Device for holding tubes used for analysing samples by means of nucleic acid amplification techniques and method for using such a device

Abstract

The invention relates to a device comprising - a tray (7) for holding tubes receiving samples to be analysed, said tray (7) having holes (8) suitable for each receiving a tube bearing on the upper surface and with the bottom thereof protruding from the tray; - a cover (10) provided with flues (11) arranged in a distribution similar to the distribution of the holes of the tray, said cover (10) being designed to occupy at least one overlapping position above the tray (7), such that the cover can be joined in a removable manner to the tray in order to form a unitary assembly for holding the tubes and to position the flues (11) over the tubes to act as guides for pipette tips entering the tubes.

Description

Technical Field

[0001] The present invention relates to the technical field of devices used for handling tubes receiving biological samples to be analyzed.

[0002] The device according to the invention generally relates to the processing of samples for analysis, in the field of microbiology. It finds particularly advantageous applications in assisting in the handling of tubes receiving biological samples to be analyzed by nucleic acid amplification techniques such as the Polymerase Chain Reaction (or "PCR") and similar methods. Prior art

[0003] Nucleic acid amplification techniques are well known in the prior art. These techniques, and in particular PCR, implement different steps to prepare and analyze biological samples, including steps of rehydration of reagents, mixing of reagents with the sample, centrifugation, amplification and detection. This technique uses tubes that are pre-filled with reagents and each closed with a sealing cap. These tubes are intended to receive the biological samples to be analyzed. These tubes are manipulated by suitable devices in order to carry out the different steps of this technique.

[0004] In the prior art, there are numerous devices for ensuring the implementation of the PCR technique while allowing simple and safe handling of the tubes when carrying out the different stages of this technique.

[0005] For example, document CN 107868752 describes a device comprising a support plate for tubes of samples to be analyzed, intended to be supported by a base provided with a positioning base in an analysis machine. This device also comprises a cover provided with tabs intended to be magnetically fixed to the base, so that the tube support plate is immobilized between the base and the cover. Furthermore, this cover is provided with passage holes for a pipette tip, arranged in correspondence with the tubes. Such a device makes it possible to simultaneously handle a set of tubes for loading and positioning in the analysis machine.

[0006] However, such a device does not guarantee the immobilization of the tubes on the base. Indeed, the system of assembling the cover to the base by a magnetic force does not allow for the development of a sufficient assembly force when the tubes supported by such a device undergo, in particular, the centrifugation operation. Furthermore, the presence of the cover above the tubes makes the operation of depositing inside the tubes even more complex, which is a delicate operation to carry out since the pipette must ensure the deposit inside the tubes of a drop of small volume, typically 5 to 10 microliters.

[0007] US Patent 7,329,393 describes a device for supporting tubes of samples to be analyzed, adapted to undergo a centrifugation operation. Such a device comprises a cover intended to be fixed to a base by a screw assembly system and between which are interposed a tube receiving tray and a support block. Although such a device allows effective holding of the tubes while they undergo the centrifugation operation, the assembly and disassembly operations prove relatively time-consuming to carry out. It follows that the use of such a device is limited in practice to this centrifugation operation. Furthermore, such a device does not provide a solution to the problem of depositing samples inside the tubes.

[0008] Patent application US 2015 / 0132841 describes an assembly comprising a support for containers receiving samples to be analyzed and a lid provided with openings arranged in a distribution similar to the containers. The lid is temporarily assembled to the support using fixing elements such as elastically deformable arms rising from the support and cooperating with projections carried by the lid. Such an assembly makes it possible to hold the containers in position. However, such an assembly does not provide a solution to the problem of depositing samples inside the containers. Statement of the invention

[0009] An object of the invention is to remedy the drawbacks of the prior art by proposing a support device for tubes intended to receive reagents necessary for an amplification reaction and biological samples to be analyzed, such a device being capable of being implemented in a simple and efficient manner for different preliminary operations or during an amplification reaction.

[0010] An object of the invention is to propose a support device for tubes receiving reagents necessary for an amplification reaction and biological samples to be analyzed, designed to immobilize the tubes in a simple and safe manner even during a centrifugation operation while allowing the depositing operation inside the tubes to be secured.

[0011] To achieve such objectives, the object of the invention aims to propose a device comprising: a support tray for tubes receiving samples to be analyzed and into which the tip of a pipette is intended to be introduced, this tray comprising a plate having an upper face and a lower face and holes arranged according to a determined distribution opening into these two faces and adapted to each receive a tube resting on the upper surface and with its bottom projecting from the lower face of the tray, the plate of the tray being delimited by two longitudinal folded edges parallel to each other and connected to each other by two transverse folded edges parallel to each other;a cover comprising a plate having an upper face and a lower face, provided with chimneys arranged according to a distribution similar to the distribution of the holes in the tray and provided with stops to limit the insertion of pipette tips, the cover plate being delimited by two longitudinal folded edges parallel to each other and connected to each other by two transverse folded edges parallel to each other;the tray and the cover comprising a removable assembly system comprising two elastically deformable tabs arranged in the flanged edges of the cover and each cooperating with a complementary housing arranged in each of the flanged edges of the tray to produce an elastic fitting, when the cover occupies a superposed position above the tray and supported by its flanged edges on the tray to form a unitary assembly for holding the tubes and positioning the chimneys above the tubes to serve to guide the pipette tips penetrating inside the tubes and to stop them by the stops so that the distal end of the pipette tip comes substantially to the internal wall of the bottom of the tubes. ;

[0012] According to an advantageous feature, the cover is designed to occupy a superposed position below the tray for locating the chimneys so that they ensure the maintenance of the bottom of the tubes projecting from the underside of the tray.

