Field reader of biological samples processed by loop-mediated isothermal DNA amplification

Abstract

Device (100) for carrying out and reading the loop-mediated isothermal amplification reaction on a sample to be analyzed received in a reaction tube (T), Figure for the abstract: 2

Description

Title of the invention: Field reader of biological samples processed by loop-mediated isothermal DNA amplification Technical field

[0001] The present invention applies to the production of a field reader, in English point of care test, of biological samples treated by loop-mediated isothermal amplification of DNA (LAMP). State of the prior art

[0002] In the broad sense, a field reader, designated by the acronym POCT, for point of care tests, is a medical laboratory diagnostic test intended to be carried out in the immediate vicinity of the patient, in a doctor's office, in pharmacies, polyclinics or medical centers, in the emergency rooms of certain hospital establishments, or even in professional hospital or non-hospital laboratories, on the condition that the result can be obtained within a short time. These tests are designed to be carried out by personnel not necessarily trained in laboratory medicine (nurse, medical assistant), or even by the patient himself or his relatives.

[0003] Modern medicine has been using polymerase chain reaction techniques for many years to amplify DNA, for a particular target deoxyribonucleic acid, contained in a sample and to detect its presence.

[0004] Polymerase chain reaction, known as ACP in French, also known as polymerase chain reaction, or PCR for English Polymerase Chain Reaction, or nucleic acid amplification test TAN, in French-speaking Canada, is a molecular biology method of in vitro gene amplification.

[0005] This method typically requires a series of high and low temperature cycles to be performed, each cycle allowing for a new amplification step. The series of cycles typically lasts 3 to 5 hours so that sufficient copies are available for analysis. Typically, a fluorophore, i.e., a chemical substance capable of emitting fluorescence light after excitation by light of a lower wavelength, is inserted into each copy so that the quantity and spectrum of light re-emitted by the fluorophore can be detected by illumination from a light source.

[0006] Complex and expensive equipment is required to carry out this operation. Since the amplification time is significant, this equipment is constructed so that it can process a large number of samples at the same time.

[0007] Many methods, such as that described in patent US6605813, are used to optimize the design of this equipment.

[0008] An invention by the company EIKEN in 2000 described an alternative method for amplifying DNA or RNA (ribonucleic acid) which has the following advantages over the method:

[0009] • allow amplification at constant temperature, • benefit from a shorter reaction time of around 20 to 45 minutes, • be potentially usable with simplified equipment compared to the complex and expensive equipment traditionally used.

[0010] The references in this publication are as follows: Tsugunori & Okayama, Hiroto & Masubuchi, Harumi & Yonekawa, Toshihiro & Watanabe, Keiko & Amino, Nobuyuki & Hase, Tetsu. (2000). Loop-mediated isothermal amplification of DNA. Nucleic acids research. 28.E63. 10.1093 / nar / 28.12.e63.)

[0011] A principle for producing such equipment is described by the publication of patent application US2020063197.

[0012] With reference to Figure 1, such equipment S comprises a heating stage SI provided with a heat source and light sources, arranged to receive a plurality of samples in wells S2, a colorimetric camera S3 arranged opposite the heating stage SI and a computer S4 arranged to receive computer program products comprising instructions for analyzing the data generated by the camera S3 when they are executed by the computer S4.

[0013] The S equipment, even if it is capable of being miniaturized, has the disadvantage of remaining bulky and unlikely to be used in the field. It is also expensive given the presence of a colorimetric camera to analyze the results.

[0014] Due to the operations it involves for diagnosis (presence or absence of pathogens) it requires qualified personnel for its use.

[0015] The invention proposes to remedy these limitations by describing a device capable of being manufactured at low cost and simple to implement. Statement of the invention

[0016] According to a first aspect of the invention, there is proposed a device for carrying out and reading the isothermal amplification reaction mediated by the loops on a sample to be analyzed received in a reaction tube, said device comprising an enclosure suitable for receiving said reaction tube.

[0017] The reaction tube is suitable for containing:

[0018] • the sample to be analyzed, • the reagents necessary for the LAMP reaction for DNA or RNA representative of the pathogen or a biological signature to be detected.

[0019] The enclosure comprises a well formed by:

[0020] • a first block insulating against heat, opaque to light radiation and pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the first block, • a second block which conducts heat and is opaque to light radiation, placed below the first block and also pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the second block, said hole in the second block extending the hole in the first block, • a third block, opaque to light radiation, placed below the second block and also pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the third block, said hole of the third block extending the hole of the second block, • a support arranged under the third block, opaque to light radiation, closing the hole opening onto the lower face of the third block, defining with the third block a lower cavity at the base of the second block, the lower cavity being suitable for receiving an end part of the reaction tube.

