BE1029278B1 - Device for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample - Google Patents

Abstract

The present invention relates to a device and a method for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample.

Description

+ BE2021/5267 Device for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample The present invention relates to a device and to a method for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample. More particularly, the present invention relates to a device for detecting the presence of at least one target DNA and/or at least one target RNA in a biological sample, in particular in a liquid biological sample.

In other words, the present invention relates to a device and a method for detecting the presence of at least one target nucleic acid in a sample, in particular in a liquid sample.

As used herein, the terms "nucleic acid" include deoxyribonucleic acid (DNA) and target ribonucleic acid (RNA). The different forms of DNA and RNA are also covered by the term “nucleic acid”.

A device according to the invention finds numerous applications in the fields of health, in particular for detecting, in an individual from whom a sample is simply taken, the presence of at least one target DNA and/or of at least one Target RNA indicating that the individual suffers from one or more pathologies or from one or more given diseases. It may for example be a pathology or a viral disease (viral hepatitis, etc.) or a pathology or a bacterial disease (meningitis, etc.) or even a pathology or a parasitic disease (malaria, etc.).

Devices for detecting target DNA or target RNA are known, some of which allow amplification of target DNA or target RNA to obtain amplicons of the target DNA or of the target RNA (for example by isothermal amplification mediated by loops [LAMP] or by isothermal amplification mediated by reverse transcription loop [RT-LAMP]), in the presence of an agent having redox properties and being capable of binding to amplicons of Target DNA or target RNA: this is referred to as electrochemical measurements using a redox indicator. Such a device known from the state of the art can consist of a receptacle intended to contain a pretreated sample and being associated with electrodes allowing the realization of measurements of the electrochemical type within the pretreated sample in order to detect therein the presence of target DNA or target RNA.

Typically, with current devices for detecting target DNA or target RNA, before placing a sample taken from an individual in such a receptacle and therefore being able to carry out an electrochemical type measurement, the sample must be pretreated, that is to say that it must be placed in solution in the presence of reagents making it possible to carry out an amplification of said target DNA or of said target RNA and in the presence of an agent having oxido-reducing properties and being able to bind to target DNA or target RNA amplicons. In practice, the agent having redox properties and being capable of binding to the target DNA or target RNA amplicons interacts both with the amplicons obtained and with the electrodes to make it possible to carry out an electrochemical measurement and to determine whether or not the sample taken from the individual contains target DNA or target PARN and therefore to determine whether the individual suffers from a given (targeted) pathology or disease. Concretely, in the presence of amplicons obtained if target DNA and/or target RNA is/are present in the sample, a certain quantity of agent exhibiting redox properties binds to the amplicons. of target DNA or target RNA and therefore interacts less / no longer interacts with the electrodes, which results in a modification, in particular a reduction, of the signal measured at the level of the electrodes.

Unfortunately, if such devices are suitable for analyzes carried out in the laboratory, they are not very suitable or even totally unsuitable for detecting target DNA and/or target RNA to be carried out in the field. Indeed, since the pretreatment/preparation of a sample taken from an individual always requires the implementation of numerous reagents, the operator must not only have equipment (reagents, solvents, pipettes, heating device, etc.) ) but also perform many delicate manipulations such as dilutions which must be precise. It is obvious that this is particularly restrictive in the field and that it is even sometimes quite simply impossible to ensure correct preparation of the sample taken, which can lead to erroneous electrochemical measurements (false negatives and/or presence of contaminants, for example). Furthermore, the reagents used are generally sensitive to environmental conditions (high temperatures, humidity, etc.) and/or dangerous to handle, which makes detection of target DNA and/or target RNA all the more restrictive. carried out in the field.

There is therefore currently a real need to provide a device for detecting the presence of target DNA and/or target RNA in a sample, in particular in a liquid sample, which makes it possible to at least partially overcome these problems when detection of one or more target DNAs and/or of one or more target RNAs needs to be carried out, in particular when it needs to be carried out in the field.

To solve these problems, provision is made, according to the invention, for a device for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, said device comprising: - reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP) of said at least one target DNA and/or said at least one target RNA, to obtain amplicons of said at least one target DNA and/or of said at least one target RNA; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA; and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or of said at least a target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification

(LAMP) or carrying out isothermal amplification mediated by reverse transcription loop (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said device intrinsically comprising said reagents allowing amplification of said at least one target DNA and/or said at least one target RNA and intrinsically comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

In other words, there is provided, according to the invention, a device for detecting the presence of at least one target nucleic acid in a sample, in particular in a liquid sample, said device comprising: - reagents allowing the production an amplification of said at least one target nucleic acid to obtain amplicons of said at least one target nucleic acid, for example performing loop-mediated isothermal amplification (LAMP) or performing loop-mediated isothermal amplification reverse transcription (RT-LAMP) of said at least one target nucleic acid, to obtain amplicons of said at least one target nucleic acid; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA; and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the realization of the amplification of said at least one target nucleic acid to obtain amplicons of said at least one target nucleic acid, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound, in particular with said

> BE2021/5267 non-intercalating molecule or with said non-intercalating compound exhibiting oxido-reducing properties, said device intrinsically comprising said reagents allowing amplification of said at least one target nucleic acid and intrinsically comprising said non-intercalating molecule or said compound non-intercalating, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reduction properties.

According to one embodiment, there is provided, according to the invention, a device for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, said device comprising: - reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP) of said at least one target DNA and/or said at least one target RNA, for obtaining amplicons of said at least one target DNA and/or of said at least one target RNA; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA; - an electric heating element arranged to heat said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or said at least one RNA to be carried out target to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing transcription loop-mediated isothermal amplification reverse (RT-LAMP); and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the realization of the amplification of said

© BE2021/5267 at least one target DNA and/or said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example carrying out isothermal amplification mediated by loops (LAMP) or carrying out reverse transcription loop-mediated isothermal amplification (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound, in particular with said non-intercalating molecule or with said non-intercalating compound having redox properties, said device intrinsically comprising said reagents enabling amplification of said at least one target nucleic acid and intrinsically comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound intercalating agent with oxidation-reduction properties.

By definition, the term “intrinsically” means “which is inherent in something, which belongs to it in its own right”. In the context of the present invention, the term "intrinsically" applied to the device according to the invention therefore means that the device comprises, even before its use and therefore before introduction of a sample, inherently and inherently, at least reagents allowing the realization of an amplification of said at least one target DNA and/or of said at least one target RNA and/or at least one non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, or even reagents to lyse said sample.