[0013] In addition, the device according to the invention may further comprise in combination at least one and / or other of the following additional characteristics: the tray and the cover are made of translucent or transparent materials; the flanged edges of the cover delimit a border whose edge rests on the cover when the cover is in a superimposed position above the tray; the flanged edges of the cover and the flanged edges of the tray have interlocking parts intended to be interlocked together so that when the cover is assembled with the tray while being above the tray, the footprint of the cover delimited by its flanged edges is equal to the footprint of the tray delimited by its flanged edges; interlocking parts of the cover and the tray are provided with a foolproof system to ensure a direction of assembly of the cover on the tray;the cover comprises, from its upper face, a centering system for the tray resting by its fallen edges on the cover, the height of the fallen edges of the tray being adapted to ensure the engagement of the chimneys over a part of the height of the tubes from their bottom; the centering system is arranged set back from the fallen edges of the cover to receive the fallen edges of the tray so that the impression of the cover delimited by its fallen edges is equal to the impression of the tray delimited by its fallen edges; the chimneys of the cover comprise stops for limiting the insertion of the tips of the pipettes so that the tips come substantially up to the internal wall of the bottom of the tubes; the chimneys each rise projecting from the plate of the cover;the chimneys each delimit an opening having, from the upper face of the cover, a truncated cone-shaped section opening by its larger section, on the upper face of the cover to serve as a guide for the tip of the pipettes; the tray has support ribs projecting from the upper face of the tray to serve as a support for a support element of an analysis machine. ;

[0014] Another object of the invention is to propose a method of using a device according to the invention in the context of a process carrying out an amplification of nucleic acids.

[0015] The object of the invention is to propose a method of using a device in accordance with the invention, within the framework of a process carrying out an amplification of nucleic acids, using tubes intended to receive biological samples to be analyzed, the method consisting of: to insert into the holes of a tray, tubes pre-filled with a lyophilized or liquid reagent and each closed by a stopper provided with a slot allowing, by elastic deformation, the passage of a pipette tip, the stopper being supported on the upper surface of the tray; to place a cover in a superimposed position above and supported on the tray and to assemble it in a removable manner to the tray to form a unitary assembly for holding the tubes and with chimneys positioned above the tubes.

[0016] According to a preferred example of implementation, the method consists of: to introduce the tip of a liquid injection pipette through the chimneys of the lid which serve as a guide for the tip to penetrate through the stoppers and inside the tubes up to a stop carried by the chimneys of the lid so that the tip comes substantially up to the internal wall of the bottom of the tubes; to deliver the sample into the tubes using the pipette; and to remove the tip of the pipette from the tubes, stoppers and chimneys.

[0017] According to an alternative embodiment, a film capable of being perforated by a pipette tip is interposed between the stoppers and the cover, before assembly between the tray and the cover, so as to serve as an indicator of filling of the tubes with the aqueous solution.

[0018] According to another embodiment, a film capable of being perforated by a pipette tip is placed on the upper surface of the chimneys of the cover.

[0019] According to one step of the process, the unitary assembly for holding the tubes between the cover and the tray is introduced into a centrifugation machine, in particular a vertical one, to ensure a centrifugation operation of the contents of the tubes.

[0020] According to the invention, after carrying out the centrifugation operation: the cover is detached from the tray and placed on a support surface; the tray is placed in a superimposed position above the cover so as to engage the bottom of the tubes projecting from the lower face of the tray in the chimneys of the cover so that they hold the tubes; the tube caps are removed to be replaced by optical caps to allow an operation to be carried out for amplifying the nucleic acids contained in the tubes; placing the cover in a superimposed position above the tray and removably assembling it to the tray to form a unitary assembly for holding the tubes.

[0021] According to another step, the process consists of: loading the unitary tube holding assembly into an analysis machine; removing the cover from the tube support tray; holding the tubes against the tray, by a support element of the analysis machine, during a tube heating operation; and carrying out the operation of amplifying the contents of the tubes by the analysis machine. Brief description of the drawings

[0022] [ Fig. 1 ]There Figure 1 is a perspective view of the device according to the invention illustrated in a first configuration in which the tube support tray is located below a cover. Fig. 2 ]There Figure 2 is a perspective view showing the lower face of the tube support plate of the device according to the invention. Fig. 3 ]There Figure 3 is a perspective view showing the lower face of the cover of the device according to the invention. Fig. 4 ]There Figure 4 is a perspective of the device according to the invention illustrated in a first assembled position of superposition in which the cover is located above the tube support tray. Fig. 5 ]There Figure 5 is a sectional elevation view taken substantially along line V of the Figure 4 and showing a detail of the unitary assembly consisting of the assembly of the cover and the tube support plate. Fig. 6 ]There Figure 6 is a plan view of the device according to the invention illustrated in Figure 4 in the second assembled position of superposition and of which three pipette tips are introduced into tubes. Fig. 7 ]There Figure 7 is a sectional view of the device according to the invention in the second assembled superposition position, taken substantially along lines VII-VII of the Figure 6 . [ Fig. 8 ]There Figure 8 is a perspective view of the device according to the invention illustrated in a second configuration in which the cover is located below the tube support plate. Fig. 9 ]There Figure 9 is a perspective of the device according to the invention illustrated in a second assembled position of superposition in which the tube support tray is located above the cover. Fig. 10 ]There Figure 10 is a partly sectional perspective view of the device according to the invention illustrated in the second assembled position of superposition in which the tube support tray is located above the cover. Description of the embodiments

