Method for manufacturing a biological analysis card comprising a heating block

Abstract

The present invention is a method for manufacturing a biological analysis card (1), comprising: - at least two films (3) placed one on top of the other; - a receiving space (4) formed between the two films (3); - a step a) of preparing a base (2) formed from a duct (5) leading to the receiving space (4); - a step b) of placing a plate (9) into the receiving space (4), the plate (9) having: - a first face (10a) and a second face connected by an edge (11) having a profile; - a plurality of wells (12) opening onto at least the first face (10a) or the second face; - a plurality of channels (13) in fluid connection with the wells (12); - a welding step c) of welding the two films (3) within the receiving space (4) to the plate (9) and the films (3) to the edge (11) segments of the plate (9) by applying a heating block to the films (3) against the edge (11) on the side of the first face (10a).

Description

Technical Field

[0001] The present invention relates to the field of biological sample analysis, and more particularly to a method for manufacturing an analysis card using a heating block, an analysis card obtained by the method, and a heating block used therefor.

Background Art

[0002] In vitro biological analysis of biological samples such as polymerase chain reaction (PCR) tests is based on one or more reactions between a biological sample and one or more reagents. Biological samples can include tissues and cells derived from the human or animal body and their derivatives, organs, blood, its components or derived products. There are microfluidic systems and methods for performing these biological sample analyses. The reagents are placed in wells of a plate called an "array", and the biological sample to be analyzed is brought into contact with the reagents by flooding. The plate is assembled on a flexible base that includes a biological sample reservoir and ducts. The ducts in the base are connected to channels in the plate to form a fluid path. An analysis card such as a "FilmArray®" analysis card for performing a PCR test is formed. Assembling this card is a complex procedure that requires bonding at least four layers of film and adhesive layer to the plate before attaching the plate to the base. This involves the use of special machinery and a large amount of consumables (film and adhesive layer), which significantly increases the cost and increases the risk of defects in the case of leakage between the layers.

[0003] Furthermore, in the "FilmArray (registered trademark)" assay card configuration, only one biological sample reaches the plate via the duct. Therefore, "FilmArray (registered trademark)" cannot be used for the analysis of several biological solutions within the same assay card, for example, the same biological sample at different concentrations, and thus, for example, in the case of an endotoxin detection test. In an endotoxin detection test, the biological sample is diluted to different concentrations and each dilution is analyzed. Therefore, great care must be taken to ensure that different dilutions of the biological sample do not mix with each other before reaching the reaction wells of the plate so that the reaction results can be usable.

[0004] Finally, taking the endotoxin detection test as an example again, one specific aspect that must be pointed out is the plate thickness that is much larger than the plate thickness of "FilmArray (registered trademark)". The detection sensitivity required for endotoxin often requires a larger amount of reagents than the reagents required for a PCR test, which affects the thickness of the reaction wells and thus the thickness of the assay card, and thus this must be larger (the intensity of the fluorescent signal detected is directly proportional to the thickness of the fluorescent liquid present in the reaction well). Sealing the plate surface between the base films, as in the case of "FilmArray (registered trademark)", does not guarantee sealing at the edges of the plate and thus there is a risk of mixing of biological samples or other liquids, and this particular aspect causes problems with regard to sealing the plate at the base.

[0005] Therefore, in the context of, for example, an endotoxin detection test, there is currently no manufacturing method that enables obtaining an assay card with a thick plate between two base films, which is rapid, easy to manufacture, and inexpensive. Summary of the Invention

[0006] Accordingly, an object of the present invention is to manufacture an analysis card provided with a plate having a predetermined thickness capable of ensuring separation of fluid paths extending in the analysis card, and to analyze, for example, the same biological sample or different biological samples or several dilutions of different biological samples containing several dilutions more quickly, more easily, and more inexpensively in a more controlled manner without the risk of mixing.

[0007] According to a first aspect, a method for manufacturing a biological analysis card configured for biological analysis of a biological sample, comprising: - step a) of supplying a base composed of the following: - at least two superposed films, - an accommodation space formed between two films delimited by a first boundary and an opening, - a duct delimited by a contour and opening into the accommodation space through the first boundary, - step b) of inserting a plate into the accommodation space by inserting the plate through the opening, the plate comprising: - a first surface and a second surface connected by a rim having a profile, - a plurality of wells opening on at least the first surface or the second surface, - a plurality of channels fluidly connecting the wells and step b), - a welding step c) including welding the two films of the accommodation space to the plate and including, In welding step c), the film is welded to the rim segment of the plate by applying a heating block to the film against the rim on the side of the first surface, the heating block comprising a support and a heating element, a first part of the heating element protruding from the support at least during application of the heating block, and then the first part of the heating element having a profile complementary to at least one segment of the profile of the rim of the plate, such that during application of the heating block, the film assumes the shape of the rim when the film is pressed by the first part of the heating element. A method is proposed.