[0021] The enclosure further comprises a removable plug made of opaque material and closing the hole opening onto the upper face of the first block.

[0022] When the reaction tube is received in the well, the plug is arranged to prevent the entry of light radiation from outside the well through the hole that it closes, the second block is in thermal contact with the reaction tube, and the lower end portion of the reaction tube is received in the lower cavity.

[0023] By heat-insulating block, the present application designates a block having a thermal conductivity of less than 0.5 W / (mK).

[0024] By opacity, the present application relates to a material ensuring optical attenuation greater than 3 or 4 decades with respect to any external light radiation in the visible and near infrared range.

[0025] By heat-conducting block, the present application designates a block having a thermal conductivity lower than 150 W / (K. m).

[0026] The heat-conducting block can, for example, be made, without this being limiting, of copper or aluminum.

[0027] The production and reading device further comprises:

[0028] • a heating member in thermal contact with the second block and connected to control means, • a temperature sensor placed in thermal contact with the second

[0029]

[0030]

[0031]

[0032]

[0033]

[0034]

[0035]

[0036] block and / or the heating member, said temperature sensor being connected to control and measurement means, • a cooling member arranged to cool the second block, connected to control means, • a light source connected to control means, arranged to emit a light flux directed towards the base of the well, i.e. towards the base of the reaction tube when the reaction tube is received in the well, • a light flux sensor arranged to measure one (or more) wavelength(s) and positioned to receive a light flux from the base of the well, i.e. from the base of the reaction tube when the reaction tube is received in the well, • a computing unit. The computing unit is configured to: • measure the temperature of the second block using the temperature sensor • control the heating element via its control means, • control the cooling device (so as to lower the temperature of the second block), • control the luminous flux generated by the light source(s) • measure the intensity of the light received by the light flux sensor (re-emitted by the sample measured by one or more photometric sensors and determine whether the pathogen or biological signature to be detected is present or absent.) The heating element can, for example, be an electrical resistor. Controlling the heating element, for example the electrical power supplied by a resistor, has the effect of raising, and thus enabling regulation of, the temperature of the second block which is in thermal contact with the reaction tube. Controlling the cooling unit has the effect of lowering, and thus enabling regulation of, the temperature of the second block which is in thermal contact with the reaction tube. According to a preferred embodiment, the light sources consist of light-emitting diodes. The production and reading device according to the first aspect of the invention may further comprise communication means, of the light, wired or radio type, arranged to transmit information between the calculation unit and equipment external to the device and / or the user. Advantageously, a spectral filter may be functionally interposed between the light source and the reaction tube when the reaction tube is received in the well, i.e. between the light source and the sample to be analyzed when the tube reaction is received in the well. Thus, the emission spectrum of the light source can be shaped by the spectral filter so as to make its emission spectrum coincide with the excitation length(s) of the phosphor(s) used for the reaction.

[0037] Still advantageously, a spectral filter can be functionally interposed between the reaction tube and the light flux sensor, when the reaction tube is received in the well, i.e. between the sample to be analyzed and the light flux sensor, when the reaction tube is received in the well.

[0038] The light flux sensor may comprise one (or more) photodiode(s) associated with one (or more) bandpass filter(s). The technical effect is to make it possible to make the sensitivity spectrum of the filter coincide with the emission wavelength(s) of the luminophore(s) used for the reaction.

[0039] According to another preferred embodiment, the light flux sensor is of the spectrometer type.

[0040] Advantageously, the light flux sensor(s) are arranged on the longitudinal axis of the reaction tube, when the reaction tube is received in the well.

[0041] According to one possibility considered, the light fluxes from the light source(s) are emitted perpendicular to the axis and directed towards the base of the reaction tube. The technical effect is to prevent a significant portion of the excitation light flux from the sources from reaching the light flux sensor, the role of which is to measure the flux from the fluorescence emitted by the reaction base. This improves the signal-to-noise ratio.

[0042] According to one possibility, the light source is arranged in the removable cap.

[0043] According to a preferred embodiment, the processor, the control, measurement and communication means are grouped together on a single electronic circuit placed at the base of the third block.