[0023] As can be seen from the drawings, the subject of the invention relates to a device 1 intended to be used advantageously in the context of a nucleic acid amplification process such as a Polymerase Chain Reaction or the like, using tubes 2 intended to receive biological samples to be analyzed and reagents necessary for an amplification reaction. Each tube 2 is pre-filled with a lyophilized reagent or a liquid reagent. By reagent is meant any substance or mixture of substances necessary for the implementation of an amplification reaction. Conventionally, the reagents are enzymes, nucleotides, primers, probes and other components necessary for amplification. In the case of PCR, a PCR mix is ​​used which contains the components necessary for PCR such as enzymes, nucleotides, ions, a buffer.

[0024] A sample is understood to be a small part or small quantity separated from an entity by a subtractive act usually called a sampling, for the purpose of analysis. The sample may be of human, animal, plant or environmental origin. It may correspond to a sample of biological fluid (whole blood, serum, plasma, urine, cerebrospinal fluid, organic secretion), a tissue sample or isolated cells. It may be of industrial origin, i.e., according to a non-exhaustive list, an air sample, a water sample, a sample taken from a surface, a part or a product being processed or manufactured, a product of food origin. This sample may possibly have undergone prior treatment, involving for example steps of lysis, mixing, dilution or even grinding, in particular if the starting entity is in the solid state. The sample deposited in the tubes may therefore be in liquid form.The biological sample analyzed is, in general, likely to - or suspected of - containing at least one target microorganism.

[0025] Each tube 2 has a tubular body 2t whose lower part is closed so as to have a bottom 2f from this closed lower end. Each tube 2 has an upper part 2s adapted to be equipped with a stopper 3 for closing the tube. This stopper 3 is provided with a slot 4 adapted to allow, by elastic deformation, the passage of a tip 5 of a pipette for injecting a liquid solution inside the tube 2.

[0026] According to the invention, the device 1 comprises a support plate 7 for the tubes 2 having an upper face 7s and a lower face 7i ( Figures 2 And 8 ). This tray 7 has holes 8 arranged to open into the upper face 7s and the lower face 7i. Each hole 8 is adapted to receive as illustrated in Figures 6, 7 And 10 , a tube 2 inserted inside the hole 8 and resting on the upper surface 7s of the plate while the bottom 2f of the tube projects from the lower face 7i of the plate 7 ( Figure 7 ).

[0027] The holes 8 are arranged in the plate 7 according to a determined distribution. Preferably, the holes 8 are arranged according to rows and columns. Typically, the plate 7 has eight rows of holes 8 referenced from A to H and twelve columns of holes 8 referenced from 1 to 12, i.e. 96 holes.

[0028] According to the invention, the device 1 also comprises a cover 10 having an upper face 10s and a lower face 10i, and provided with chimneys 11 internally delimiting openings 12 each opening onto the upper surface 10s and the lower surface 10i of the cover ( Figures 1 And 3). These chimneys 11 or openings 12 are arranged according to a distribution similar to the distribution of the holes 8 of the plate 7 so that in the position of superposition of the plate 7 and the cover 10, the holes 8 of the plate 7 coincide or are aligned with the openings 12 of the chimneys 11. In the example illustrated, the cover 10 comprises eight rows of chimneys 11 referenced from A to H and twelve columns of chimneys 11 referenced from 1 to 12, that is to say 96 chimneys 11 or openings 12.

[0029] According to the invention, the cover 10 and the tray 7 are adapted to cooperate together in a first stacking or superposition position and also, advantageously, in a second stacking or superposition position, each adapted for the implementation of specific steps in the process of analyzing the biological samples contained in the tubes. The cover 10 is designed to occupy at least one superposition position above the tray 7 so as to be removably assembled to the tray to form a unitary assembly for holding the tubes by the lower face of the cover and positioning the chimneys 11 above the tubes to serve for guiding the pipette tips ( Figures 1 , 4 , 5 , 6, 7). According to an advantageous embodiment characteristic, the cover 10 is designed to occupy a second position corresponding to a position of superposition of the cover 10 below the plate 7, in which the chimneys 11 are placed so as to ensure the maintenance of the bottom of the tubes projecting from the lower face 7i of the plate 7 ( Figures 8, 9 , 10 ).

[0030] According to a preferred embodiment, the tray 7 and the cover 10 have a rectangular shape. The tray 7 comprises a plate 7p delimited by two longitudinal flanged edges 7l parallel to each other and connected to each other by two transverse flanged edges 7t parallel to each other ( Figure 2 ). The plate 7p of the tray is thus provided along its entire periphery with the fallen edges 7l, 7t forming a continuous border extending in projection from the lower face 7i of the tray. The tray 7 is intended to be supported on a laying plane S by the edge of this continuous border so that advantageously, the height of the fallen edges 7t, 7l forming this border is substantially higher than the length of the tubes 2 projecting from the lower face 7i of the tray when the tubes 2 are supported by the tray ( Figure 7 ).