[0008] According to advantageous but non-limiting features which can be carried out alone or in any combination, - the profile of the rim segment of the plate is a straight line defining a convex angle between the segment of the rim of the plate and a second surface which is at most 90° thereto, or a curve whose tangent defines a convex angle between the segment of the rim of the plate and a second surface which is at most 90° thereto; - the rim segment to which the film is welded is a separating surface located on the rim between channels; - the first part of the heating element of the heating block includes a non-heating zone whose width is greater than the width of the channel such that when the first part of the heating element is applied to the film against the rim segment, the non-heating zone faces the channel and the fragile valve; - each duct is equipped with at least one fragile valve positioned at one end of the duct; - step c) includes welding the film by means of the heating block up to the duct-to-duct boundary separating the duct from the segment of the rim of the plate; - step b) includes positioning the plate in the receiving space until the mouth of the channel faces the mouth of the duct; - step c) includes closing the opening of the receiving space by welding two films to each other; - The plate comprises a vacuum port connected to a channel, the channel being connected to a well, and the method includes a vacuum step after welding step c), the vacuum step including perforating a film covering the vacuum port and, once the vacuum port is created, closing the channel connecting the vacuum port to the well; - The thickness of the plate is greater than 0.5 mm; - Welding two films of the receiving space to the plate includes welding a first film to a first face; - The heating block comprises a second part of a heating element having a shape complementary to the shape of the first face of the plate, and welding two films of the receiving space to the plate includes welding a first film to the first face, the first film being pressed against the first face.

[0009] According to a second aspect, an analysis card manufactured by the above method is proposed.

[0010] According to a third aspect, a heating block configured to carry out the above method is proposed, the heating block comprising a support and a heating element, a first part of the heating element being configured to project at least during application of the heating block, the first part of the heating element having a profile complementary to at least a part of the profile of the rim of the plate.

[0011] This heating block is advantageously complemented by the following various features, which may be implemented alone or in various possible combinations thereof: - The first part of the heating element includes a non-heating region whose width is greater than the width of the channel; - The non-heating zone of the first part of the heating element is a recess; - The first part of the heating element includes a flexible nickel-chromium (NiCr) element.

[0012] Other features, objects, and advantages of the present invention, which are purely illustrative and non-limiting, will become apparent from the following description, which must be read with reference to the accompanying drawings:

Brief Description of the Drawings

[0013]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

DETAILED DESCRIPTION OF THE INVENTION

[0014] The present invention relates to a method for manufacturing the analysis card 1 shown in FIG. 4. FIG. 1 provides details of the steps of the method, which includes, first, step a) of supplying the base 2. Referring to FIG. 2, the base 2 is formed from two thermoplastic resin films 3 that are overlapped. The material constituting the film 3 may be, for example, a composite based on polypropylene or polyethylene. The film 3 has a thin and flexible surface. "Thin" means that the thickness of the film 3 is less than 200 μm. Preferably, the film 3 has a rectangular shape. More preferably, the two films 3a, 3b have the same shape when overlapped. Therefore, since one is on top of the other, the two films 3a, 3b in FIG. 2 cannot be distinguished. Film 3b is referred to in FIG. 5. The two films 3 are thermoplastic resins, which means that when heated to a certain temperature threshold, the film 3 softens and can undergo mechanical deformation that is fixed when the film 3 cools. The temperature threshold at which the film 3 can soften and deform depends on the material constituting the film 3. The material constituting the film 3 is preferably a polymer such as polypropylene, polyethylene, or a composite film based on polypropylene or polyethylene. Typically, the temperature threshold required to soften the film 3 exceeds 60°C.

[0015] An accommodation space 4 is formed between the two films 3. More specifically, the accommodation space 4 is included between two advantageous film 3 parts. "Free" means that the films 3 are not welded to each other. In the plane of the base 2, the accommodation space 4 is delimited by a first boundary 7 and an opening 8. The first boundary 7 is an area where the two films 3 are welded together. In the example shown in FIG. 2, the first boundary 7 forms a straight line parallel to the width of the film 3, along which the two films 3 are welded to each other. This welding may be carried out, for example, using a laser that melts the film 3 along this path so that the two films 3 are welded to each other along an exact path. Other boundaries where the two films 3 are welded to each other can delimit the accommodation space 4.

[0016] The opening 8 corresponds to an area where the films 3 are not welded to each other and remain free up to their edges. The opening 8 enables access to the accommodation space 4 from the outside. Preferably, the accommodation space 4 has the same shape as the plate to be accommodated, typically rectangular.

[0017] The opening 8 is added to the first boundary 7 and may include one or more of three different edge portions of the film 3 located on the right side of FIG. 2. Thus, in this case, the opening 8 includes an edge of the film 3 parallel to the first boundary 7 and two edges of the film 3 perpendicular to the first boundary 7. These edge portions of the film 3 are free and not welded to the edge portions of another film 3. According to the preferred embodiment shown in FIG. 2, the first boundary 7 and the second boundary 230 of the base 2 form the pocket 23 such that the opening 8 is located on only one side of the film 3.

