Patent Application
20230211348
The present invention relates to the technical field of devices used to handle tubes receiving biological samples to be analyzed.
The device in accordance with the invention generally relates to the process of samples for analysis, in the field of microbiology. It finds particularly advantageous applications to help in the handling of the tubes receiving biological samples to be analyzed by nucleic acid amplification techniques such as the Polymerase Chain Reaction (or PCR) and similar methods.
The nucleic acid amplification techniques are well-known techniques in the prior art. These techniques and in particular the PCR implement different steps to prepare and analyze biological samples, in particular steps of rehydration of reagents, mixing of reagents with the sample, centrifugation, amplification and detection. This technique uses tubes pre-filled with reagents and each closed with a sealing plug. These tubes are intended to receive the biological samples to be analyzed. These tubes are handled by suitable devices in order to carry out the different steps of this technique.
In the prior art, there are many devices to ensure the implementation of the PCR technique while allowing the simple and safe handling of the tubes when carrying out the different steps of this technique.
For example, document CN 107868752 describes a device including a support plate for sample tubes to be analyzed, intended to be supported by a stand provided with a positioning base in an analysis machine. This device also includes a lid provided with legs intended to be magnetically fixed to the stand, so that the tube support plate is immobilized between the stand and the lid. Moreover, this lid is provided with passage holes for a pipette tip that are matching the tubes. Such a device allows simultaneously handling a set of tubes for its loading and its positioning in the analysis machine.
However, such a device does not guarantee the immobilization of the tubes on the stand. Indeed, the system for assembling the lid to the stand by a magnetic force does not allow developping a sufficient assembly force when the tubes supported by such a device undergo in particular the centrifugation operation. Moreover, the presence of the lid above the tubes makes the operation of deposition inside the tubes even more complex, operation which is delicate to conduct since the pipette must ensure the deposition of a drop of small volume, typically from 5 to 10 microliters, inside the tubes.
U.S. Pat. No. 7,329,393 describes a device for supporting sample tubes to be analyzed, adapted to undergo a centrifugation operation. Such a device includes a lid intended to be fixed to a base by a screw assembly system and between which a tube receiving tray and a support block are interposed. If such a device allows an effective holding of the tubes when they undergo the centrifugation operation, the mounting and dismounting operations prove to be relatively long to conduct. It follows that the use of such a device is limited in practice to this centrifugation operation. Moreover, such a device does not provide a solution to the problem of deposition of the samples inside the tubes.
The patent application US 2015/0132841 describes an assembly including a support for containers receiving samples to be analyzed and a lid provided with openings arranged according to a distribution similar to the containers. The lid is temporarily assembled to the support using fixing elements such as elastically deformable arms extending from the support and cooperating with protrusions carried by the lid. Such an assembly allows holding the containers in position. However, such an assembly does not provide a solution to the problem of deposition of the samples inside the containers.
One object of the invention is to overcome 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 able to be implemented in a simple and effective manner for different prior operations or during an amplification reaction.
One 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 a securing of the operation of deposition inside the tubes.
To achieve such objectives, the object of the invention aims to propose a device comprising:
According to one advantageous characteristic, the lid is designed to occupy a position of superposition below the tray to place the chimneys so that they ensure the holding of the bottom of the tubes protruding from the lower face of the tray.
In addition, the device according to the invention can further include in combination at least either or both of the following additional characteristics:
Another object of the invention is to propose a method for using a device in accordance with the invention as part of a process performing a nucleic acid amplification.
The object of the invention aims to propose a method for using a device in accordance with the invention, as part of a process performing a nucleic acid amplification, implementing tubes intended to receive biological samples to be analyzed, the method consisting of:
According to one preferred exemplary implementation, the method consists of:
According to one variant, a film able to be perforated by a tip of a pipette is interposed between the plugs and the lid, before the assembly between the tray and the lid, so as to serve as a fill indicator for filling the tubes with the aqueous solution.
According to another variant, a film able to be perforated by a tip of a pipette is placed on the upper surface of the chimneys of the lid.
According to one step of the method, the unitary assembly for holding the tubes between the lid and the tray is introduced into a centrifugation machine, in particular a vertical machine, to ensure an operation of centrifugation of the contents of the tubes.
According to the invention, after carrying out the centrifugation operation:
According to another step, the method consists of:
As appearing from the drawings, the object of the invention relates to a device 1 intended to be used advantageously as part of a nucleic acid amplification process such as a Polymerase Chain Reaction or the like, implementing 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, it 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 the amplification. In the case of the PCR, a PCR mix which contains the components necessary for the PCR such as enzymes, nucleotides, ions, a buffer, is used.