By the terms “target DNA”, it is understood, within the meaning of the present invention, any type of DNA, for example ribosomal DNA and mitochondrial DNA.

By the terms “target RNA”, it is understood, within the meaning of the present invention, any type of RNA, for example messenger RNA (mRNA), ribosomal RNA (rRNA) and transfer RNA (RNA ).

By the terms “target DNA amplicons”, it is understood, within the meaning of the present invention, the products of an amplification of said target DNA, for example the products of an amplification of said target DNA by the technique of amplification in the polymerase chain (PCR) or by the technique of isothermal loop-mediated amplification (LAMP).

By the terms "target RNA amplicons", it is understood, within the meaning of the present invention, the products of an amplification of said target RNA, in particular the products of an amplification of said target RNA after reverse transcription by implementation of a reverse transcriptase, for example the products of an amplification of said target RNA by the technique of polymerase chain reaction by reverse transcriptase (RT-PCR) or by the technique of isothermal amplification mediated by reverse transcription loop (RT -LAMP).

By the terms "target nucleic acid amplicons", it is understood, within the meaning of the present invention, the products of an amplification of said target nucleic acid, for example the products of an amplification of said target nucleic acid by the technique of polymerase chain reaction (PCR), by the loop-mediated isothermal amplification (LAMP) technique, by the reverse transcriptase polymerase chain reaction (RT-PCR) technique or by the reverse transcription loop-mediated isothermal amplification (RT-LAMP).

By way of example and non-exhaustively, can be implemented as reagents allowing the realization of an amplification of said at least one target DNA and / or said at least one target RNA to obtain amplicons of said at least one Target DNA and/or said at least one target RNA, a reverse transcriptase or retrotranscriptase, a polymerase, a DNA polymerase, primers, dNTPs, and/or buffers.

By the terms "a non-intercalating molecule or a non-intercalating compound", it is understood, within the meaning of the present invention, a molecule or a compound that does not intercalate between DNA or RNA molecules, in particular a molecule or compound that does not fit between two DNA base pairs.

With such a target DNA and/or target RNA detection device according to the invention, the sample taken from an individual does not require pretreatment/preparation, only lysis of the sample being sometimes but not necessarily necessary according to certain embodiments according to the invention, the device according to the invention intrinsically comprising the reagents making it possible to carry out an amplification of said target DNA and/or of said target RNA to obtain amplicons of the target DNA and/or of the target RNA, the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least least one target RNA, optionally an electric heating element and optionally reagents to lyse said sample or a solution to lyse said sample.

The device according to the invention thus allows the operator to dispense with a large quantity of equipment and reagents but also to considerably reduce the manipulations to be carried out in the field to detect a target DNA and/or a target RNA at the start of a sample taken from an individual. The result in particular is that handling errors and contamination are significantly reduced as the electrochemical measurements carried out are all the more reliable and the time required to carry out detection in the field is significantly reduced. In addition, a device for detecting a target DNA and/or a target RNA according to the invention is inexpensive and compact since it does not require large equipment such as those used in the laboratory.

DNA and/or RNA amplification techniques are well known to those skilled in the art. In particular and by way of example, the PCR, RT-PCR, LAMP and RT-LAMP amplification techniques are techniques well known to those skilled in the art who are therefore able to determine, for a given target DNA and/or for a given target RNA, which reagents and which quantities of reagents must be included intrinsically in the device according to the invention (for example: buffers, dNTPs, polymerase, DNA polymerase, primers, reverse transcriptase, etc.).

By way of example, to carry out the detection of the presence of target RNA in a sample, in particular the detection of the presence of target mRNA in a sample, the reagents allowing the realization of an amplification of said target RNA to obtain amplicons of said target RNA comprise a reverse transcriptase or retrotranscriptase and/or a DNA polymerase. Reverse transcriptase or retrotranscriptase is an enzyme used to perform transcription in the reverse of the standard direction (reverse transcription or retrotranscription) to obtain DNA from RNA. reverse transcriptase

? BE2021/5267 therefore makes it possible, starting from RNA, to use amplification techniques such as polymerase chain reaction (PCR) or loop-mediated isothermal amplification (LAMP) conventionally carried out using DNA strands. In other words, using reverse transcriptase, RNA can be transcribed into DNA, making RNA analysis feasible using conventional DNA amplification techniques. When reverse transcriptase or reverse transcriptase is combined with the PCR amplification technique, it is called reverse transcriptase polymerase chain reaction (RT-PCR). When reverse transcriptase or reverse transcriptase is combined with the LAMP amplification technique, it is called reverse transcription loop-mediated isothermal amplification (RT-LAMP).

Note that reverse transcriptase makes it possible to create complementary DNA (cDNA) from mRNA. cDNA is a single strand synthesized from an mRNA, thus representing the coding part of the region of the genome that has been transcribed into this mRNA. cDNA is obtained after a reverse transcription reaction of an mRNA and hence is equivalent to the DNA copy of mRNA. Double-stranded cDNA can result from the copying of a first strand by a DNA polymerase.

Advantageously, according to the invention, said reagents allowing the realization of an amplification of said at least one target DNA and/or of said at least one target RNA are in anhydrous or lyophilized form.

Preferably, according to the invention, said reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA are present on a support comprising positive charges and/or positively charged groups and/or positively charged polymers, for example on a cellulosic support comprising positive charges and/or positively charged groups and/or positively charged polymers.

Advantageously, according to the invention, said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having redox properties, is in anhydrous or lyophilized form.

Preferably, according to the invention, said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having redox properties, is present on a support comprising fillers positive charges and/or positively charged groups and/or positively charged polymers, for example on a cellulosic support comprising positive charges and/or positively charged groups and/or positively charged polymers.

Preferably, the device according to the invention further comprises an electric heating element arranged to heat said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least a target DNA and/or said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example carrying out loop-mediated isothermal amplification (LAMP) or performing reverse transcription loop-mediated isothermal amplification (RT-LAMP).

Advantageously, the device according to the invention further comprises a reagent for lysing said sample or a solution for lysing said sample.

Preferably, according to the invention, said reagent for lysing said sample is in anhydrous or freeze-dried form.

Preferably, the device according to the invention is in the form of a cartridge or a microfluidic device, for example in the form of a cell or a microfluidic chip or a paper-based microfluidic device.