[0031] It should be noted that the lower face 7i of the plate 7 has a flat surface while the upper face 7s of the plate is provided with collars 7c each surrounding a hole 8 and allowing the positioning of the tubes 2 ( Figure 1 ). According to an advantageous embodiment characteristic, the plate 7 comprises support ribs 7n projecting from the upper face 7s of the plate to ensure homogeneous support for a support element of an analysis machine. As will be explained in detail in the remainder of the description, these support ribs 7n serve as angular compensation when the distribution of the tubes 2 is heterogeneous on the plate 7. As illustrated in Figure 8 , the support ribs 7n project from the upper face 7s of the plate near and parallel to the two transverse fallen edges 7t of the plate.

[0032] Similarly, the cover 10 comprises a plate 10p delimited by two longitudinal flanged edges 10l parallel to each other and connected to each other by two transverse flanged edges 10t parallel to each other ( Figures 1 And 3). The plate 10p of the cover 10 is thus provided along its entire periphery with the flanged edges 10l, 10t forming a border extending in projection from the lower face 10i of the cover. This border delimited by the flanged edges 10l, 10t has a rim 10r intended to come to bear on the upper face 7s of the plate 7 when the cover 10 occupies the superposition position above the plate 7. Thus, the cover 10 bears on the plate 7 by its flanged edges 10l, 10t. It should be noted that the lower face 10i of the cover has a flat surface while the upper face 10s of the cover is provided with the chimneys 11. In the example illustrated, the chimneys 11 rise in projection from the plate 10p of the cover. Of course, it can be provided that the chimneys 11 are made completely by the plate 10p of the cover so that the upper face 10s of the plate 10p would have a flat surface.In this embodiment, the plate 10p of the cover has a constant thickness and each chimney 11 is formed by the wall delimiting the opening 12 and whose height corresponds to the thickness of the plate 10p of the cover 10.

[0033] It must be considered that each chimney 11 delimits by its internal wall, a guide surface for the tip of a pipette during the operation of depositing a liquid inside the tube. Each opening 12 of a chimney 11 thus has a section and a shape adapted to ensure this guide function. As is apparent from the Figure 7 , each opening 12 of a chimney 11 has, from the upper face 10s of the cover, a truncated cone-shaped section 11a opening by its largest section, on the upper face 10s of the cover. For example, this truncated cone-shaped section 11a is extended by a tubular section 11b opening on the lower face 10i of the cover 10. According to an advantageous characteristic, the truncated cone-shaped section 11a of the chimneys is adapted to also ensure the engagement of the bottom of the tubes 2 which generally also has a truncated cone-shaped section ( Figure 10 ).

[0034] The chimneys 11 of the cover 10 have stops 11c for limiting the insertion of the tips 5 of the pipettes so that the tips come substantially up to the internal wall of the bottom of the tubes ( Figure 7 ). In other words, the stops 11c are arranged on the cover 10 so that in the position in which the cover 10 is resting on the plate 7, the distal end of the tip 5 of the pipette arrives, at the end of the engagement stroke, close to the wall of the bottom of the tube as is apparent from the Figure 7 The position of the stops 11c relative to the support rim 10r is such that in the position of superposition of the cover 10 above the plate 7, the distal end of the tip 5 of the pipette is located close to the bottom wall of the tube. For example, the stops 11c are formed by the upper rim of the chimneys 11 on which come into contact fins 5a of the tips 5 of the pipettes. In this stop or stop position, the distal end of the tip 5 of the pipette is located close to the bottom wall of the tube, when the cover 10 occupies its first superposition position resting on the plate 7. The positioning of the distal end of the tip 5 of the pipette close to the bottom wall of the tube thus makes it possible to secure the operation of depositing the liquid inside the tube 2.Indeed, when depositing a small volume, it is necessary to make the drop touch the internal wall of the tube in order to prevent it from remaining attached to the tip of the pipette. Its transfer is thus carried out reliably according to good practices for using a pipette.

[0035] According to a preferred embodiment, the tray 7 and the cover 10 are made of translucent or transparent materials. Typically, the tray 7 and the cover 10 are generally made of polycarbonate, but could also be made of polypropylene, high-density polyethylene.

[0036] According to a characteristic of the invention, the tray 7 and the cover 10 comprise a removable assembly system 13 allowing the cover 10 and the tray 7 to be secured together to form a unitary assembly for holding the tubes 2. In this assembled position, the cover 10 occupies a superposition position above the tray 7, with the cover 10 which is supported on the tray 7 and with the plate 10p of the cover located above the plate 7p of the tray. The removable assembly system 10 is also adapted so that the cover 10 and the tray 7 can be easily separated so that they can be handled independently and positioned for example to occupy another stacked position. The removable assembly system 13 can be produced in any suitable manner. In the preferred embodiment illustrated in the drawings, the removable assembly system 13 is of the elastic fitting type.