[0044] According to a preferred embodiment, this electronic circuit also supports the light sources, the generated light being conducted near the reaction zone by means of light guides, followed or preceded by the spectral filtering means.

[0045] According to a preferred embodiment, the device comprises one or more contactors or touch keys making it possible to interact with the processor, and, for example, to start an amplification sequence.

[0046] The enclosure may comprise a plurality of wells. There are then as many light sensors as there are wells. The other means may optionally be shared or not. For example, the device according to the invention may comprise one or more cooling members, or as many as there are wells. The same applies to the heating member.

[0047] According to a second aspect of the invention, there is provided a system for producing and reading of the loop-mediated isothermal amplification reaction comprising:

[0048] • a sample to be analyzed received in a reaction tube, • a production and reading device according to the first aspect of the invention, or one or more of its improvements.

[0049] The sample is received in the well of said device.

[0050] According to a third aspect of the invention, there is provided a method for carrying out and reading the loop-mediated isothermal amplification reaction in a sample to be analyzed contained in a reaction tube, implementing a device according to the first aspect of the invention, or one or more of its improvements, comprising steps of triggering and controlling a loop-mediated isothermal DNA or RNA amplification reaction by generating one or more temperature cycles from the computing unit of said device.

[0051] There is provided, according to a fourth aspect of the invention, a computer program product comprising instructions which cause the device according to the first aspect of the invention, or one or more of its improvements, to carry out the loop-mediated isothermal amplification reaction steps on a sample to be analyzed received in a reaction tube in the well of said production and reading device.

[0052] According to a fifth aspect of the invention, there is provided a computer-readable medium on which the computer program according to the fourth aspect of the invention is recorded. Description of the figures

[0053] Other advantages and particularities of the invention will appear on reading the detailed description of implementations and embodiments which are in no way limiting, with regard to the appended drawings in which:

[0054] [Fig. 1] Figure 1 is a schematic illustration of a system according to the device described in patent US2020063197A1 which is prior art to the present application,

[0055] [Fig. 2] Figure 2 is an overall perspective view of an embodiment of a device according to the invention,

[0056] [fig.3] Figure 3 is a vertical section of the system illustrated in Figure 2,

[0057] [fig.4] Figure 4 is a perspective overview of the device shown in Figure 2, fitted with a stopper,

[0058] [fig.5] Figure 5 is a vertical section of the device illustrated in Figure 4,

[0059] [fig.6] Figure 6 is a schematic representation of different functions of a command processor,

[0060] [fig.7] Figure 7 is a perspective view of an electronic part of the system illustrated in Figure 2,

[0061] [fig.8] Figure 8 is a vertical section of a second embodiment of a device according to the invention,

[0062] [fig.9] Figure 9 is a vertical section of a third embodiment of a device according to the invention,

[0063] [fig.10] Figure 10 is a perspective overview of a fourth mode of production of a device according to the invention,

[0064] [fig.l 1] Figure 11 is a vertical section of the device illustrated in Figure 10,

[0065] [fig. 12] Figure 12 is a perspective overview of a fifth mode of production of a device according to the invention.

[0066] The embodiments described below being in no way limiting, it will be possible in particular to consider variants of the invention comprising only a selection of characteristics described, subsequently isolated from the other characteristics described, if this selection of characteristics is sufficient to confer a technical advantage or to differentiate the invention compared to the state of the prior art. This selection comprises at least one characteristic, preferably functional without structural details, or with only a part of the structural details if this part only is sufficient to confer a technical advantage or to differentiate the invention compared to the state of the prior art.

[0067] In the remainder of the description, elements having an identical structure or similar functions will be designated by the same references. Description of embodiment

[0068] A device 100 according to a first embodiment of the invention is now described with reference to Figures 1 to 7.

[0069] The device 100 has a substantially parallelepiped shape defining 6 faces, front, back, left, right, top and bottom.

[0070] The device 100 comprises an enclosure E suitable for receiving a reaction tube T, the enclosure comprising a well P.

[0071] The well P is formed sequentially, from its upper face to its lower face, by:

[0072] • a first block 1, opaque to light radiation and thermally insulating, pierced with a hole opening on the one hand onto an upper part and on the other hand onto a lower part of the first block and suitable for receiving an upper part of the reaction tube T, • a second block 2, opaque to light radiation and thermally conductive, placed below the first block 1 and also pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the second block, said hole of the second block extending the hole of the first block, • a third block 3, opaque to light radiation and thermally insulating, placed below the second block and also pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the third block, defining with the third block a lower cavity C at the base of the second block 2, the lower cavity being suitable for receiving an end part of the reaction tube.