According to the invention, the device may or may not include an electric heating element, in particular may or may not intrinsically comprise an electric heating element. When it does not include an electric heating element, the heating of the device can be ensured, for example, by bringing the device into contact with a heating plate or with any other device making it possible to ensure heating of the sample while the latter is in contact or after the latter has been in contact with said reagents making it possible to carry out the amplification of said at least one target DNA and/or of said at least one target RNA and/or with said non-intercalating molecule or with said non-intercalating compound

H BE2021/5267 intercalating, in particular with said non-intercalating molecule or with said non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

According to the invention, preferably, said electrodes are also arranged to be electrically accessible from the outside of said device. According to the invention, said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having redox properties, capable of binding to the amplicons of the said at least one target DNA and/or to the amplicons said at least one target RNA is selected from the group consisting of Os(bpy)s*; Ru(NHs)e**; FcCH2N*Me3; FcB(OH)2 and mixtures thereof. This list is non-exhaustive and any other non-intercalating molecule or any other non-intercalating compound, in particular any other non-intercalating molecule or any other non-intercalating compound having redox properties, capable of binding to the amplicons of at least one DNA target and/or to the amplicons of at least one target RNA falls within the scope of the present invention. Preferably, the device according to the invention has: - a first part or zone comprising said reagents allowing the performance of an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP); and - a second part or zone, in particular a second part or zone separated from said first part or zone, comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxido- reducing agents, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA. A distinction between these two parts or zones makes it possible to sequence the stages of amplification of said target DNA and/or of said target RNA to obtain amplicons of the target DNA and/or of the target RNA (for example by isothermal amplification mediated by loops [LAMP] or by isothermal amplification mediated by reverse transcription loop [RT-LAMP]) and binding of said non-intercalating molecule or of said non-intercalating compound to the amplicons of said at least one target DNA and/or to the amplicons of said at least a target RNA, which allows the device according to the invention to be optimal, amplicons being first obtained before they come into contact with said non-intercalating molecule or with said non-intercalating compound capable of binding to amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

By way of example, in particular if the device according to the invention is in the form of a chip or a microfluidic cell, the two parts or zones can be distinct chambers fluidically connected (as illustrated in FIGS. 7C and 7D ) or can be separate locations within a single (micro-)channel of a microfluidic chip or cell (as shown in Figure 7F).

According to one embodiment, the device according to the invention for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, comprises: - a first part or zone comprising reagents allowing amplification of said at least one target DNA and/or said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP) of said at least one target DNA and/or said at least one target RNA, to obtain amplicons of said at least one target DNA and/or of said at least one target RNA; - a second part or zone, in particular a second part or zone separated from said first part or zone, comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having redox properties , capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA; and

- electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the realization of the amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing loop-mediated isothermal amplification of reverse transcription (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one DNA target and/or to the amplicons of said at least one target RNA, said device intrinsically comprising said reagents making it possible to carry out an amplification of said at least one target DNA and/or said at least one target RNA and intrinsically comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting oxido-reducing properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

According to another embodiment, the device according to the invention for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, comprises:

- a first part or zone comprising reagents allowing the realization of an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA , for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP) of said at least one target DNA and/or said at least one RNA target, to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, said reagents being in an anhydrous or lyophilized form; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said non-intercalating molecule or said non-intercalating compound being in an anhydrous or lyophilized form and being comprised (contained) in said first part or zone or in a second part or zone, in particular in a second part or zone separated from said first part or zone; and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or of said at least a target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing loop-mediated isothermal amplification reverse transcription (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound, in particular with said non-intercalating molecule or with said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said device intrinsically comprising said reagents making it possible to carry out an amplification of said at least one target DNA and/or said at least one target RNA and intrinsically comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

Advantageously, the device according to the invention comprises a valve situated between said first part or zone and said second part or zone. The presence of a valve makes it possible to control the passage of the sample, lysed or not, from the first part or zone to the second part or zone after a determined residence time of the sample in the first part or zone so that amplifying said target DNA and/or said target RNA to obtain amplicons of target DNA and/or target RNA (e.g. by loop-mediated isothermal amplification [LAMP] or reverse transcription loop-mediated isothermal amplification [RT-LAMP]) is optimized.

Preferably, the device according to the invention further comprises reagents for lysing said sample or a solution for lysing said sample.

In particular, the device according to the invention also intrinsically comprises reagents for lysing said sample or a solution for lysing said sample.

Advantageously, the device according to the invention has an additional part or zone comprising said reagents for lysing said sample or said solution for lysing said sample, said additional part or zone being located upstream of said first part and of said second part in a direction sample flow/displacement.

Depending on the type of sample taken from an individual, it may be useful to lyse the sample. In order to avoid the operator having to carry out a lysis in the field with specific and cumbersome equipment (pipette, lysis solution, etc.), the device according to the invention can advantageously comprise a lysis chamber ( part or additional zone) in which the sample is lysed before reaching the other parts or zones of the device.

Preferably, the device according to the invention comprises a valve situated between said additional part or zone and said first part or zone and/or said second part or zone. The presence of such a valve makes it possible to control the passage from the part or additional zone where the lysis of the sample takes place towards the first and/or the second part after a determined period of residence of the sample in this part or additional area such that sample lysis is optimized.

According to a preferred embodiment of the device according to the invention, said reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of the target DNA and/or of the target RNA (e.g. performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification) and/or said non-intercalating molecule or said non-intercalating compound, in particular said molecule non-intercalating or said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA and/or the reagents for lysing said sample are in non-intercalating form liquid. A non-liquid form can be, for example, a gel, a hydrogel or even a wax, but also can be a non-aqueous dry form such as, for example, a powder.

According to a preferred embodiment of the device according to the invention, said reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of the target DNA and/or of the target RNA (e.g. performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification) and/or said non-intercalating molecule or said non-intercalating compound, in particular said molecule non-intercalating or said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA and/or the reagents for lysing said sample are in a form anhydrous or freeze-dried.

In particular, according to an advantageous embodiment of the device according to the invention, said reagents making it possible to carry out an amplification and said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting redox, are in an anhydrous or freeze-dried form.

In particular, according to another advantageous embodiment of the device according to the invention, said reagents making it possible to carry out an amplification and said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reduction properties, and said reagents for lysing said sample are in an anhydrous or freeze-dried form.

Unlike liquid solutions comprising the reagents allowing amplification of said target DNA and/or said target RNA and/or said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reduction properties, and/or the reagents for lysing said sample, an anhydrous or freeze-dried form of the latter makes it possible to have a device which can be kept for several months or even several years without degradation of the reagents allowing amplification to be carried out and/or said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting oxido-reducing properties, and/or reagents for lysing said sample, which retain their properties during the time, which contributes to the reproducibility of the results.