[0018] The base 2 is formed between two films 3 and further includes a duct 5 delimited by a contour 6. The duct 5 is like a tunnel formed between two films 3 and enables the circulation of a fluid such as a biological liquid sample. The contour 6 of the duct 5, like the first boundary 7, forms a curve where the two films 3 are welded to each other. Each duct 5 is delimited by two contours 6. The contours 6 of different ducts 5 are connected by an inter-duct boundary 7a where the two films 3 are welded to each other. The inter-duct boundary 7a forms part of the first boundary 7. The base 2 may also include various elements such as a reservoir formed between the films of the base 2 and connected to the duct 5. Each duct 5 may be provided with one or more frangible valves that close the duct 5. For example, these frangible valves may be defined by an area (measuring a few millimeters or centimeters) where the film 3 is weakly welded and configured to break as a function of the pressure applied to the frangible valve. The frangible valves are preferably located near the receiving space 4, in other words, at the end of the duct 5 that opens into the receiving space 4. For example, a reservoir that may take the form of a blister may contain a biological sample. The reservoir may be pressurized until the pressure applied to a frangible valve present in the duct 5 connected to the reservoir reaches a certain pressure threshold required for the frangible valve to rupture. When the frangible valve ruptures, the biological sample contained in the reservoir flows through the duct 5 to the receiving space 4.

[0019] The manufacturing method then includes step b) of placing a plate 9 inside the base 2 so as to finally have the base 2 with the plate 9 as shown in FIG. 3. The plate 9 generally has a flat surface and a certain thickness, but is defined as an element that is very small compared to the dimensions of its flat surface. For example, the thickness is at least one tenth smaller than the width and length of the surface. Preferably, the plate 9 has a thickness of 0.5 mm or more. In the example shown, the plate 9 has a rectangular surface, but other shapes may be selected.

[0020] Preferably, the plate 9 is formed from a thermoplastic resin material, at least on its surface. For example, the plate 9 may be made of materials such as PP, PE, PMMA, PC, PS, POM, ABS, COP, etc. As schematically shown in FIG. 5, the plate 9 is composed of two surfaces 10a, 10b that face each other and are separated by a small thickness, that is, the thickness is at least one-tenth smaller than the width and length of the surfaces 10a, 10b. The maximum and minimum dimensions of the surfaces 10a, 10b of the plate 9, in this case the length and width of the surfaces 10a, 10b, are preferably greater than 2 cm, also preferably less than 10 cm, more preferably less than 4 cm. The thickness of the plate 9 is preferably less than 5 mm, more preferably less than 3 mm, and the thickness of the plate 9 preferably exceeds 0.5 mm. The surfaces 10a, 10b are connected by side surfaces including a rim 11. In other words, the rim 11 corresponds to another surface of the plate 9 that has a small width, that is, at least one-tenth smaller than the width and length of the surfaces 10a, 10b. Therefore, the length of the rim 11 is equal to the length or width of the surfaces 10a, 10b of the plate 9. Preferably, the profile of the rim 11 is inclined with respect to the surfaces 10a, 10b and thus is not perpendicular to the surfaces 10a, 10b. Preferably, referring to FIG. 6, the profile of the rim 11 is a straight line that defines a convex angle θ of less than 90° between the rim 11 of the plate 9 and the second surface 10b. In other words, the smaller of the two angles defined by the profile of the rim 11 between the rim 11 and the second surface 10b, that is, the angle less than 180°, is less than 90°. In other words, the rim 11 preferably forms an inclined surface. According to another embodiment shown in FIG. 7, the profile of the rim 11 is curved, and its tangent defines a convex angle θ of less than 90° between the rim 11 of the plate 9 and the second surface 10b. The tangent is shown, for example, as a dotted line in FIG. 7. In other words, the smaller of the two angles defined by the profile of the rim 11 between the rim 11 and the second surface 10b, that is, the angle less than 180°, is less than 90°. According to this embodiment, the profile of the rim 11 is preferably a convex curve.

[0021] Plate 9 comprises a plurality of wells 12 that open onto at least one of the faces 10a, 10b of the plate 9. The wells 12 can also pass through the plate 9 from one face 10a to the other face 10b. The wells 12 can contain a reagent 27. For example, in the case of a test for detecting the presence of endotoxin, the wells 12 contain three different reagents 27, an activator for activating a detector, an enzyme and a fluorogenic substrate that is in an inactive state in the absence of activation in the absence of endotoxin, and a control reagent 27 adapted to control the functionality of the detection reagent 27. The wells 12 are connected by channels 13 configured to supply a liquid biological sample or another liquid such as a reference fluid used, for example, in a control well 12 to the wells 12. The channels 13 are delimited by two edges 13a, 13b. It should be noted that the edges 13a, 13b constitute a contact surface between the interior of the channel 13 and the face 10a of the plate 9. The edges 13a, 13b extend opposite to each other in the direction in which the channel 13 extends. In Figure 3, the edges 13a, 13b cannot be distinguished from the channel 13 and it is specified that the channel 13 is very thin and is shown using a line. The edges 13a, 13b are referred to in Figure 8. The channels 13 are present on both the face 10a of the plate 9 and on the rim 11 of the plate 9. In other words, it can be stated that the channels 13 extend from the rim 11 to the face 10a or that the channels 13 extend from the face 10a to the rim 11.