[0073] The hole in the second block is of a shape complementary to a lower part of the reaction tube T. Thus, the hole in the second block forms an axial stop for the light tube. In the example shown, the hole in the second light block is of a substantially truncated cone shape.

[0074] When the reaction tube T is received in the well P, the lower end portion of the reaction tube is received in the lower cavity C.

[0075] The enclosure E further comprises a support 5 arranged under the third block, opaque to light radiation. The third block 3 rests on the support 5.

[0076] The support 5 is equipped with an electronic card Ce (Figure 6).

[0077] The device 100 further comprises a heating member 4 in thermal contact with the second block 2. According to the example shown, the heating member is in direct thermal contact with the second block 2, being attached to the second block.

[0078] According to the embodiment of the device 100, the heating member 4 is an electrical resistor. The electrical resistor is electrically connected to the electrical card Ce.

[0079] The device 100 further comprises a temperature sensor 6 arranged in thermal contact with the second block 2 and / or the heating member 4.

[0080] The device 100 further comprises a cooling member 7 for the second block 2, arranged to cool the second block 2, connected to control means (16). In the embodiment of the device 100, the cooling member is a fan directing a flow of air towards the second block 2 to enable it to be cooled.

[0081] In the example shown, block 2 is made of copper or aluminum. Its mass is minimized so as to also minimize the thermal inertia of the system. A size of 10 x 20 x 20 mm and a mass of around ten grams is generally used.

[0082] As illustrated in Figure 5, the reaction tube T generally has a volume of revolution. The volume of revolution has an upper part and a lower part. The upper part is generally formed of a cylinder with a circular base. The lower part is a conical or truncated part ending in a rounded base.

[0083] The reaction tube T comprises a reactive solution which is placed on one end 12 of the tube when the tube is presented vertically.

[0084] The device 100 further comprises:

[0085] • a light source 10 arranged to emit a light flux which is directed towards the base of the well P, that is to say towards the base of the reaction tube T when the reaction tube is received in the well P, • a light flux sensor 13, arranged to measure one (or more) wavelength(s) and positioned to receive a light flux coming from the base of the well.

[0086] In the example shown, the light source 10 is of the light-emitting diode type.

[0087] In the example shown, it can be noted that the light source 10 is mounted in a rear wall of the device according to the invention, in an accessible manner and to facilitate assembly. Still in the example shown, the light flux is directed perpendicular to the longitudinal axis of the reaction tube T.

[0088] In the example shown, the light flux sensor comprises 8 photodiodes.

[0089] In the example shown, the light flux sensor 13 is placed in the axis of the tube T and below its end 12.

[0090] Still according to the example shown, and optionally, a spectral filter 11 is optically interposed between the light source and the base of the well P, that is to say between the light source and the end 12 of the tube T, when the reaction tube is received in the well.

[0091] Still according to the example shown, and optionally, a spectral filter 14 is optically interposed between the base of the well P and the light flux sensor 13, that is to say between the end 12 of the tube T and the light flux sensor 13, when the reaction tube is received in the well.

[0092] Filter 11 allows the emission spectrum of the source to be adjusted to the excitation spectrum of the fluorophore. Filter 14 allows the sensitivity spectrum of the sensor to be adjusted to the emission spectrum of the fluorophore, thus optimizing the signal-to-noise ratio of the measurement.

[0093] As illustrated by Figure 4 and Figure 5, the device 100 further comprises a plug 8 opaque to light radiation and closing the hole opening onto the upper face of the first block 1. The plug prevents the entry of light radiation from outside the well P through the hole that it closes.

[0094] For this purpose, in the example shown, the plug 8 is provided with annular cylindrical walls and the block 1 is provided with an annular groove 30 cooperating with said annular cylindrical walls. Those skilled in the art can envisage other solutions.

[0095] Figures 6 and 7 show different functions of the electronic card Ce comprising a printed circuit supporting the components, in the form of a functional diagram and in perspective.