Current devices systematically make use of a liquid form of the reagents allowing amplification of said target DNA and/or said target RNA and/or a liquid form of said non-intercalating molecule or said non-intercalating compound, in particular at a liquid form of reagents allowing amplification of said target DNA and/or said target RNA and/or a liquid form of said non-intercalating molecule or said non-intercalating compound introduced into the device after introduction into the latter of a sample (the reagents making it possible to carry out an amplification of said target DNA and/or of said target RNA and the non-intercalating molecule or the non-intercalating compound are therefore not intrinsically present in the device). With current devices, this liquid form of the reagents allowing the realization of an amplification of said target DNA and/or of said target RNA and/or this liquid form of the non-intercalating molecule or of the non-intercalating compound is systematically used and recommended to ensure the availability of these compounds. In particular, the reagents must be able to interact with the target DNA and/or the target VRNA and the non-intercalating molecule or the non-intercalating compound must be able to interact both with the target DNA and/or the target RNA and with the electrodes (especially with the working electrode) so that correct and reliable electrochemical measurements can be made.

Unlike current devices, said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to amplicons of said at least one target RNA is present in a device according to the invention in a non-liquid form, in particular in an anhydrous or lyophilized form, and it has been shown, in the context of the present invention and at the contrary to what is recommended and encountered with the devices available today, that this non-liquid form, in particular this anhydrous or freeze-dried form, of said non-intercalating molecule or of said non-intercalating compound, in particular of said non-intercalating molecule or of said non-intercalating compound having redox properties, makes it possible to carry out precise and reliable measurements to determine whether a bond is t / has been established between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound, in particular between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound exhibiting oxidation-reduction properties, in order to deduce that said at least one target DNA and/or said at least one target RNA is present or not in said sample and to ensure detection of said at least one at least one target DNA and/or said at least one target RNA with a device according to the invention.

In other words, against all expectation, it has been demonstrated in the context of the present invention that a non-liquid form of said non-intercalating molecule or of said non-intercalating compound, in particular an anhydrous or freeze-dried form of said non-intercalating molecule or said non-intercalating compound, intrinsically present in a device according to the invention, indeed makes it possible to carry out precise and reliable measurements to determine whether a bond is/has been established between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound, in particular between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound exhibiting oxidation-reduction properties , in order to deduce, via an electrochemical measurement, that said at least one target DNA and/or said at least one target RNA is present or not in said sample and to ensure a detection of said at least one target DNA and/or of said at least one target RNA with a device according to the invention.

According to an embodiment according to the invention, the device for detecting the presence of at least one target DNA and/or of at least one target RNA in a sample, in particular in a liquid sample, comprises: - reagents allowing carrying out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example carrying out an isothermal amplification mediated by loops (LAMP) or carrying out isothermal amplification mediated by reverse transcription loop (RT-LAMP) of said at least one target DNA and/or of said at least one target RNA, in order to obtain amplicons of said at least one target DNA and/or said at least one target RNA, said reagents being in an anhydrous or lyophilized form; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said non-intercalating molecule or said non-intercalating compound being in an anhydrous or lyophilized form; and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or of said at least a target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing loop-mediated isothermal amplification reverse transcription (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound, in particular with said non-intercalating molecule or with said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said device intrinsically comprising said reagents making it possible to carry out an amplification of said at least one target DNA and/or said at least one target RNA and intrinsically comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

According to another embodiment, the device according to the invention for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, comprises: - a first part or zone comprising reagents allowing amplification of said at least one target DNA and/or said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP) of said at least one target DNA and/or said at least one target RNA, for obtaining amplicons of said at least one target DNA and/or of said at least one target RNA, said reagents being in an anhydrous or lyophilized form; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said non-intercalating molecule or said non-intercalating compound being in an anhydrous or lyophilized form and being comprised (contained) in said first part or zone or in a second part or zone; and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or of said at least a target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing loop-mediated isothermal amplification reverse transcription (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound, in particular with said non-intercalating molecule or with said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, said device intrinsically comprising said reagents making it possible to carry out an amplification of said at least one target DNA and/or said at least one target RNA and intrinsically comprising said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

Preferably according to the invention, said reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of the target DNA and/or of the target RNA and /or said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least a target RNA, and/or said reagents for lysing said sample are in an anhydrous or lyophilized form in the device according to the invention.

Preferably according to the invention, said reagents making it possible to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of the target DNA and/or of the target RNA and /or said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least a target RNA, and/or said reagents for lysing said sample are in the form of a lyophilisate in the device according to the invention.

According to an advantageous embodiment of the device according to the invention, said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reducing properties, capable of binding to the amplicons of said at least a target DNA and/or to the amplicons of said at least one target RNA and/or said reagents making it possible to carry out an amplification of said target DNA and/or of said target RNA to obtain amplicons of the target DNA and/or of the Target RNAs, for example performing loop-mediated isothermal amplification (LAMP) or performing reverse-transcription loop-mediated isothermal amplification (RT-LAMP), are present on a support having positive charges and/or or positively charged groups and/or positively charged polymers, for example on a cellulosic support comprising positive charges and/or positively charged groups and/or positively charged polymers.

For example, positively charged groups are associated with a support, in particular with a cellulosic support, by covalent or other grafting.

Alternatively or additionally, positively charged polymers are associated with a support, in particular with a cellulosic support, by adsorption.

Alternatively or additionally, predetermined chemical species are polymerized on the surface of a support, in particular on the surface of a cellulosic support, so that positively charged polymers are associated with this support.

In the context of the present invention, it has been put forward, in particular for said non-intercalating molecule or said non-intercalating compound, in particular for said non-intercalating molecule or said non-intercalating compound having oxido-reducing properties, capable of bind to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, that such a positively charged support makes it possible to guarantee adequate availability thereof, in the sense that it does not remain attached to the support in order ensure adequate electrochemical measurement.

Advantageously, according to the invention, said sample is chosen from a blood sample, a saliva sample, a urine sample.

The sample can be in liquid or semi-liquid form but also in solid or semi-solid form.

Preferably, according to the invention, said electrical heating element comprises an electrical resistor, which is powered by a source of electrical energy.

Preferably, according to the invention, said electrodes are three in number.

Preferably, the device according to the invention is in the form of a cartridge or a microfluidic device, for example in the form of a cell or a microfluidic chip or a paper-based microfluidic device.