[0037] According to the embodiment illustrated in the drawings, the tray 7 and the cover 10 comprise, as a removable assembly system 13, two tabs 13a arranged in the preferably transverse flanged edges 10t of the cover 10 and each cooperating with a complementary housing 14 arranged in each of the transverse flanged edges 7t of the tray 7 to produce an elastic fitting. As is apparent from Figures 1 , 3 , 4 et 5 , each transverse fallen edge 10t of the cover 10 comprises two cutouts 10tf delimiting between them, an elastically deformable tab 13a. Each tab 13a is provided at its free end, on its internal surface, with a rib 13b intended to engage in the complementary housing 14 which delimits a stop surface 14a for the rib 13b ( Figure 5 ). When the legs 13a of the cover 10 are blocked by the stop surfaces 14a of the plate 7, the lower face of the plate 10p of the cover is positioned above and close to or in contact with the caps of the tubes 2 supported by the plate 7. In this position, the tubes 2 are immobilized, between the plate 7 and the cover 10.

[0038] According to an advantageous embodiment characteristic, the flanged edges 10t, 10l of the cover 10 and the flanged edges 7t, 7l of the plate 7 comprise nesting parts respectively 10tm, 10tf, 10lm, 10lf and 7tm, 7tf, 7lm, 7lf intended to be nested together so that, when the cover 10 is assembled with the plate 7 while being above the plate 7, the footprint of the cover 10 delimited by its flanged edges 10t, 10l is equal to the footprint of the plate 7 delimited by its flanged edges 7t, 7l. In this position illustrated in Figure 4 , the unitary assembly constituted by the cover 10 and the plate 7 is a parallelepiped solid with flat lateral surfaces formed by the fallen edges 10t, 10l of the cover aligned with the fallen edges 7t, 7l of the plate.

[0039] The interlocking between the flanged edges 10t, 10l of the cover 10 and the flanged edges 7t, 7l of the plate 7 can be obtained using the interlocking parts 10tm, 10tf, 10lm, 10lf and 7tm, 7tf, 7lm, 7lf produced in any suitable manner. In the embodiment illustrated in the Figures, each longitudinal flanged edge 7l of the plate 7 comprises two interlocking parts 7lf called female formed by housings arranged in the longitudinal flanged edges 7l and produced on either side of a interlocking part called male 7lm. Each longitudinal fallen edge 10l of the cover 10 comprises two interlocking parts 10lm called male parts delimited on either side by an interlocking part called female part 10lf made by a cutout.The two male interlocking parts 10lm and the female interlocking part 10lf of each longitudinal flanged edge 10l of the cover 10 are intended to interlock respectively in the two female interlocking parts 7lf and in the male interlocking part 7lm of each transverse edge 7t of the plate 7. Of course, the female interlocking parts 10lf and the male interlocking parts 10lm of the cover have congruent or complementary shapes respectively to the male interlocking parts 7lm and to the female interlocking parts 7lf of the plate 7.

[0040] Similarly, each transverse fallen edge 10t of the cover 10 comprises two so-called female fitting parts 10tf formed by the cutouts made on either side of a male fitting part made by the tab 13a, these cutouts 15 leaving a male fitting part 10tm remaining up to each corner. The two female nesting parts 10tf and the three male nesting parts 13a, 10tm of each transverse flanged edge 10t of the cover 10 are intended to nest respectively in two male nesting parts 7tm and in three female nesting parts 7tf of each transverse flanged edge 7t of the plate 7. The three female nesting parts 7tf of each transverse flanged edge 7t of the plate 7 are for example housings arranged in these transverse flanged edges 7t of the plate 7.Of course, the female interlocking parts 10tf and the male interlocking parts 13a, 10tm of the cover 10 have congruent or complementary shapes respectively to the male interlocking parts 7tm and to the female interlocking parts 7tf of the plate 7. It should be noted that the male interlocking parts 10lm of the longitudinal flanged edges and the male interlocking parts 13a, 10tm of the transverse flanged edges 10t of the cover 10 extend projecting from the rim 10r of the flanged edges and over only part of the width of this border so that the rim 10r remains at the right of the male interlocking parts (. Figure 3 ). The cover 10 thus has, by its fallen edges, a rim 10r extending substantially continuously over the entire periphery of the plate 10p, making it possible to obtain stable support for the cover 10 on the plate 7.

[0041] According to an advantageous embodiment feature, the interlocking parts of the cover 10 and the plate 7 are provided with a foolproof system to ensure a direction of mounting of the cover on the plate. According to the embodiment example illustrated in the Figures, the two female interlocking parts 7lf of a longitudinal fallen edge of the plate 7 and consequently the two corresponding male interlocking parts 10lm of the cover are arranged asymmetrically. As is apparent for example from the Figure 1 , the length taken along a longitudinal edge of a female fitting part 7lf of the plate (respectively of a male fitting part 10lm) is different from the length of the other female fitting part 7lf of the plate (respectively of the other male fitting part 10lm). Such an asymmetrical embodiment of the fitting parts allows the cover 10 to be mounted on the support 7 in a single direction.

[0042] As previously indicated, the cover 10 is designed to occupy a second stacking position corresponding to a superposition position of the cover 10 below the tray 7 ( Figures 8, 9 And 10 ). In this second superposition position, the plate 7 rests on the upper face 10s of the cover 10, by the edge of its fallen edges 7t, 7l. According to a preferred embodiment characteristic, the cover 10 comprises, from its upper face 10s, a centering system 17 for the plate 7 so that the cover 10 and the plate 7 are secured together at least in a horizontal plane.