[0096] The functional diagram illustrates:

[0097] • the electrical resistance 4, mounted close to the thermal sensor 6, and arranged to be controlled in current by a control means 15. The resistance and the thermal sensor are thermally linked by any means known to those skilled in the art, such as a copper pad, not necessarily electrically connected, on the printed circuit, • the fan 7 arranged to be controlled by a control means 16, • a light source 10, or several, arranged to be controlled by a control means 17, • one or more light sensors 13, • a processor 18, arranged to control the control means, respectively 15, 16, 17 and receive the information coming from the sensors, respectively 6, and 13, • means 19 for supplying electrical energy, which provide the necessary current. These can come from a power supply external to the USB standard, well known to those skilled in the art, • wired (USB or others) or radio (WIFI, Bluetooth or others) communication means 20 which allow the user to communicate with the device, configure it and read the results, the processor 18 being arranged to receive and transmit information via the communication means.

[0098] Alternatively, or in addition, the communication means 20 may comprise usual manual interaction means, for example push buttons, light means, such as indicator lights or screens.

[0099] The heating member 4 is provided to raise the temperature of the second block 2, and thus allow it to be regulated, which triggers the LAMP reaction. In the case of an electrical resistor, this is obtained by varying the current in the resistor as a function of the temperature values ​​collected by the temperature sensor 6.

[0100] At the end of the isothermal reaction or several isothermal cycles, the number of DNA or RNA strands having multiplied sufficiently, reading by fluorescence is possible. The fan 7 is activated to cool the block 2 and stop the LAMP reaction.

[0101] The reading is carried out by activating the light source(s) 10 and collecting the signal re-emitted by the reactive solution by means of the light flux sensor 13.

[0102] Depending on the intensity of the re-emitted flow, and in comparison with a previously fixed threshold, it is possible to conclude whether the substance or organism to be detected is present or not.

[0103] Figure 8 is a second embodiment of a device 200 according to the invention, described only for its differences with the first embodiment.

[0104] The light source is arranged in the cavity C formed in the third block 3. This has the advantage of simplifying the implementation in the case, for example, of surface-mounted light sources.

[0105] Figure 9 is a third embodiment of a device 300 according to the invention, described only for its differences with the first embodiment.

[0106] The light source 10 is arranged in a part outside the enclosure C and the light flux is still directed towards the base of the reaction tube when the reaction tube is received in the well formed in the third block 3.

[0107] For this purpose, the device 300 further comprises a light guide block 20 which is mounted under the support 5 and receives the light flux emitted by the light source 10. The light guide block 20 is opaque to light radiation, and insulating with respect to heat. This has the advantage of being able, for example, to distribute the flux to several reaction tubes.

[0108] In order to be able to transmit the light flux towards the base of the reaction tube T, the support 5 is pierced to form a light conduit 52 in which one of the outputs of the light guide is inserted, thus bringing the light flux inside the cavity C.

[0109] This light guide can be made of transparent plastic material, for example of the PMMA type, for poly(methyl methacrylate).

[0110] Figure 10 and Figure 11 illustrate a fourth embodiment of a device 400 according to the invention, described only for its differences with the first embodiment.

[0111] The light source 10 is arranged in an outer part of the cap 8 and the light flux is still directed towards the base of the reaction tube T when the reaction tube is received in the well P formed in the third block 3. This has the advantage of being able to separate the light sources and sensors into two separate blocks, which can facilitate the production of a modular system.

[0112] Figure 12 illustrates a fifth embodiment of a device 500 according to the invention, described only for its differences with the first embodiment.

[0113] As previously, the device 500 comprises the first block 1, the second block 2, and the third block 3.

[0114] Unlike the first embodiment of the device 100, the first block 1, the second block 2, and the third block 3 comprise a plurality of wells P.

[0115] Each of the wells is associated with a stopper, receiving a light source 10.

[0116] Each of the caps receives a light source 10 arranged in a part ex inner part of the cap and the light flux is still directed towards the base of the reaction tube T when the reaction tube is received in the well formed in the third block 3.

[0117] Of course, the light source arrangements previously explained could be implemented.

[0118] Each of the wells is formed by holes in the first block 1, the second block 2, and the third block 3, as previously described.

[0119] The device 500 is provided with only one cooling member 7, designed and sized to cool the second block 2.

[0120] Unlike the first embodiment of the device 100, the device 500 is provided with two thermal sensors 6a, 6b.

[0121] Unlike the first embodiment of the device 100, the device 500 is provided with two heating members 4a, 4b.

[0122] Fluorophores are generally excited by a low wavelength (e.g. blue) to re-emit a signature at a higher wavelength (e.g. green). Two well-known fluorophores FAM and HEX exist and are both derivatives of fluorescein.

[0123] By being able to use two distinct pairs formed by a detector and a light source, it is then possible to distinguish two targets in the same reaction tube.