Advantageously, the device according to the invention is in the form of a cartridge through which a solution or a microfluidic device can flow, for example in the form of a cell or a microfluidic chip. The device according to the invention can also take the form of a paper-based microfluidic device (“paper-based microfluidics”) consisting for example of a series of hydrophilic cellulose or nitrocellulose fibers which guide a liquid from one entry to a desired area by imbibition and/or capillary action. Optionally, the paper-based microfluidic device according to the invention is in the form of an origami ("paper-origami based"), that is to say in the form of a microfluidic paper of which parts are intended to be folded one on the other or on each other to be brought into contact. Within the meaning of the present invention, the device may also preferentially take the form of a “paper-based point-of-care NAATs (Nucleic acid amplification-based tests)”.

The present invention also relates to an assembly comprising: - a device according to the invention; and - an apparatus or apparatus that can be electrically connected to the electrodes to perform electrochemical measurements, preferably electrochemical voltammetric measurements, more preferably cyclic electrochemical voltammetric measurements or square waves.

Typically, to carry out electrochemical measurements, preferably electrochemical voltammetric measurements, more preferably cyclic or square wave electrochemical voltammetric measurements, the apparatus comprises a potentiostat for applying a potential difference between the electrodes and/or a processing unit of signals emitted by the potentiostat or received by the potentiostat.

The present invention also relates to a method for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, said method comprising the following steps: - a step of introducing said sample into a device according to the invention or into a device of an assembly according to the invention; - a first step of bringing said sample into contact, within said device, with reagents intrinsically present in said device to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least a target DNA and/or said at least one target RNA; - a second step of bringing into contact, within said device, during or after said first bringing into contact, said sample with a non-intercalating molecule or with a non-intercalating compound intrinsically present in said device, in particular with a molecule non-intercalating or with a non-intercalating compound intrinsically present in said device having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA; - a third step of bringing into contact, within said device, during or after said first bringing into contact and/or during or after said second bringing into contact, said sample with electrodes electrically connected to a device or to an apparatus for carrying out at least one electrochemical measurement, to determine whether a bond is/has been established between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound, in particular between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound having redox properties, in order to deduce that said at least one target DNA and/or said at least one target RNA is present or not in said sample and to ensure detection of said at least one target DNA and/or of said at least one target RNA.

Preferably, the method according to the invention comprises heating said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA.

Advantageously, according to the invention, said first step of bringing into contact and/or said second step of bringing into contact consists of dissolving by said sample said reagents intrinsically present in said device to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or amplicons of said at least one target RNA, and/or in solution by said sample of said non-intercalating molecule or of said compound non-intercalating intrinsically present in said device, in particular by dissolving said non-intercalating molecule or said non-intercalating compound intrinsically present in said device and having oxido-reducing properties, to allow the establishment of a bond between said amplicons of said at least one target DNA and/or said amplicons of said at least one target RNA and said non-intercalating molecule e or said non-intercalating compound, in particular to allow the establishment of a bond between said amplicons of said at least one target DNA and/or said amplicons of said at least one target RNA and said non-intercalating molecule or said non-intercalating compound exhibiting redox properties.

Preferably, the method according to the invention comprises a step of lysis of said sample, preferably a step of lysis of said sample carried out upstream of said first and/or of said second contacting step.

The present invention also relates to the use of a device according to the invention or of an assembly according to the invention for detecting the presence of at least one target DNA and/or at least one target RNA in a sample. .

Other characteristics, details and advantages of the invention will emerge from the examples given below, on a non-limiting basis and with reference to the appended figures. Generally, in the figures, similar elements bear the same reference.

Figure 1 illustrates a first example of an embodiment of a device according to the invention.

Figure 2 illustrates a second example of an embodiment of a device according to the invention.

Figure 3 illustrates a third example of an embodiment of a device according to the invention.

Figure 4 illustrates a fourth example of an embodiment of a device according to the invention.

Figures 5A, 5B and 5C are exploded views of exemplary embodiments of devices according to the invention and illustrate the components of different embodiments of a device according to the invention.

Figures 6A, 6B, 6C, 6D and 6E are exploded views of exemplary embodiments of devices according to the invention and also illustrate the components of different embodiments of a device according to the invention.

FIGS. 7A, 7B, 7C, 7D, 7E and 7F further illustrate examples of embodiments of devices according to the invention.

Examples FIG. 1 illustrates a first embodiment of a device according to the invention which, as illustrated, comprises a first part or zone 1, a second part or zone 2, electrodes 3 and an electric heating element 4. Optionally, as illustrated by the dotted lines, this embodiment of a device according to the invention can comprise an additional part or zone 5 comprising reagents for lysing the sample or a solution for lysing a sample taken from an individual. FIG. 1 illustrates an example of a device according to the invention which could be in the form of a cartridge or of a microfluidic device, for example in the form of a microfluidic device based on paper (“paper- based microfluidics” or “paper-origami based” or “paper-based point-of-care NAATS”).

The device according to the invention intrinsically comprises in a first part or zone 1, reagents allowing the realization of an amplification of said target DNA and/or of said target RNA to obtain amplicons of the target DNA and/or of the RNA target potentially contained in the sample (e.g. performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification). The device also intrinsically comprises in a second part or zone 2 a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having oxidation-reducing properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA. With such a device according to the invention, the sample taken from an individual is preferably lysed, that is to say dissolved in a lysis solution, before being introduced into the device as indicated by the arrow. When, for example under the effect of gravity, the lysed sample reaches the first part or zone 1, it comes into contact with the reagents making it possible to carry out an amplification of said target DNA and/or of said target RNA to obtain amplicons of target DNA and/or target RNA (e.g. performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification), in the first part or zone 1 under the effect of heating to a predetermined temperature by the electric heating element 4. Amplicons of the target DNA and/or of the target RNA are thus obtained provided that this target DNA and/or that this target RNA is present in the sample taken at the departure of the individual. The amplicons obtained then gain, for example under the effect of gravity, the second part or zone 2 comprising a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties , capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

According to the illustrated embodiment, the electrodes 3 (working electrode, counter-electrode or auxiliary electrode and reference electrode) are in contact with the sample after the latter has been in contact with said reagents allowing the realization of the amplifying said target DNA and/or said target RNA to obtain target DNA and/or target RNA amplicons (for example performing isothermal loop-mediated amplification or performing isothermal mediated amplification by reverse transcription loop), and with said non-intercalating molecule or with said non-intercalating compound, in particular with said non-intercalating molecule or with said non-intercalating compound exhibiting oxido-reducing properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA. The electrodes 3 are also electrically accessible from the outside of the device according to the invention so that they can be electrically connected to an apparatus (not shown) making it possible to carry out electrochemical measurements (preferably electrochemical voltammetric measurements, more preferably cyclic or square wave voltammetric electrochemical measurements) using a redox indicator (non-intercalating molecule or non-intercalating compound), the principle and implementation of such an electrochemical measurement being well known to those skilled in the art.