[0022] The plate 9 is placed into the receiving space 4 of the base 2. This space (or functional clearance) is necessary to easily insert the plate 9 into the base 2. More specifically, the plate 9 is inserted into the receiving space 4 via the opening 8. The plate 9 is arranged such that the rim 11 of the plate 9 faces the first boundary 7 of the base 2. Preferably, referring to FIG. 1, the step b) of placing the plate 9 into the receiving space 4 includes the step b1) where the mouth of the channel 13 of the plate 9 is positioned on the opposite side of the mouth of the duct 5 of the base 2. Thus, each channel 13 opens on the opposite side of the mouth of the duct 5 such that each channel 13 forms a fluid path with the duct 5. As a result, the plate 9 is considered to be correctly positioned within the receiving space 4 when the mouth of the channel 13 is at the center of the mouth of the duct 5 and thus a fluid path is formed. Thus, the fluid connection between the channel 13 and the duct 5 is preferably ensured at the rim 11 of the plate 9, which enables the analysis card 1 to be obtained without being too bulky and with easier packaging. The plate 9 can be automatically inserted into the receiving space 4 using a clamp that grips the plate 9 and inserts the plate 9 into the receiving space, and the gripping area of the clamp enters the receiving space 4. As described above, according to a preferred embodiment, the first boundary 7 and the second boundary 230 form a pocket 23 within the base 2. A "pocket" means a receiving space whose boundaries 7, 230 are longer than the opening, more specifically, a container that opens only on one side of the base 2, is formed by the film 3, and is delimited by the first boundary 7 and the second boundary 230. Thus, the placement of the plate 9 includes the insertion of the plate 9 into the pocket 23. Preferably, the fitting of the pocket 23 is such that the plate 9 cannot be inverted or reoriented when the base 2 equipped with the plate 9 is moved. In other words, except for being along the opening 8, the first boundary 7 and the second boundary 230 surround the plate 9, and when the plate 9 is placed within the receiving space 4, the receiving space 4 is not mostly left empty.

[0023] Next, a welding step c) is performed, which includes welding two films 3 of the accommodation space 4 to the plate 9. Preferably, step c) includes welding the film 3 to the surface 10 of the plate 9. Accordingly, the two films 3 are welded to the surfaces 10a, 10b of the plate 9 in the region of the accommodation space 4. In other words, the first film 3a is welded to the surface 10a, and the second film 3b is welded to the surface 10b. This welding may be carried out using a heating object that melts the film 3 on the plate 9. Accordingly, the film 3 is welded to the surfaces 10a, 10b of the plate 9. For example, the heating object may have a surface with a dimension at least equal to one of the dimensions of the surfaces 10a, 10b. When pressed against the surface 10a or 10b, the surface of the heating object melts the films 3a, 3b disposed on the surface 10a or 10b, and the films 3a, 3b are welded to the surface 10a or 10b. It will be understood that when this welding is completed, the plate 9 is fixed to the base 2. The plate 9 may no longer need to move within the accommodation space 4.

[0024] Advantageously, the welding of the film 3a to the surface 10a and the welding of the film 3b to the surface 10b are carried out simultaneously. This can improve the method in various respects. First, the welding process is carried out more quickly. Second, the plate 9 placed on the base 2 is inserted into the welding machine only once. If step c) includes two steps of welding the film 3 to the surface 10 of the plate 9 (one for the first film 3a and the other for the second film 3b), when one film 3 is welded to one surface 10, the plate 9 and the base 2 must be turned over in order to weld the other film 3 to the other surface 10. By inserting the plate 9 and the base 2 into the welding machine only once, it is possible to speed up the method and limit potential positioning errors. Third, since the plate 9 and the base 2 are positioned in exactly the same way for the welding of the film 3a to the surface 10a and the welding of the film 3b to the surface 10b, the welding becomes more uniform (because these two weldings are carried out simultaneously). To weld the two films 3 on the surface 10 of the plate 9 simultaneously, two heating objects are applied simultaneously. The first heating object is applied to the first film 3a and the first surface 10a, and the second heating object is applied to the second film 3b and the second surface 10b. According to a particular embodiment, the heating objects are made of nichrome and are not heated before being applied to the films 3a, 3b and the surfaces 10a, 10b. In use, these nichrome heating objects are first pressed against the films 3a, 3b and the surfaces 10a, 10b, and then current pulses are sent to the nichrome heating objects to induce rapid heating of the nichrome heating objects.

[0025] According to another embodiment, the welding of the film 3a to the surface 10a and the welding of the film 3b to the surface 10b are carried out continuously.

[0026] Welding step c) includes welding the first film 3a to the rim 11 segment of the plate 9 by applying the heating block 14 to the first film 3a against the rim 11 on the side of the first surface 10a. The welding of the first film 3a to the rim 11 segment of the plate is preferably performed simultaneously with or after the welding of the first film 3a on the surface 10 of the plate 9.