By sample, it is meant a small part or small amount separated from an entity by a subtractive act usually called collection, for the purpose of analysis. The sample can be of human, animal, plant or environmental origin. It can correspond to a biological fluid collection (whole blood, serum, plasma, urine, cerebrospinal fluid, organic secretion), a tissue collection or isolated cells. It can be of industrial origin, i.e., according to a non-exhaustive list, an air collection, a water collection, a collection taken from a surface, a part or a product being treated or manufactured, a product of food origin. This sample may possibly have undergone a prior treatment, involving for example lysis, mixing, dilution or grinding steps, particularly if the originating entity is in the solid state. The sample deposited in the tubes can therefore be in liquid form. The biological sample analyzed is, in general, likely to contain or suspected of containing at least one target microorganism.
Each tube 2 has a tubular body 2t whose lower part is closed so as to present a bottom 2f from this closed lower end. Each tube 2 has an upper part 2s adapted to be equipped with a plug 3 for sealing the tube. This plug 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.
In accordance with the invention, the device 1 comprises a support tray 7 for the tubes 2 having an upper face 7s and a lower face 7i (
The holes 8 are arranged in the tray 7 according to a determined distribution. Preferably, the holes 8 are arranged in rows and columns. Typically, the tray 7 includes eight rows of holes 8 referenced from A to H and twelve columns of holes 8 referenced from 1 to 12, that is to say 96 holes.
In accordance with the invention, the device 1 also comprises a lid 10 having an upper face 10s and a lower face 10i, and provided with chimneys 11 internally delimiting openings 12 each opening out onto the upper surface 10s and the lower surface 10i of the lid (
In accordance with the invention, the lid 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 lid 10 is designed to occupy at least one position of superposition 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 lid and to position the chimneys 11 above the tubes to guide the pipette tips (
According to one preferred variant, the tray 7 and the lid 10 have a rectangular shape. The tray 7 includes 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 (
It should be noted that the lower face 7i of the tray 7 has a planar surface while the upper face 7s of the tray is provided with flanges 7c each surrounding a hole 8 and allowing the positioning of the tubes 2 (
Similarly, the lid 10 includes 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 (
It must be considered that each chimney 11 delimits by its inner wall, a guide surface for the tip of a pipette during the operation of deposition of a liquid inside the tube. Each opening 12 of a chimney 11 thus has a section and a shape adapted to ensure this guiding function. As seen in
The chimneys 11 of the lid 10 include abutments 11c for limiting the depression of the tips 5 of the pipettes so that the tips substantially reach the inner wall of the bottom of the tubes (
According to one preferred variant, the tray 7 and the lid 10 are made of translucent or transparent materials. Typically, the tray 7 and the lid 10 are generally made of polycarbonate, but could also be made of polypropylene, high—density polyethylene.
According to one characteristic of the invention, the tray 7 and the lid 10 include a removable assembly system 13 allowing the lid 10 and the tray 7 to be fastened together to form a unitary assembly for holding the tubes 2. In this assembled position, the lid 10 occupies a position of superposition above the tray 7, with the lid 10 bearing on the tray 7 and with the plate 10p of the lid located above the plate 7p of the tray. The removable assembly system 10 is also adapted so that the lid 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 made in any suitable manner. In the preferred exemplary embodiment illustrated in the drawings, the removable assembly system 13 is of the elastic interlocking type.
According to the exemplary embodiment illustrated in the drawings, the tray 7 and the lid 10 include, as a removable assembly system 13, two legs 13a arranged in the preferably transverse flanged edges 10t of the lid 10 and each cooperating with a complementary housing 14 arranged in each of the transverse flanged edges 7t of the tray 7 to achieve an elastic interlocking. As seen in
According to one advantageous characteristic of embodiment, the flanged edges 10t, 10l of the lid 10 and the flanged edges 7t, 7l of the tray 7 include nesting parts respectively 10tm, 10tf, 10lm, 10lf and 7tm, 7tf, 7lm, 7lf intended to be nested together so that, when the lid 10 is assembled with the tray 7 while being above the tray 7, the footprint of the lid 10 delimited by its flanged edges 10t, 10l is equal to the footprint of the tray 7 delimited by its flanged edges 7t, 7l. In this position illustrated in
The nesting between the flanged edges 10t, 10l of the lid 10 and the flanged edges 7t, 7l of the tray 7 can be obtained using the nesting parts 10tm, 10tf, 10lm, 10lf and 7tm, 7tf, 7lm, 7lf made in any suitable manner. In the exemplary embodiment illustrated in the Figures, each longitudinal flanged edge 7l of the tray 7 includes two nesting parts 7lf called female nesting parts formed by housings arranged in the longitudinal flanged edges 7l and made on either side of a nesting part called male nesting part 7lm. Each longitudinal flanged edge 10l of the lid 10 includes two nesting parts 10lm called male nesting parts delimited on either side by a nesting part called female nesting part 10lf made by a cutout. The two male nesting parts 10lm and the female nesting part 10lf of each longitudinal flanged edge 10l of the lid 10 are intended to be nested respectively into the two female nesting parts 7lf and in the male nesting part 7lm of each transverse edge 7t of the tray 7. Of course, the female nesting parts 10lf and the male nesting parts 10lm of the lid have shapes respectively congruent or complementary to the male nesting parts 7lm and to the female nesting parts 7lf of the tray 7.