When the embodiment illustrated in FIG. 1 also comprises an additional part or zone 5 comprising a solution or reagents for lysing a sample taken from an individual, the sample can be introduced directly into the device at the level of the additional part or zone 5 in which the sample is lysed before reaching, for example under the effect of gravity, the first part or zone 1 then the second part or zone2.

Figure 2 illustrates a second embodiment of a device according to the invention. FIG. 2 illustrates an example of a device according to the invention which could be in the form of a cartridge or of a microfluidic device, for example in the form of a paper-based microfluidic device (“paper- based microfluidics” or “paper-origami based” or “paper-based point-of-care NAATs”). The same elements as those illustrated in FIG. 1 are repeated. However, according to the embodiment as illustrated in FIG. 2, the first part or zone 1 and the second part or zone 2 are located at the same level, that is to say in the same zone or the same compartment. For example, the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least minus one

Target RNA, located in the second part or zone 2, can coat the side walls of a zone or a compartment of the device, thus surrounding the first part or zone 1 comprising the reagents allowing the amplification to be carried out of said target DNA and/or said target RNA to obtain amplicons of target DNA and/or target RNA potentially contained in the sample (for example performing loop-mediated isothermal amplification or performing isothermal amplification mediated by reverse transcription loop). It is not excluded, according to the invention, that the reagents allowing the amplification of said target DNA and/or said target RNA and that said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA, are present as a mixture in the same zone or in the same compartment, which would then comprise said first and said second part or zone.

Figure 3 illustrates a third embodiment of a device according to the invention. FIG. 3 illustrates an example of a device according to the invention which could be in the form of a cartridge or of a microfluidic device, for example in the form of a microfluidic device based on paper (“paper- based microfluidics” or “paper-origami based” or “paper-based point-of-care NAATS”). Unlike the embodiment illustrated in FIG. 1, that illustrated in FIG. 3 comprises an additional part or zone 7 located upstream and in contact with the electrodes 3 in a flow direction of the sample.

The embodiment illustrated in FIG. 4 is identical to that of FIG. 3 except for the fact that the heating provided by the electric heating element 4 takes place at the level of the additional part or zone 7 located upstream and in contact with the electrodes 3 in a flow direction of the sample rather than at the level of the first part or zone 1. FIG. 4 illustrates an example of a device according to the invention which could be in the form of a cartridge or a microfluidic device, for example in the form of a paper-based microfluidic device (“paper-based microfluidics” or “paper-origami based” or “paper-based point-of-care NAATS”).

Figures 5A, 5B and 5C illustrate the components of different embodiments of a device according to the invention.

More particularly, FIGS. 5A, 5B and 5C illustrate the components of a device A according to the invention which is in the form of a cylinder constituting for example a cartridge.

In FIG. 5A, the device A according to the invention comprises a first part consisting for example of a 1+2 permeable disc, for example of a permeable disc made of cellulosic material, comprising both the reagents allowing the production of the amplifying said target DNA and/or said target RNA to obtain amplicons of the target DNA and/or target RNA (for example performing loop-mediated isothermal amplification or performing isothermal amplification mediated by reverse transcription loop), and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having oxidoreductive properties, capable of binding to the amplicons of the said at least one target DNA and / or to the amplicons of said at least one target RNA.

An electrical heating element (not shown) provides heating of the 1+2 permeable disc to a predetermined temperature to ensure the amplification of said target DNA and/or said target RNA to obtain amplicons of the target DNA and/or the RNA target (e.g. loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification). According to the embodiment of FIG. 5A, three electrodes 3 are in contact with the sample while the latter is in contact with said reagents enabling the amplification of said target DNA and/or said target RNA to be carried out. to obtain amplicons of target DNA and/or target DNA (e.g. performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification), and with said molecule not intercalating compound or with said non-intercalating compound, in particular with said non-intercalating molecule or with said non-intercalating compound having redox properties, capable of binding to amplicons of said at least one target DNA and/or to amplicons of said at least one RNA target.

In addition, the electrodes 3 are also electrically accessible from outside the device according to the invention so that they can be electrically connected to an apparatus (not shown) making it possible to carry out electrochemical measurements (preferably electrochemical voltammetric measurements , more preferably cyclic or square-wave voltammetric electrochemical measurements) using a redox indicator (non-intercalating molecule or non-intercalating compound), the principle and implementation of such an electrochemical measurement being well known to those skilled in the art. As illustrated in FIG. 5A, the electrodes 3 of the device A according to the invention can be electrically connected to an apparatus (not illustrated) via a base B or a base provided with connectors. For example, the device A according to the invention can be screwed or clipped onto the base or the base B to ensure adequate contact between the connectors and the electrodes 3.

FIG. 5B is identical to FIG. 5A but the device A illustrated also comprises an additional part or zone 5 comprising reagents or a solution for lysing a sample taken from an individual. In this way, the sample can be introduced directly into the device at the level of the additional part or zone 5 in which the sample is lysed before reaching, for example under the effect of gravity, the part consisting for example of a 1+2 permeable disk comprising both the reagents allowing the amplification of said target DNA and/or said target RNA to obtain amplicons of target DNA and/or target RNA (for example the production of loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification), and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having oxidative properties -reductive, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

FIG. 5C is identical to FIG. 5B but the device A illustrated additionally comprises a valve 6, for example a rotary slide valve, making it possible to control the passage of the sample from the part or additional zone 5 where it is lysed towards the part consisting for example of a 1+2 permeable disk comprising both the reagents allowing the amplification of said target DNA and/or said target RNA to obtain amplicons of the target DNA and/or of the target RNA (e.g. performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification), and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound exhibiting redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

The devices illustrated in FIGS. 5A, 5B and 5C intrinsically comprise the reagents making it possible to carry out the amplification of said target DNA and/or of said target RNA and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the compound non-intercalating having oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA and, optionally reagents or a solution for lysing a sample taken from a individual.