[0043] According to a preferred embodiment, the centering system 17 is arranged so that the footprint of the cover 10 delimited by its flanged edges 10t, 10l is equal to the footprint of the plate 7 delimited by its flanged edges 7t, 7l. In other words, the longitudinal 10l and transverse 10t flanged edges of the cover 10 are located vertically or in the extension respectively of the longitudinal 7l and transverse 7t flanged edges of the plate. As is more precisely shown in the Figure 8 , the centering system 17 is arranged set back from the fallen edges of the cover 10 to receive the fallen edges of the plate 7 so that the fallen edges 10t, 10l of the cover 10 are aligned with the fallen edges 7t, 7l of the plate 7.

[0044] In the example illustrated, the centering system 17 comprises a groove 17a arranged from the upper face 10s of the cover 10, opening at the periphery of the plate 10p of the cover to allow the engagement of the longitudinal 7l and transverse 7t flanged edges of the plate. This groove 17a thus delimits a centering rim 17b for the longitudinal 7l and transverse 7t flanged edges of the plate. For example, this centering rim 17b may be extended punctually, at each corner of the cover 10, by centering studs 17c projecting from the upper face of the cover 10. It should be noted that it may be provided that the centering rim 17b of the cover 10 has a section adapted to produce, with the section delimited by the internal surface of the flanged edges of the plate 7, a tight fit leading to complete attachment of the plate 7 to the cover 10.The tray 7 and the cover 10 thus form a unitary assembly in this second stacking position.

[0045] In this second stacking position, the height of the dropped edges 7t, 7l of the tray 7 is adapted to ensure the engagement of the chimneys 11 of the cover over part of the height of the tubes 2 from their bottom ( Figure 10 ). The engagement of the tubes 2 by their bottom inside the openings 12 of the chimneys 11 ensures centering of the tubes preventing them from tilting, in particular when carrying out various interventions on the tubes and / or the plugs, as will be explained in detail in the rest of the description.

[0046] It is clear from the above description that the device 1 according to the invention comprises two separate elements which can be secured together in two different stacking positions to allow the tubes to be handled in order to carry out various steps in the treatment of the biological samples contained in these tubes. It should be noted that such devices 1 can be stacked or stacked on top of each other in either superposition position.

[0047] The device 1 according to the invention is particularly suitable for use in the context of a process carrying out amplification of nucleic acids such as the Polymerase Chain Reaction or the like, using tubes 2 intended to receive biological samples to be analyzed. Each tube 2 may be pre-filled with a lyophilized or liquid reagent. It should be noted that the contents of each tube 2 may also have undergone one or more treatment steps, such as a lyophilization operation. Each tube 2 thus pre-filled is closed using a stopper 3 provided with a slot 4 allowing, by elastic deformation, the passage of a tip 5 of a pipette.

[0048] The method of implementing device 1 consists of: to insert into the holes 8 of the tray 7, the tubes 2 pre-filled with a reagent and each closed by a stopper 3 provided with a slot and resting on the upper surface 7s of the tray 7; to place the cover 10 in a superposition position above the tray 7 ( Figure 1 ) and to removably assemble the cover 10 to the tray 7 to form a unitary assembly ( Figure 4 ).

[0049] The assembly of the cover 10 to the plate 7 is carried out in a simple manner by bringing the cover 10 closer to the plate 7. During this bringing together, the elastic tabs 13a are elastically deformed outwards until the ribs 13b of the elastic tabs 13a penetrate into the housings 14 to be blocked by the stop surfaces 14a of the plate 7. It should be noted that during this assembly operation, the cover 10 is guided in sliding on the plate 7 by the cooperation of the interlocking parts of the cover 10 with the interlocking parts of the plate 7.

[0050] In this assembly position, the lower face 10i of the plate 10p of the cover 10 is positioned above the caps of the tubes 2 supported by the plate 7 to hold them in position. The cover 10 rests on the plate 7, by the rim 10r of its fallen edges 10l, 10t. In this position, the tubes 2 are immobilized or blocked, between the plate 7 and the cover 10 and the chimneys 11 of the cover 10 are positioned above the tubes 2 ( Figure 7 ).

[0051] The next step in preparing a sample for a Polymerase Chain Reaction or the like is to insert the tip 5 of a liquid injection pipette through the chimneys 11 of the lid and the slots 4 of the stoppers 3 so as to open inside the tubes 2 ( Figures 6 et 7 ). It must first be noted that the chimneys 11 guide the operator to correctly position the tip of the pipette to ensure its introduction inside the tubes 2. In fact, the operator introduces into the opening 12 of the chimneys 11, the tip 5 which is guided throughout its travel, namely when approaching the stopper 3 by the truncated cone-shaped portion 11a, during its penetration into the slot 4 of the stopper which deforms elastically, and inside the tube 2.

[0052] Advantageously, the stroke of the pipette is limited by the stops 11c of the chimneys 11 so that the distal end of the tip of the pipette is close to, or even in contact with, the bottom wall of the tube. The tip of the pipette is ideally positioned close to the wall of the tube so as to ensure the operation of depositing the liquid inside the tube 2. This depositing operation, which is a delicate operation to carry out successfully since the liquid to be deposited is generally a drop of small volume, typically 5 to 10 microliters, is thus made safe.

[0053] After delivering the liquid into the tubes 2 using the pipette, the operator removes the tip from the pipette, tubes 2, caps 3 and chimneys 11. The delivered liquid is the sample. A reagent necessary for amplification can also be added.