[0027] When the film 3 is welded only to the surface 10 of the plate 9, the rim 11 of the plate 9 remains free, that is, the film 3 is not welded to the rim 11. As a result, the biological samples circulating in the channel 13 can be mixed with each other at the rim 11, and each biological sample can circulate out of the channel 13 and into another channel 13. As described above, if it is desired to obtain valid and usable reaction results, such mixing of different biological samples before reaching the reaction well 12 cannot be suppressed. For example, in the case of an endotoxin detection test, biological samples must be analyzed at different concentrations. These fluids must not be mixed before reaching their respective wells 12, otherwise the results of the reaction cannot be used. Therefore, it is necessary to weld the film 3 to the rim 11 segment so as to strengthen the attachment of the plate 9 to the base 2 and isolate the channel 13.

[0028] Preferably, the rim 11 segment to which the film 3a is welded is a separation surface 18 located on the rim 11 between the channels 13, as shown in FIG. 8. The separation surface 18 is shown in FIG. 8 as a surface filled with dashes. Thus, in the rim 11, the channels 13 are separated by the separation surface 18. Thus, here it will be understood that the separation surface 18 is a rim 11 segment and thus part of the plate 9. The separation surface 18 has an arbitrary shape and is delimited by at least one second side surface 180. Preferably, the separation surface 18 has a rectangular shape and is thus delimited by four side surfaces 180a to 180d. The separation surface 18 may extend longitudinally on the rim 11 in the direction x shown in FIG. 8 from the edges 13a, 13b of the first channel 13 to the edges 13b, 13a of the second channel 13, and the second channel 13 is continuous with the first channel 13. Preferably, the separation surface 18 does not contact the channels 13 and is thus located between a first boundary fixed at a first distance from the edges 13a, 13b of the first channel 13 and a second boundary fixed at a second distance from the edges 13b, 13a of the second channel 13. In other words, there is a virtual margin around the channels 13 where the separation surface 18 does not break. As shown in FIG. 8, the side surfaces 180a and 180c do not contact the edges 130b, 131a of the channels 130, 131. Preferably, the separation surface 18 is rectangular and thus more preferably, as shown in FIG. 8, the side surfaces 180a and 180c are repeated to be parallel to the edges 130b, 131a of the channels 130, 131. More specifically, as schematically shown, the side surface 180a is located in the virtual margin of the edge 130b, and the side surface 180c is located in the virtual margin of the edge 131a. With respect to the direction y shown in FIG. 8, the separation surface 18 does not extend beyond the rim 11. Thus, the separation surface 18 does not extend to the surface 10a of the plate 9 or to the surface 10b of the plate 9. In other words, the side surfaces 180b and 180d are located between the contact surface with the surface 10a of the rim 11 and the contact surface with the surface 10b of the rim 11. Preferably, the side surfaces 180b and 180d are parallel to the contact surface with the surface 10a of the rim 11 and the contact surface with the surface 10b of the rim 11.More preferably, the side surface 180b coincides with the contact surface with the surface 10a of the rim 11, and the side surface 180d coincides with the contact surface with the surface 10b of the rim 11.

[0029] Welding of the film 3 to the rim 11 segment is carried out using the heating block 14 shown in FIG. 9. The heating block 14 comprises a support 15 and a heating element 16. The heating element 16 is set to a heating temperature suitable for at least partially melting the film 3, for example at least 60°C. The support 15 and the heating element 16 can be two independent components connected to each other. For example, the support 15 and the heating element 16 may be adhesively bonded or welded. The heating element 16 is made of a material that conducts heat and is typically made of metal. The heating element 16 includes a first portion 160 that projects along the rim 11 substantially from one surface 10a to the other surface 10b at least while the heating block 14 is applied to the rim 11. In the illustrated example, the first portion 160 projects from the support 15 so as to form a ledge from the support 15.

[0030] The first part of the heating element 160 has a profile that is complementary to at least a part of the profile of the rim 11 of the plate 9. Thus, the first part of the heating element 160 conforms to the shape of the rim 11 segment. If the rim 11 has an inclined shape, the first part of the heating element 160 has a shape complementary to this inclined part so that the rim 11 segment can fit against the first part of the heating element 160. Thus, when the heating block 14 is applied to the film 3a disposed on the rim 11 of the plate 9, the film 3a melts by the heat from the heating element 16 and the first part of the heating element 160 is welded to the rim 11 segment having the complementary profile. Preferably, the profile of the first part of the heating element 160 is complementary to the separation surface 18 of the rim 11. In other words, the heating block 14 comprises a non-heating zone 26 whose width is greater than the width of the channel 13. The non-heating zone 26 may be a recess 26 in the first part of the heating element 16. These recesses 26 may take the form of grooves, for example. The non-heating zone 26 may also be a part made of an insulating material that does not conduct heat in the first part of the heating element 160. When the heating element 16 is applied to the rim 11 of the plate 9, the non-heating zone 26 is positioned in the channel 13 so that a part of the heating element 16 is not directly applied to the channel 13. As a result, the heating element 16 does not weld the film 3a to the channel 13 but welds the film 3a only to the separation surface 18 and thus between the channels 13 of the rim 11. Thereby, the channels 13 can be specifically isolated from each other and the circulation of biological samples between the various channels 13 in the rim 11 can be prevented.