Figures 6A, GB, 6C, 6D and 6E also illustrate the components of different embodiments of a device according to the invention. More particularly, FIGS. 6A, 6B, 6C, 6D and GE illustrate the components of a device A according to the invention which is in the form of a cylinder constituting for example a cartridge. In figure GA, the device A according to the invention comprises: - a first part 1 consisting for example of a permeable disc, for example of a permeable disc made of cellulosic material, comprising the reagents allowing the realization of the amplification of said target DNA and/or said target RNA to obtain amplicons of target DNA and/or target RNA (e.g. performing loop-mediated isothermal amplification or performing loop-mediated isothermal amplification reverse transcription); and - a second part 2 consisting for example of a permeable disk, for example a permeable disk of cellulosic material, comprising the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

An electric heating element (not shown) heats at least the permeable disc 1 to a predetermined temperature to ensure the amplification of said target DNA and/or said target RNA to obtain amplicons of the target DNA and/or target RNA (e.g. to provide loop-mediated isothermal amplification or to provide reverse transcription loop-mediated isothermal amplification). According to the embodiment of FIG. GA, three electrodes 3 are in contact with the sample after the latter has been in contact with said reagents allowing amplification of said target DNA and/or said RNA to be carried out. target to obtain amplicons of the target DNA and/or target RNA (e.g., performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification), and with the non-intercalating molecule or with the non-intercalating compound, in particular with the non-intercalating molecule or with the non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least least one target RNA. In addition, the electrodes 3 are also electrically accessible from outside the device according to the invention so that they can be electrically connected to an apparatus (not shown) making it possible to carry out electrochemical measurements (preferably electrochemical voltammetric measurements , more preferably cyclic or square wave voltammetric electrochemical measurements) using a redox indicator (non-intercalating molecule or non-intercalating compound), the principle and implementation of such an electrochemical measurement being well known to those skilled in the art. As illustrated in figure GA, the electrodes 3 of the device A according to the invention can be electrically connected to an apparatus (not illustrated) via a base B or a base provided with connectors. For example, the device A according to the invention can be screwed or clipped onto the base or the base B to ensure adequate contact between the connectors and the electrodes 3.

FIG. 6B is identical to FIG. 6A but a valve 6, for example a rotary slide valve, separates the first part 1 from the second part 2 and makes it possible to control the passage of the sample from the first part 1 comprising the reagents allowing the realization of the amplification of said

target DNA and/or said target RNA to obtain amplicons of target DNA and/or target RNA (e.g. performing loop-mediated isothermal amplification or performing loop-mediated isothermal amplification reverse transcription), to the second part 2 comprising the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having oxidoreductive properties, capable of binding to the amplicons of the said at least one DNA target and/or to the amplicons of said at least one target RNA.

FIG. 6C is identical to FIG. 6A but the device A illustrated also comprises an additional part or zone 5 comprising reagents or a solution for lysing a sample taken from an individual. In this way, the sample can be introduced directly into the device at the level of the additional part or zone 5 in which the sample is lysed before reaching, for example under the effect of gravity, the first part 1 constituted by example of a permeable disk comprising the reagents allowing the amplification of said target DNA and/or said target RNA to obtain amplicons of the target DNA and/or of the target RNA (for example the production of a loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification), then to the second part 2 comprising the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA. FIG. 6D is identical to FIG. 6C but the device A illustrated additionally comprises a valve 6′, for example a rotary slide valve, making it possible to control the passage of the sample from the part or additional zone 5 where it is lysed. to the first part 1 consisting for example of a permeable disk comprising the reagents allowing the realization of the amplification of said target DNA and/or of said target RNA to obtain amplicons of target DNA and/or of target RNA (for example the realization of an isothermal amplification mediated by loops or the production of an isothermal amplification mediated by a reverse transcription loop) then to the second part 2 comprising the non-intercalating molecule or the non-intercalating compound,

in particular the non-intercalating molecule or the non-intercalating compound exhibiting oxidation-reduction properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

FIG. 6E is identical to FIG. 6D but the device A illustrated additionally comprises a valve 6, for example a rotary slide valve, making it possible to control the passage of the sample from the first part 1 consisting for example of a disc permeable cell comprising the reagents allowing the realization of the amplification of said target DNA and/or of said target RNA to obtain amplicons of the target DNA and/or of the target RNA (for example the realization of an isothermal amplification mediated by loops or carrying out isothermal amplification mediated by reverse transcription loop), to the second part 2 comprising the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties , capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

The devices illustrated in FIGS. GA, 6B, 6C, 6D and 6E intrinsically comprise the reagents allowing the amplification of said target DNA and/or said target RNA and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule, to be carried out. intercalating compound or the non-intercalating compound exhibiting redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA and, optionally reagents or a solution for lysing a sample taken from the departure of an individual.

FIGS. 7A, 7B, 7C, 7D, 7E and 7F illustrate examples of embodiments of devices according to the invention in the form of microfluidic devices (cells or microfluidic chips) having an inlet E via which a sample can be introduced. Elements identical to those illustrated in the preceding figures are repeated.

In particular, FIG. 7E illustrates a device according to the invention as illustrated in FIGS. 7A to 7D with the difference that the electrodes are in direct contact with an area comprising both the reagents allowing the amplification of said target DNA and/or said target RNA to obtain amplicons of target DNA and/or target RNA (e.g. performing loop-mediated isothermal amplification or performing transcription loop-mediated isothermal amplification inverse), and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA.

According to the embodiment of a device according to the invention illustrated in FIG. 7F, the (micro-)channel C of a microfluidic cell or chip constitutes a first zone or part of the device, this first zone or part comprising a first sub-zone (delimited by the dotted lines) where the reagents 1 are found, allowing the amplification of said target DNA and/or said target RNA to obtain amplicons of the target DNA and/or of the target RNA (for example the performing loop-mediated isothermal amplification or performing reverse transcription loop-mediated isothermal amplification) and a second subzone (delimited by the other dotted lines) where the non-intercalating molecule or the non-intercalating compound is located , in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one RNA target, each of these two sub-zones being located in said first zone or part of the device consisting of said (micro-)channel of a microfluidic device and therefore being in fluid communication. The devices illustrated in FIGS. 7A, 7B, 7C, 7D, 7E and 7F intrinsically comprise the reagents enabling the amplification of said target DNA and/or said target RNA and the non-intercalating molecule or the non-intercalating compound, in particular the non-intercalating molecule or the non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA and, optionally reagents or a solution for lysing a sample taken from an individual. The present invention has been described in relation to specific embodiments, which are purely illustrative and should not be construed as limiting. In general, it will appear obvious to those skilled in the art that the present invention is not limited to the examples illustrated and/or described above.

