated variants offering higher yield and sensitivity, the sub-system might utilise a functionalised solid-phase extraction material such as a porous membrane or magnetic beads, and operate a sequence based around the following: -lysis to release DNA from cells (such as for example blood or buccal cells) -binding of this DNA to the functionalised solid-phase material -removal of cellular debris by multiple cleaning/washing steps -elution of the DNA from the solid-phase material -finally transfer of the eluted DNA into the first module for amplification, by either a thermal-cycling or isothermal process.

When more complex sub-systems such as the one described above are used, it is advantageous to attach this after assembling the core test device. Thus, instead of joining the entire second module to the protective enclosure membrane material as a single step, it is more common to attach a sub-component that acts as a reference frame (see step 5 in the manufacturing sequence above) to which the remaining components can be added by a subsequent process.

Descriptions of the drawings

Figure 1 -a schematic drawing illustrating the separation of the internal module from the external reservoirs by the protective enclosure.

Figure 2-a schematic illustration of an internal module wherein two separate conduits transport high purity water (or other hydrating medium) to re-suspend lyophilised reagents, ultimately to a common conduit in which is located one oi more sensing cells.

Figure 3-a schematic illustration of an external reservoir and puncturing mechanism.

Figure 4-a schematic illustration of a reagent containing pots and a spigot onto which it connects.

Figure 5-a schematic illustration of an internal module by reference to which the principle of operation is described.

Figure 6-a schematic illustration of the system where a fluidic manipulation platform is integrated.

Figure 7-a schematic illustration of a simple POC molecular diagnostic test.

Figure 8 -a schematic illustration of a rotary variant of a suitable manufacturing process.

Claims

CLAIMS1. A device comprising a protective enclosure and first and second parts, modules or sub-assemblies attached to said protective enclosure to facilitate the combination and/or deployment of reagents stored therein.
2. A device according to claim 1 where the first part, module or sub-assembly is attached to the inner surface of the protective enclosure and the second part, module or sub-assembly is attached to the outer surface of the protective enclosure.
3. A device according to either claim 1 or 2 where the first part, module or sub-assembly contains, or has associated with it, certain moisture sensitive reagents and the second part, module or sub-assembly contains or has associated with it certain wet reagents.
4. A device according to claim 3 where said moisture-sensitive reagents are freeze-dried reagents.
5. A device according to any of the above claims where the first and second parts, modules or sub-assemblies are attached to opposing sides of a wall or membrane of the protective enclosure.
6. A device according to any of the above claims where the protective enclosure is partially or fully made from a laminated aluminium material.
7. A device according to claim 6 where a part of the protective enclosure membrane is a cold-formed component.
8. A device according to any of the above claims where the first and second parts, modules or sub-assemblies are attached to opposing sides of a wall or membrane of the protective enclosure such that certain wells, reservoirs, conduits, channels, ducts, vias, pipes, wicking materials or other fluid-containing or conveying features of the first module are aligned with corresponding wells, reservoirs, conduits, channels, ducts, vias, pipes, wicking materials or other fluid-containing or conveying features of the second module.
9. A device according to any of the above claims that further comprises puncturing, perforating or other hole-making means.
10. A device according to any of the above claims where the first part, module or sub-assembly further comprises a discrete sensing element or fluid manipulation platform.
11. A device according to any of the above claims where the first and second parts, modules or sub-assemblies are attached to the protective enclosure by sealing, welding, bonding or by the addition of adhesive tape.
12. A device according to any of the above claims where the second module comprises registration or location features.
13. A device according to any of the above claims where the first module comprises registration or location features.
14. A device according to any of the above claims that further comprises a pressing element or elements arranged to exert localised forces through the protective membrane.
15. A device according to claim 14 where the pressing element or elements obstruct channels located in the first part, module or sub-assembly.
16. A device according to claim 14 where the pressing element or elements squeeze wicking materials located in the first part, module or sub-assembly.
17. A device according to claim 14 where the pressing element or elements initiate fluid movement through the creation of differential pressure.
18. A device according to claim 14 where the pressing element or elements cause electrical connections to be made.
19. A device according to any of the above claims where the protective enclosure membrane forms a partition between two chambers, one in each part, module or sub-assembly, in order to communicate pressure without the flow of any fluids.
20. A controller that receives the device according to claims 9 that causes said protective enclosure to be punctured or perforated.
21. A controller that receives a device according to claims 12 that engages with said registration or location features in the second part, module or sub-assembly.
22. A controller that receives a device according to any of claim 14 to 18 that causes said pressing element or elements to exert a force on the protective enclosure.
23. A controller that receives a device according to claim 10 that makes electrical connection to said discrete sensing element or fluid manipulation platform.
24. A manufacturing method for the assembly of a device accoiding to any of claims ito 19 comprising first and second module retention mechanisms.
25. A manufacturing method according to claim 23 where said first and second module retention mechanisms interleave or overlap to effect the handover of the retention of the device according to any of claims 1 to 19 from said first module retention mechanism to said second module retention mechanism.
26. A manufacturing method according to claim 23 or 24 where a first sheet of protective enclosure membrane material is attached to the first part, module or subassembly of said device and a second sheet of protective enclosure membrane material is attached to said first sheet.
27. A manufacturing method according to claim 25 where said second sheet of protective enclosure membrane material is attached to said first part, module or sub-assembly of said device in addition to said first sheet of protective membrane enclosure material.
28. A manufacturing method according to any of claims 23 to 26 that engages with registration or location features in the device according to claim 13.