[0031] As in the illustrated embodiment, the heating element may be rigid, in which case the first part of the heating element 160 permanently protrudes and always has a contour complementary to the profile of the rim 11. It is also possible that the heating element 16 is flexible, and thus the protrusion of the first part of the heating element 160 is caused by the application of the heating element 16 to the rim 11. For example, the first part of the heating element 160 may comprise a strip or bundle of metal wires made, for example, from nichrome (NiCr). The flexibility of the heating element 16 allows for self-adjustment to compensate for any small geometric differences that occur, for example, during the manufacturing process of the plate 9 by injection molding.

[0032] According to a preferred embodiment, the welding of the film 3 to the rim 11 segments of the plate 9 extends at least to the inter-duct boundary 7a that separates the ducts 5. Thus, it will be understood that the welding of the film 3 extends continuously from the rim 11 segments at least to the inter-duct boundary 7a. In other words, step c) includes welding the film 3a to the transition sub-zone 20 of the transition zone 19 of the film 3b that delimits the transition space 25. Referring to FIG. 3, when the plate 9 is fixed within the base 2, a transition space 25 is formed between the rim 11 and the first boundary 7. The transition space 25 is defined between a part of the film 3a and a part of the film 3b, and the part of the film 3b is the transition zone 19. Thus, the transition zone 19 is the part of the film 3b that is not welded to the film 3a or the plate 9 between the rim 11 and the first boundary 7. In FIG. 3, the transition zone 19 is a stripe-like surface. The transition zone 19 is located at an extension of the separation surface 18 and includes a transition sub-zone 20 that extends to the inter-duct boundary 7a. FIG. 8 shows an enlarged view of FIG. 3. The transition sub-zone 20 is shown in FIG. 8 and is a dotted-line surface within the transition zone 19 that itself is stripe-like in FIG. 8. The transition sub-zone 20 is not located on the opposite side of the mouth of the channel 13 or the duct 5 within the transition zone 19. On the contrary, the transition sub-zone 20 is located between the fluid paths. Note that the fluid paths are formed by the channels 13 and ducts 5 that open to opposite sides of each other. Thus, each fluid path includes the channel 13, the duct 5, and a part of the transition zone 19, and the biological sample circulating within the duct 5 can reach the channel 13 through this part of the transition zone 19.

[0033] It should be noted that the transition space 25 is part of the accommodation space 4 located between the rim 11 of the plate 9 and the first boundary 7 of the base 2. This transition space 25 poses problems for the analysis of different biological samples. It is not desirable to leave the transition space 25 in a free state because this allows the mixing of different biological samples circulating from the duct 5 of the base 2 to the channel 13 of the plate 9. Before the film 3 is welded to the surfaces 10a, 10b, that is, before the contact surface between the rim 11 of the plate 9 and the surface 10b is joined to the first boundary 7, it should be understood that the transition zone 19 and thus the volume of the transition space 25 still exist even if the plate 9 is arranged to minimize the surface area of the transition zone 19, and hence the volume of the transition space 25. This is firstly because the film 3 is flexible and the transition zone 19 and the transition space 25 are narrow but remain when the film 3 is welded only to the surface of the plate 9. It is also explained by the fact that when the film 3 is welded to the surfaces 10a, 10b, there is no guarantee that the plate 9 will not move slightly within the accommodation space 4. Therefore, it is preferable to weld the transition subzone 20 of the film 3b to the film 3a. Thus, the film 3a is welded to the transition subzone 20 and thus the films 3a and 3b are welded to each other between the fluid paths, allowing the circulation of biological samples between the duct 5 and the channel 13 of each fluid path 26 while preventing the circulation of biological samples between the duct 5 and the channel 13 of different fluid paths 26. Thus, by welding the films 3a and 3b together up to the duct boundary 7a, it is possible to enhance the isolation of the various fluid paths so that the biological samples circulating from the duct 5 to the channel 13 do not mix in the transition zone 19. In this preferred embodiment, the first portion of the heating element 160 has a profile with a shape complementary to both the rim 11 segment of the plate 9 and the portion of the base 2 within the transition subzone 20.

[0034] More preferably, the welded portion of the film 3 extends beyond the duct-to-duct boundary 7a. In other words, the welding step c) includes welding the film 3 to the plate 9 from the rim 11 segment of the plate 9 beyond the duct-to-duct boundary 7a that separates the ducts 5. Thus, this means that in the zone 21 of the film 3b schematically shown using the grid pattern, the films 3a and 3b are welded to each other between the ducts 5, as shown in FIG. 8. By extending the welding beyond the duct-to-duct boundary 7a, it is possible to ensure that the films 3a and 3b are completely welded to each other from the rim 11 segment to the duct-to-duct boundary 7a. In this preferred embodiment, the first portion of the heating element 160 has a profile with a complementary shape beyond the duct-to-duct boundary 7a with respect to both the rim 11 portion of the plate 9 and the portion of the base 2 within the transition subzone 20. Preferably, the duct 5 is not heated to avoid rupture of the frangible valve.