Similarly, each transverse flanged edge 10t of the lid 10 includes two nesting parts called female nesting parts 10tf formed by the cutouts made on either side of a male nesting part made by the leg 13a, these cutouts 15 leaving to each corner a male nesting part 10tm. The two female nesting parts 10tf and the three male nesting parts 13a, 10tm of each transverse flanged edge 10t of the lid are intended to be nested respectively into two male nesting parts 7tm and in three female nesting parts 7tf of each transverse flanged edge 7t of the tray 7. The three female nesting parts 7tf of each transverse flanged edge 7t of the tray 7 are for example housings arranged in these transverse flanged edges 7t of the tray 7. Of course, the female nesting parts 10tf and the male nesting parts 13a, 10tm of the lid 10 have shapes respectively congruent or complementary to the male nesting parts 7tm and to the female nesting parts 7tf of the tray 7. It should be noted that the male nesting parts 10lm of the longitudinal flanged edges and the male nesting parts 13a, 10tm of the transverse flanged edges 10t of the lid 10 protrude from the rim 10r of the flanged edges and over only part of the width of this border so that the rim 10r remains in line with the male nesting parts (
According to one advantageous characteristic of embodiment, the nesting parts of the lid 10 and the tray 7 are arranged with a foolproof system to ensure a direction of mounting of the lid on the tray. According to the exemplary embodiment illustrated in the Figures, the two female nesting parts 7lf of a longitudinal flanged edge of the tray 7 and consequently the corresponding two male nesting parts 10lm of the lid are arranged asymmetrically. As is apparent for example from
As indicated above, the lid 10 is designed to occupy a second stacking position corresponding to a position of superposition of the lid 10 below the tray 7 (
According to one preferred variant, the centering system 17 is arranged so that the footprint of the lid 10 delimited by its flanged edges 10t, 10l is equal to the footprint of the tray 7 delimited by its flanged edges 7t, 7l. In other words, the longitudinal 10l and transverse 10t flanged edges of the lid 10 are located in alignment with or in the extension of respectively the longitudinal 7l and transverse 7t flanged edges of the tray. As seen more specifically in
In the example illustrated, the centering system 17 includes a groove 17a arranged from the upper face 10s of the lid 10, by opening at the periphery of the plate 10p of the lid to allow the engagement of the longitudinal 7l and transverse 7t flanged edges of the tray. This groove 17a thus delimits a centering rim 17b for the longitudinal 7l and transverse 7t flanged edges of the tray. For example, this centering rim 17b can be extended punctually, at each corner of the lid 10, by centering studs 17c protruding from the upper face of the lid 10. It should be noted that it can be provided that the centering rim 17b of the lid 10 has a section adapted to achieve, with the section delimited by the inner surface of the flanged edges of the tray 7, a tight fit leading to a complete fastening of the tray 7 with the lid 10. The tray 7 and the lid 10 thus form a unitary assembly in this second stacking position.
In this second stacking position, the height of the flanged edges 7t, 7l of the tray 7 is adapted to ensure the engagement of the chimneys 11 of the lid over part of the height of the tubes 2 from their bottom (
It appears from the foregoing description that the device 1 in accordance with the invention includes two separate elements which can be fastened together in two different stacking positions to allow the handling of the tubes in order to ensure the accomplishment of various steps of treating the biological samples contained in these tubes. It should be noted that such devices 1 can be piled or stacked on top of each other in either of the superposition position.
The device 1 in accordance with the invention is particularly suitable for use as part of a process performing a nucleic acid amplification such as the Polymerase Chain Reaction or the like, implementing tubes 2 intended to receive biological samples to be analyzed. Each tube 2 can 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 several treatment step(s), such as a lyophilization operation. Each tube 2 thus pre-filled is sealed using a plug 3 provided with a slot 4 authorizing by elastic deformation the passage of a tip 5 of a pipette.
The method of implementing the device 1 consists of:
The assembly of the lid 10 to the tray 7 is carried out in a simple manner by the approximation of the lid 10 relative to the tray 7. During this approximation, the elastic legs 13a are elastically deformed outwardly until the ribs 13b of the elastic legs 13a penetrate into the housings 14 to be blocked by the abutment surfaces 14a of the tray 7. It should be noted that during this assembly operation, the lid 10 is slidably guided on the tray 7 by the cooperation of the nesting parts of the lid 10 with the nesting parts of the tray 7.