[0054] It should be noted that the transparency of the tray 7 and the cover 10 gives the operator the possibility of seeing the color code inside the tubes 2.

[0055] According to an alternative embodiment, it should be noted that a film capable of being perforated by a pipette tip is placed on the cover 10 so as to serve as an indicator of the filling of the tubes 2 with the liquid. This film, not shown in the drawings, can be placed on the upper surface of the chimneys 11 of the cover 10.

[0056] According to another embodiment, before assembly between the tray 7 and the cover 10, a film capable of being perforated by a pipette tip can be interposed between the stoppers 3 and the cover 10, so as to serve as an indicator of filling of the tubes 2 with the liquid. This film, not shown in the drawings, can be deposited directly on the upper surface 7s of the tray provided with the stoppers 3.

[0057] This unitary assembly with the tubes 2 immobilized between the cover 10 and the tray 7 can be introduced into a centrifugation machine of any type known per se to ensure a centrifugation operation of the contents of the tubes 2. According to an advantage of the device 1 according to the invention, the device 1 can be loaded into a vertical centrifugation machine without risk of leaks from the tubes. The compact nature of the device 1 facilitates the loading and unloading operations of the device 1.

[0058] After the centrifugation operation has been carried out, the contents of the tubes 2 are intended to undergo an amplification reaction of the nucleic acids contained in the tubes using an analysis machine of any type known per se, typically a thermocycler. According to an alternative implementation, the caps 3 closing the tubes 2 are replaced by optical caps before the operation of analyzing the contents of the tubes.

[0059] For this purpose, the cover 10 is detached from the plate 7 and placed on a laying plane S. This operation is relatively easy to carry out since it is sufficient to spread the elastic tabs 13a to release the ribs 13b from the housings 14 and to lift, in a translational movement, the cover 10 relative to the plate 7. After having placed the cover 10 in support on a laying plane S, by its flanged edges 10l, 10t, the plate 7 is placed in a superposition position above the cover 10 so as to engage the bottom of the tubes projecting from the lower face of the plate in the chimneys 11 of the cover so that they ensure the holding of the tubes. As this emerges more precisely from the Figure 10, each bottom of the tubes 2 is engaged in the opening 12 of the chimneys 11 avoiding any tilting movement during the operation of removing the stoppers replaced by optical stoppers for carrying out the operation of analyzing the contents of the tubes. It should be noted that the operation of analyzing the contents of the tubes can be carried out without changing the stoppers 3 provided with a slot and initially closing the tubes.

[0060] To carry out the amplification of the nucleic acids contained in the tubes 2, the unitary assembly consisting of the tray 7 for supporting the tubes 2 assembled with the cover 10 is loaded into the analysis machine. In the case where the caps have been replaced by optical caps, the tray 7 is released from the cover 10 so that the cover 10 can be placed above the tray 7 and secured together using the removable assembly system 13 as already explained. This unitary assembly makes it possible to easily and safely load all the tubes 2 into the analysis machine.

[0061] The cover 10 of the device 1 is removed from the tray 7 so as to allow the heating of the tubes 2 and the analysis operation. For this purpose, a support element of the analysis machine such as its cover holds the tubes 2 against the tray 7 while ensuring good contact of the tubes with the heating block of the analysis machine and in particular with the tube receiving wells arranged in the heating block. Advantageously, the cover of the analysis machine comes into contact with the support ribs 7n of the tray 7. The operation of amplifying the nucleic acids contained in the tubes 2 can therefore be carried out by the analysis machine.

[0062] It should be noted that during the amplification cycle, the tubes 2 are clamped in the heating block by the support element of the analysis machine. At the end of the cycle, it is possible to unload the tubes 2 from the analysis machine using the tray 7. This allows all the tubes 2 having undergone the reaction to be removed simultaneously and easily.

Claims

1. Device comprising: - a support tray (7) for tubes (2) receiving samples to be analysed and into which the tip (5) of a pipette is intended to be inserted, this tray (7) including a plate (7p) having an upper face (7s) and a lower face (7i) and holes (8) arranged in a predetermined pattern, opening into these two faces and each designed to receive a tube bearing against the upper surface and with the bottom thereof projecting from the lower face of the tray, the plate (7p) of the tray being delimited by two longitudinal flanged edges (71) parallel to each other and connected to each other by two transverse flanged edges (7t) parallel to each other; - a cover (10) comprising a plate (10p) having an upper face (10s) and a lower face (10i), provided with chimneys (11) arranged in a pattern similar to the pattern of the holes in the tray and provided with stops (11c) limiting the insertion of the pipette tips (5), the plate (10p) of the cover being delimited by two longitudinal flanged edges (101) parallel to each other and connected to each other by two transverse flanged edges (10t) parallel to each other; - the tray (7) and the cover (10) including a removable assembly system (13) including two elastically deformable tabs (13a) formed in the flanged edges of the cover and each cooperating with a complementary seat (14) formed in each of the flanged edges of the tray to achieve elastic interlocking, when the cover (10) is in a stacking position above the tray (7) and the flanged edges thereof are bearing against the tray to form a unitary assembly for holding the tubes and to position the chimneys (11) above the tubes to guide the pipette tips penetrating the inside of the tubes and to stop them by means of the stops (11c) so that the distal end of the tip of the pipette substantially reaches the inner wall of the bottom of the tubes.