[0035] Advantageously, the step of welding the film 3a to the face 10a of the plate 9 and the step of welding the film 3 to the rim 11 segment of the plate 9 are carried out simultaneously using the same heating block. In this case, the heating element 16 of the heating block 14 includes a second portion having a shape complementary to the shape of the face 10 of the plate 9. Thus, when the heating element 16 of the heating block 14 is applied to the film 3a covering the face 10a of the plate 9, the second portion of the heating element conforms to the shape of the face 10a of the plate 9, welding the film 3a to the face 10a, and the first portion of the heating element 160 welds the film 3a to the rim 11 segment of the plate 9. Welding the film 3a to the face 10a simultaneously with welding the film 3a to the rim 11 segment has several advantages. First, since the steps are not separate but are carried out simultaneously, the method is speeded up. Also, the number of rejections is reduced because the problems of the operation of the machine and the positioning of the plate 9 and the base 2 within the machine are less likely to occur in one step rather than in two steps. Further, since no dedicated tools are required for welding the film 3a to the face 10a and for welding the film 3a to the rim 11 segment, the number of tools required is reduced, only the heating block 14 is present, and the cost is reduced. Finally, by heating the plate 9 only once, the risk of degradation of the heat-sensitive reagent 27 already present in the well 12 is reduced.

[0036] More preferably, the welding of the first film 3a to the first face 10a, the welding of the second film 3b to the second face 10b, and the welding of the first film 3a to the rim 11 segment are carried out simultaneously. Thus, it will be understood that the welding of the first film 3a to the first face 10a and the welding of the first film 3a to the rim 11 segment are carried out using a single heating block 14. The welding of the second film 3b to the second face 10b may be carried out using another object to be heated. The heating block 14 is applied to the first film 10a at the same time as the object to be heated is applied to the second film 10b. Thus, according to this embodiment, the method is even faster.

[0037] According to another embodiment, particularly when the plate 9 is thin (i.e., less than 0.5 mm), the profile of the rim 11 of the plate 9 may be perpendicular to the surfaces 10a, 10b and not inclined with respect to the surfaces 10a, 10b. In this case, the films 3a, 3b are welded to the rim 11 by rivet feeding.

[0038] Preferably, the method includes a step of closing the receiving space 4 so that the receiving space 4 is no longer open to the outside through the opening 8. The closing step is carried out by welding the two films 3 together. The welding may be carried out, for example, using a laser or by applying a heating object to the film 3. FIG. 4 showing the analysis card 1 obtained at the end of the method shows a welding line 40 closing the receiving space 4 of the pocket 23. By first closing the receiving space 4, it is ensured that the plate 9 does not come out of the receiving space 4 and thus remains within the receiving space 4. This also serves to isolate the plate 9 and prevent the plate 9 from being damaged. This also protects the plate 9 from the outside, thereby preventing any foreign particles or foreign bodies that would impair the effectiveness of the reaction in the well 12 upon arrival of the biological sample from entering the well 12 of the plate 9. Preferably, the welding of the two films 3 closing the receiving space 4 closely surrounds the plate 9 so that the plate 9 is firmly held and immobilized inside the base 2. Thus, preferably, in FIG. 3, the weld closing the opening 8 extends along the side surface of the plate 9, and more preferably, the distance separating the weld closing the opening 8 and the plate 9 is less than 5 mm, more preferably less than 2 mm.

[0039] A method for manufacturing the analysis card 1 preferably includes the step of placing the plate 9 under reduced pressure. The step of placing the plate 9 under reduced pressure aims to create a vacuum in the wells 12 and channels 13 of the plate 9. When a vacuum is generated in the plate 9, any biological sample introduced into the duct 5 of the base 2 is sucked into the channel 13 and then into the wells 12 of the plate 9. Thus, a reaction occurs between the biological sample and the reagent 28 in the wells 12. Further, the vacuum step may be performed before the step of welding the film 3 to the surface 10 of the plate 9. Thus, when the film 3 is welded to the surface 10 of the plate 9, there are no air bubbles between the film 3 and the surface 10.

[0040] To perform this vacuum step, referring to FIG. 4, the supplied plate 9 preferably has a vacuum port 24. The vacuum port 24 is connected to the channel 13 of the plate 9. Preferably, the vacuum port 24 opens on at least one surface 10a of the plate 9. A vacuum member such as a suction cup adheres to the vacuum port, and a vacuum is generated in the channels 13 and wells 12 of the plate 9. The operation can last from 15 seconds to 2 minutes, usually 30 seconds. Next, a heating member is applied to the channel 13 connected to the vacuum port 24 to melt the plate 9 in the channel 13 and block the channel 13. Thus, the vacuum is trapped in the wells 12 and channels 13 of the plate 9.