[0124] Suppose that pathogen #1 (PI) is associated with the fluorophore FAM, and pathogen #2 (P2) with the fluorophore HEX.

[0125] If we illuminate with light whose wavelength is approximately 480 nm, we can have in return:

[0126] • No response. Neither PI nor P2 are present. • The spectrum of HEX. P2 is present. • The spectrum of FAM. PI is present. • A mixture of both. PI and P2 are present.

[0127] There is therefore a great interest in having several detectors at several wavelengths.

[0128] Being able to illuminate / excite at multiple wavelengths to facilitate analysis of spectra is also useful, because by measuring for example at 480 nm and 540 nm excitation wavelength, we will see the spectra combine differently and increase the sensitivity of the test by matrix analysis of the results.

[0129] In addition, the various features, forms, variations and embodiments of the invention may be combined with each other in various combinations to the extent that they are not incompatible or mutually exclusive.

Claims

Claims

1. Device (100) for carrying out and reading the isothermal amplification reaction mediated by the loops on a sample to be analyzed received in a reaction tube (T), said device comprising: • an enclosure (E) suitable for receiving said reaction tube, said enclosure comprising a well (P) formed by: i. a first block (1) insulating against heat, opaque to light radiation and pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the first block, ii. a second block (2) conducting heat and opaque to light radiation, placed below the first block (1) and also pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the second block, said hole of the second block extending the hole of the first block, iii. a third block (3), opaque to light radiation, placed below the second block (2) and also pierced with a through hole opening on the one hand onto an upper part and on the other hand onto a lower part of the third block, said hole of the third block extending the hole of the second block, iv. a support (5) arranged under the third block, opaque to light radiation, closing the hole opening onto the lower face of the third block, defining with the third block a lower cavity (C) at the base of the second block, the lower cavity being suitable for receiving an end part of the reaction tube, • a removable plug (8) made of opaque material and closing the hole opening onto the upper face of the first block, when the reaction tube is received in the well, the plug prevents the entry of light radiation from outside the well through the hole that it closes and the lower end part of the reaction tube is received in the lower cavity, • a heating member (4) in thermal contact with the second block and connected to control means (15), • a temperature sensor (6) arranged in thermal contact with the second block and / or the heating member, said temperature sensor being connected to control and measuring means, • a cooling member (7) arranged to cool the second block, connected to control means (16), • a light source (10) connected to control means (17), arranged to emit a light flux directed towards the base of the reaction tube when the reaction tube is received in the well,• a light flux sensor (13) arranged to measure a wavelength and positioned to receive a light flux from the reaction tube when the reaction tube is received in the well, • a computing unit configured to: i. measure the temperature of the second block by means of the temperature sensor, ii. control the heating member via its control means, iii. control the cooling member, iv. control the light flux generated by the light source, v. measure the intensity of the light received by the light flux sensor.,

2. Production and reading device according to claim 1, further comprising communication means, of the light, wired or radio type, arranged to transmit information between the calculation unit and equipment external to the device.

3. A device for producing and reading according to claim 1 or 2, in which the light source is of the light-emitting diode type.

4. Device for producing and reading according to any one of the claims- preceding indications, wherein a spectral filter (11) is operatively interposed between the light source and the reaction tube (T), when the reaction tube is received in the well (P).

5. A device for producing and reading according to any one of the preceding claims, in which a spectral filter (14) is functionally interposed between the reaction tube (T) and the light flux sensor (13), when the reaction tube is received in the well (P).

6. Production and reading device according to any one of the preceding claims, in which the light flux sensor comprises a photodiode associated with a bandpass filter.

7. Production and reading device according to any one of claims 1 to 6, in which the light flux sensor is of the spectrometer type.

8. A device for producing and reading according to any one of the preceding claims, in which the light sensor is arranged on the longitudinal axis of the reaction tube when the reaction tube is received in the well (P).

9. System for carrying out and reading the loop-mediated isothermal amplification reaction comprising: • a sample to be analyzed received in a reaction tube (T), • a carrying out and reading device according to any one of the preceding claims, in which the sample is received in the well (P) of said device.

10. Method for carrying out and reading the loop-mediated isothermal amplification reaction in a sample to be analyzed contained in a reaction tube (T), implementing a device according to claim 1, comprising steps of triggering and controlling a loop-mediated isothermal DNA or RNA amplification reaction by generating one or more temperature cycles from the computing unit of said device and determining whether a pathogen or a biological signature to be detected is present or absent.

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