The use of the verbs "understand", "include", "compose", or any other variant, as well as their conjugations, can in no way exclude the presence of elements other than those mentioned. The use of the indefinite article "un", "une", or of the definite article "le", "la" or "l'", to introduce an element does not exclude the presence of a plurality of these elements.

Claims (15)

Claims 1. Device for detecting the presence of at least one target DNA and / or at least one target RNA in a sample, in particular in a liquid sample, said device comprising: - reagents allowing the realization of an amplification of said at least one target DNA and/or said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or carrying out reverse transcription loop-mediated isothermal amplification (RT-LAMP) of said at least one target DNA and/or said at least one target RNA, to obtain amplicons of said at least one target DNA and/or said at least one least one target RNA; - a non-intercalating molecule or a non-intercalating compound, in particular a non-intercalating molecule or a non-intercalating compound having redox properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA; and - electrodes arranged to be in contact with said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the amplification of said at least one target DNA and/or of said at least a target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example performing loop-mediated isothermal amplification (LAMP) or performing loop-mediated isothermal amplification reverse transcription (RT-LAMP), and with said non-intercalating molecule or with said non-intercalating compound capable of binding to amplicons of said at least one target DNA and/or to amplicons of said at least one target RNA, said device intrinsically comprising said reagents allowing amplification of said at least one target DNA and/or said at least one target RNA and intrinsically comprising said non-intercalating molecule or said compound non-intercalating molecule, in particular said non-intercalating molecule or said non-intercalating compound having oxidation-reducing properties, capable of binding to the amplicons of said at least one target DNA and/or to the amplicons of said at least one target RNA. 2. Device according to claim 1, characterized in that said reagents allowing the realization of an amplification of said at least one target DNA and/or of said at least one target RNA are in anhydrous or lyophilized form. 3. Device according to claim 1 or 2, characterized in that said reagents allowing the realization of an amplification of said at least one target DNA and / or of said at least one target RNA are present on a support comprising positive charges and / or positively charged groups and/or positively charged polymers, for example on a cellulosic support comprising positive charges and/or positively charged groups and/or positively charged polymers. 4. Device according to any one of the preceding claims, characterized in that said non-intercalating molecule or said non-intercalating compound, in particular said non-intercalating molecule or said non-intercalating compound having oxido-reducing properties, is in anhydrous or lyophilized form. . 5. Device according to any one of the preceding claims, characterized in that the said non-intercalating molecule or the said non-intercalating compound, in particular the said non-intercalating molecule or the said non-intercalating compound having oxido-reducing properties, is present on a support comprising positive charges and/or positively charged groups and/or positively charged polymers, for example on a cellulosic support comprising positive charges and/or positively charged groups and/or positively charged polymers. 6. Device according to any one of the preceding claims, characterized in that it further comprises an electric heating element arranged to heat said sample while the latter is in contact or after the latter has been in contact with said reagents allowing carrying out the amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA, for example carrying out isothermal amplification loop-mediated amplification (LAMP) or performing reverse transcription loop-mediated isothermal amplification (RT-LAMP). 7. Device according to any one of the preceding claims, characterized in that it further comprises a reagent for lysing said sample or a solution for lysing said sample. 8. Device according to claim 7, characterized in that said reagent for lysing said sample is in anhydrous or freeze-dried form. 9. Device according to any one of the preceding claims, characterized in that it is in the form of a cartridge or a microfluidic device, for example in the form of a cell or a microfluidic chip or a paper-based microfluidic device. 10. Assembly comprising: - a device according to any one of claims 1 to 9; and - an apparatus or apparatus that can be electrically connected to the electrodes to perform electrochemical measurements, preferably electrochemical voltammetric measurements, more preferably cyclic electrochemical voltammetric measurements or square waves. 11. Method for detecting the presence of at least one target DNA and/or at least one target RNA in a sample, in particular in a liquid sample, said method comprising the following steps: - a step of introducing said sample into a device according to any one of claims 1 to 9 or into a device of an assembly according to claim 10; - a first contacting step, within said device, said sample with reagents intrinsically present in said device to carry out an amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of said at least one target RNA; - a second contacting step, within said device, during or after said first bringing into contact, said sample with a non-intercalating molecule or with a non-intercalating compound intrinsically present in said device, in particular with a non-intercalating molecule or with a non-intercalating compound intrinsically present in said device having redox properties, capable of binding to amplicons of said at least one target DNA and/or to amplicons of said at least one target RNA; - a third step of bringing into contact, within said device, during or after said first bringing into contact and/or during or after said second bringing into contact, said sample with electrodes electrically connected to a device or to an apparatus for carrying out at least one electrochemical measurement, to determine whether a bond is/has been established between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound, in particular between said at least one target DNA and/or said at least one target RNA and said non-intercalating molecule or said non-intercalating compound having redox properties, in order to deduce that said at least one target DNA and/or said at least one target RNA is present or not in said sample and to ensure detection of said at least one target DNA and/or of said at least one target RNA. 12. Method according to claim 11, characterized in that it comprises heating of said sample while the latter is in contact or after the latter has been in contact with said reagents allowing the realization of the amplification of said at least one DNA target and/or said at least one target RNA to obtain amplicons of said at least one target DNA and/or said at least one target RNA. 13. Method according to claim 11 or 12, characterized in that said first contacting step and/or said second contacting step consists of dissolving by said sample said reagents intrinsically present in said device to carry out a amplification of said at least one target DNA and/or of said at least one target RNA to obtain amplicons of said at least one target DNA and/or of amplicons of said at least one target RNA, and/or in solution by said sample of said non-intercalating molecule or said non-intercalating compound intrinsically present in said device, in particular by dissolving said non-intercalating molecule or said non-intercalating compound intrinsically present in said device and having redox properties , to allow the establishment of a link between said amplicons of said at least one target DNA and/or said amplicons of said at least one target RNA and said non-intercalating molecule or said non-intercalating compound, in particular to allow the establishment of a bond between said amplicons of said at least one target DNA and/or said amplicons of said at least one target RNA and said non-intercalating molecule or said non-intercalating compound intercalating agent with oxidation-reduction properties. 14. Method according to any one of claims 11 to 13, characterized in that it comprises a step of lysis of said sample, preferably a step of lysis of said sample carried out upstream of said first and/or of said second step of put in contact. 15. Use of a device according to any one of claims 1 to 9 or of an assembly according to claim 10 for detecting the presence of at least one target DNA and/or at least one target RNA in a sample .