[0041] When a vacuum is generated, according to an embodiment where the film 3 is already welded to the surface 10, the vacuum step includes perforating the film 3 covering the vacuum port 24. Specifically, it is essential to access the vacuum port 24 to suck out the air and create a vacuum in the plate 9. Further, it is preferable that the duct 5 is not open to the outside so that the vacuum is effective. Thus, for example, the duct 5 may include a fragile valve that closes the duct 5 at least during the vacuum step.

[0042] The present invention is not limited to the embodiments described and illustrated in the accompanying drawings. Without departing from the scope of protection of the present invention, changes can be made, particularly from the perspective of the nature of various technical features or the substitution of technical equivalents.

Claims

1. A method for manufacturing a biological analysis card (1) configured for the biological analysis of a biological sample, - Step a: Supplying a base (2) consisting of the following: - At least two superimposed films (3), - A containment space (4) formed between two films (3) separated by a first boundary (7) and an opening (8), - A duct (5) separated by a contour (6) and opening into the containment space (4) through the first boundary (7), - Step b) of inserting the plate (9) into the storage space (4) by inserting the plate (9) through the opening (8), - A first surface (10a) and a second surface (10b) connected by a rim (11) having a certain profile, - A plurality of wells (12) opening on at least the first surface (10a) or the second surface (10b), - Multiple channels (13) that fluidly connect the wells (12) including step b), - A welding step c) including welding the two films (3) of the containment space (4) to the plate (9) Includes, Welding step c) includes welding the film (3) to a segment of the rim (11) of the plate (9) by applying a heating block (14) to the film (3) against the rim (11) on the side of the first face (10a), wherein the heating block (14) comprises a support (15) and a heating element (16), wherein a first portion of the heating element (160) protrudes from the support (15) at least during the application of the heating block (14), and the first portion of the heating element (160) has a profile complementary to at least one segment of the profile of the rim (11) of the plate (9), and as a result, the film (3) takes the shape of the rim (11) when pressed by the first portion of the heating element (160) during the application of the heating block (14), method.

2. A method for manufacturing an analysis card (1) according to claim 1, wherein the profile of a segment of the rim (11) of the plate (9) is a straight line defining a convex angle (θ) of 90° or less between the segment of the rim (11) of the plate (9) and the second surface (10b), or the tangent thereto is a curve defining a convex angle (θ) of 90° or less between the segment of the rim (11) of the plate (9) and the second surface (10b).

3. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein the rim (11) segment to which the film (3) is welded is a separation surface (18) located on the rim (11) between channels (13).

4. A method for manufacturing an analysis card (1) according to claim 3, wherein the first portion of the heating element (160) of the heating block (14) includes a non-heated zone (26) whose width is greater than the width of the channel (13) such that the non-heated zone (26) faces the channel (13) and the fragile valve when the first portion of the heating element (160) is applied to the film (3) with respect to the rim (11) segment.

5. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein step c) includes welding a film (3) to an inter-duct boundary (7a) that separates the duct (5) from a segment of the rim (11) of the plate (9) using a heating block (14).

6. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein step b) includes positioning the plate (9) within the containment space (4) until the opening of the channel (13) faces the opening of the duct (5).

7. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein step c) includes the step of closing the opening (8) of the containment space (4) by welding two films (3) together.

8. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein a plate (9) comprises a vacuum port (24) connected to a channel (13), the channel (13) being connected to a well (12), and the method includes a vacuum step after a welding step c), the vacuum step comprising perforating a film (3) covering the vacuum port (24) and, once a vacuum is created, closing the channel (13) connecting the vacuum port (24) to the well (12).

9. A method for manufacturing the analysis card (1) according to claim 1 or 2, wherein the thickness of the plate (9) is greater than 0.5 mm.

10. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein welding two films (3) of a containment space (4) to a plate (9) includes welding a first film (3a) to a first surface (10a).

11. A method for manufacturing an analysis card (1) according to claim 1 or 2, wherein the heating block includes a second portion of a heating element whose shape is complementary to the shape of the first surface (10a) of the plate (9), and welding the two films (3) of the containment space (4) to the plate (9) includes welding the first film (3a) to the first surface (10a), so that the first film (3a) is pressed against the first surface (10a).

12. An analysis card (1) manufactured by the method described in claim 1 or 2.

13. A heating block (14) configured to perform the method of claim 1 or 2, wherein the heating block (14) comprises a support (15) and a heating element (16), wherein a first portion of the heating element (160) is configured to protrude at least during application of the heating block (14), and the first portion of the heating element (160) has a profile complementary to at least a portion of the profile of the rim (11) of the plate (9).

14. The heating block (14) according to claim 13, wherein the first portion of the heating element (160) comprises a non-heating zone (26) whose width is greater than the width of the channel (13).

15. The heating block (14) according to claim 14, wherein the non-heating zone (26) of the first part of the heating element (160) is a recess (26).

16. The heating block (14) according to claim 13, wherein the first portion of the heating element (160) includes a flexible nichrome (NiCr) element.

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