In this assembly position, the lower face 10i of the plate 10p of the lid 10 is positioned above the plugs of the tubes 2 supported by the tray 7 to hold them in position. The lid 10 is bearing on the tray 7, through the rim 10r of its flanged edges 10l, 10t. In this position, the tubes 2 are immobilized or blocked, between the tray 7 and the lid 10 and the chimneys 11 of the lid 10 are positioned above the tubes 2 (
The next step in the preparation of a sample for a Polymerase Chain Reaction or the like consists of introducing the tip 5 of a pipette for injecting a liquid through the chimneys 11 of the lid and the slots 4 of the plugs 3 so as to open out inside the tubes 2 (
Advantageously, the travel of the pipette is limited by the abutments 11c of the chimneys 11 so that the distal end of the tip of the pipette is in the vicinity of or in contact with the wall of the bottom of the tube. The tip of the pipette is ideally positioned in the vicinity of the wall of the tube so as to ensure the operation of deposition of the liquid inside the tube 2. This depositing operation which is an operation delicate to conduct since the liquid to be deposited is generally a drop of small volume typically from 5 to 10 microliters, is thus secured.
After having delivered the liquid in the tubes 2 using the pipette, the operator removes the tip from the pipette, the tubes 2, the plugs 3 and the chimneys 11. The liquid delivered is the sample. A reagent necessary for the amplification can also be added.
It should be noted that the transparency of the tray 7 and the lid 10 gives the operator the possibility of seeing the color code inside the tubes 2.
According to one variant, it should be noted that a film able to be perforated by a tip of a pipette is placed on the lid 10 so as to serve as a fill indicator for filling the tubes 2 with the liquid. This film, not represented in the drawings, can be placed on the upper surface of the chimneys 11 of the lid 10.
According to another variant, before the assembly between the tray 7 and the lid 10, a film able to be perforated by a pipette tip can be interposed between the plugs 3 and the lid 10, so to serve as a fill indicator for filling the tubes 2 with the liquid. This film, not represented in the drawings, can be deposited directly on the upper surface 7s of the tray provided with the plugs 3.
This unitary assembly with the tubes 2 immobilized between the lid 10 and the tray 7 can be introduced into a centrifugation machine of any type known per se to ensure an operation of centrifugation of the contents of the tubes 2. According to one advantage of the device 1 in accordance with the invention, the device 1 can be loaded in a vertical centrifugation machine without the risk of tube leaking. The compact nature of the device 1 facilitates the loading and unloading operations of the device 1.
After carrying out the centrifugation operation, the contents of the tubes 2 are intended to undergo a reaction of amplification of the nucleic acids contained in the tubes using an analysis machine of any type known per se, typically a thermal cycler. According to one variant of implementation, the plugs 3 sealing the tubes 2 are replaced by optical plugs before the operation of analyzing the contents of the tubes.
For this purpose, the lid 10 is separated from the tray 7 and deposited on a laying plane S. This operation is relatively easy to conduct since it suffices to spread the elastic legs 13a to disengage the ribs 13b from the housings 14 and to lift the lid 10 relative to tray 7 according to a translational movement. After bearing the lid 10 on a laying plane S, through its flanged edges 10l, 10t, the tray 7 is placed in a position of superposition above the lid 10 so as to engage the bottom of the tubes protruding from the lower face of the tray in the chimneys 11 of the cover so that they ensure the holding of the tubes. As seen more specifically in
To perform the amplification of the nucleic acids contained in the tubes 2, the unitary assembly consisting of the tube 2 support tray 7 assembled with the lid 10 is loaded in the analysis machine. In the case where the plugs have been replaced by optical plugs, the tray 7 is disengaged from the lid 10 so that the lid can be placed above the tray 7 and fastened together using the removable assembly system 13 as already explained. This unitary assembly makes it easy and safe to load the set of tubes 2 in the analysis machine.
The lid 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. To this end, a bearing element of the analysis machine such as its lid 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 lid of the analysis machine comes into contact with the bearing ribs 7n of the tray 7. The operation of amplification of the nucleic acids contained in the tubes 2 can therefore be conducted by the analysis machine.
It should be noted that during the amplification cycle, the tubes 2 are clamped in the heating block by the bearing element of the analysis machine. At the end of the cycle, it is possible to discharge from the analysis machine, the tubes 2 thanks to the tray 7. This thus allows simultaneously and easily removing the set of tubes 2 having undergone the reaction.
| Number | Date | Country | Kind |
|---|---|---|---|
| FR2006165 | Jun 2020 | FR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/FR2021/051049 | 6/11/2021 | WO |