2. Device according to Claim 1, wherein the cover (10) is designed to be in a stacking position beneath the tray (7) to arrange the chimneys (11) such that they hold the bottom of the tubes projecting from the lower face of the tray.

3. Device according to one of Claims 1 and 2, wherein the tray (7) and the cover (10) are made of translucent or transparent materials.

4. Device according to one of Claims 1 to 3, wherein the flanged edges (101, 10t) of the cover (10) delimit an edge, the rim (10r) of which bears against the cover (10) when the cover (10) is in a stacking position above the tray (7).

5. Device according to one of Claims 1 to 4, wherein the flanged edges of the cover (10) and the flanged edges of the tray (7) have interlocking portions (10tm, 10tf, 10lm, 10lf and 7tm, 7tf, 7lm, 7lf) intended to be interlocked together so that, when the cover (10) is joined to the tray (7) from above the tray (7), the footprint of the cover (10) delimited by the flanged edges (10t, 101) thereof is identical to the footprint of the tray (7) delimited by the flanged edges (71, 7t) thereof.

6. Device according to the preceding claim, wherein interlocking portions (10tm, 10tf, 10lm, 10lf and 7tm, 7tf, 7lm, 7lf) of the cover (10) and of the tray (7) are provided with a mistake-proofing system to guarantee a mounting direction of the cover (10) on the tray (7).

7. Device according to one of Claims 1 to 6, wherein the cover (10) includes, from the upper face (10s) thereof, a centring system (17) for the tray (7) bearing against the cover (10) via the flanged edges thereof, the height of the flanged edges (71, 7t) of the tray being designed to engage the chimneys (11) over a portion of the height of the tubes from the bottom thereof.

8. Device according to the preceding claim, wherein the centring system (17) is set back from the flanged edges (101, 10t) of the cover (10) to receive the flanged edges of the tray so that the footprint of the cover delimited by the flanged edges thereof is identical to the footprint of the tray delimited by the flanged edges thereof.

9. Device according to one of the preceding claims, according to which the chimneys (11) of the cover (10) have stops (11c) for limiting the insertion of the tips of the pipettes so that the tips substantially reach the inner wall of the bottom of the tubes.

10. Device according to one of Claims 1 to 9, wherein each chimney (11) projects from the plate (10p) of the cover (10).

11. Device according to one of Claims 1 to 10, wherein each chimney (11) delimits an opening (12) having, from the upper face (10s) of the cover, a frustoconical section (11a), the largest section of which opens into the upper face (10s) of the cover to guide the tips of the pipettes.

12. Device according to one of the preceding claims, wherein the tray (7) has bearing ribs (7n) projecting from the upper face (7s) of the tray to bear a bearing element of an analysis machine.

13. Method of using a device according to one of Claims 1 to 12, in the context of a nucleic acid amplification process, using tubes (2) intended to receive biological samples to be analysed, the method consisting of: - inserting, into the holes (8) of a tray (7), tubes pre-filled with a liquid or freeze-dried reagent and each closed by a cap (3) provided with a slot (4) allowing a tip (5) of a pipette to pass through by elastic deformation, the cap bearing against the upper surface of the tray; - placing a cover (10) in a stacking position above and bearing against the tray (7) and joining it removably to the tray to form a unitary assembly for holding the tubes, with chimneys (11) positioned above the tubes.

14. Method according to the preceding claim, consisting of: - inserting the tip (5) of a pipette for injecting a liquid through the chimneys (11) of the cover which guide the tip to penetrate through the caps (3) and into the tubes (2) as far as a stop (11c) carried by the chimneys (11) of the cover (10) so that the tip substantially reaches the inner wall of the bottom of the tubes; - delivering the sample into the tubes using the pipette; and - removing the tip (5) of the pipette from the tubes (2), the caps (3) and the chimneys (11).

15. Method according to one of Claims 13 and 14, wherein a film capable of being perforated by a tip of a pipette (5) is interposed between the caps (3) and the cover, before the tray is joined to the cover, to act as an indicator of the filling of the tubes with the liquid.

16. Method according to one of Claims 13 and 14, wherein a film capable of being perforated by a tip of a pipette (5) is placed on the upper surface of the chimneys (11) of the cover (10).

17. Method according to one of Claims 13 to 16, wherein the unitary assembly for holding the tubes between the cover (10) and the tray (7) is inserted into a centrifuge machine, notably a vertical centrifuge machine, to centrifuge the content of the tubes.

18. Method according to the preceding claim, wherein, after centrifuging has been carried out: - the cover (10) is detached from the tray (7) and placed on a standing plane; - the tray (7) is placed in a stacking position above the cover (10) so as to engage the bottom of the tubes projecting from the lower face of the tray in the chimneys (11) of the cover such that they hold the tubes; - the caps (3) of the tubes are removed and replaced by optical caps to enable an amplification operation of the nucleic acids contained in the tubes to be carried out; - placing the cover (10) in a stacking position above the tray (7) and joining it removably to the tray to form a unitary assembly for holding the tubes.

19. Method according to one of Claims 17 and 18, wherein the method consists of: - loading the unitary tube-holding assembly into an analysis machine; - removing the cover (10) from the tube-support tray (7); - holding the tubes against the tray, using a bearing element of the analysis machine, during an operation to heat the tubes; and - carrying out the amplification operation on the content of the tubes using the analysis machine.

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