Mobile genetic analysis laboratory

Description

[0001] The present invention relates to a mobile laboratory for the serial analysis of nucleic acids from a biological sample, in particular in the field of forensics.

[0002] It concerns the technical field of mobile laboratories that can be quickly transported to a location, in particular crime scenes (e.g., crime scenes) for the purpose of rapid detection and identification, in particular of the identity of people, from a biological sample.

[0003] Typically, a team of specialists collects samples (blood, bone marrow, buccal cells, degraded or non-degraded human biological tissue) at a crime scene. These samples are then packaged and transported by specialized vehicle to a genetic analysis laboratory to identify the person associated with the sample. Once the identification is made, information on the identity of the person(s) thus identified is transmitted to the competent authorities who take the appropriate measures.

[0004] The specialized vehicles transporting the samples are at best refrigerated vehicles or at least vehicles equipped with a cooler, as no diagnosis can be carried out inside the vehicle itself. In addition, there are very few centers with a genetic analysis laboratory accredited for the identification of individuals, so the time taken to transport the samples from the crime scene to the analysis laboratory can take several hours or even several days depending on the size of the territory, the conditions of access to the crime scene, its location in relation to the nearest analysis laboratory and the means of transport available, as for example in the Germanwings air crash that occurred on March 24, 2015, where a plane crashed in the heart of the French Alps.This time lapse between the collection of samples and the transmission of information on the identity of the persons associated with the samples to the competent authorities is therefore not optimal, which is highly detrimental in particularly sensitive situations where the time required to obtain analysis results is a crucial parameter.

[0005] The document A. HOPWOOD et al. "Forensic response vehicle: Rapid analysis of evidence at the scene of a crime", INTERNATIONAL CONGRESS SERIES., vol. 1288, April 1, 2006 pages 639-641 discloses a mobile laboratory for the serial analysis of nucleic acids. However, the mobile laboratory described in this document does not allow for real energy autonomy, and lacks prevention regarding sample contamination.

[0006] To date, so-called mobile laboratory models are available on the market, but their activity is dedicated to the detection of pathogens (virology, bacteriology), chemical substances, or more generally military field laboratories for the purposes of medical care and simple biological analyses. Examples include the mobile laboratories described in the documents: Giorgio MARI et al.: “CRBN mobile laboratories in Italy,” SPIE - INTERNATIONAL SOCIETY FOR OPTICAL ENGINEERING. PROCEEDINGS, vol. 7304, May 1, 2009, pages 73040W1 to 73040W12, and AGILENT “Agilent mobile laboratory for chemical and biological defense,” April 30, 2003, pages 1-8.However, none of these mobile laboratories can carry out complete genetic analyses, particularly human ones (DNA or RNA), from the sampling to the determination with certainty of the identity of the person associated with the sample, from the sampling location, while respecting the constraints linked to the risks of contamination and therefore of incorrect identification of the person specific to the field of forensics.

[0007] The constraints encountered for the implementation of such a mobile laboratory are the environmental constraints linked to the risks of DNA contamination as well as to the traditional equipment necessary for the amplification of nucleic acids and in particular the equipment necessary for sequencing which are not designed to be moved from their initial installation site. These constraints are fundamentally different from those of a laboratory which would be designed to identify pathogenic agents in an NRBC (nuclear, radiological, biological and chemical weapon) framework, where the aim would be to protect the operator from the external environment and the danger of the sample, whereas in the case of the mobile laboratory according to the invention, the aim is to protect the sample against contamination by the operator and the external / analysis environment.

[0008] The technical problem to be solved was therefore to propose a transportable laboratory as close as possible to a crime scene, configured with all the traditional equipment necessary and adapted to enable identification of people by genetic analysis from a sample, in analysis conditions at least as reliable as in a conventional laboratory so as not to be able to call into question the reliability of the results. This problem must be solved while allowing easy implementation by personnel traditionally carrying out these operations in a conventional genetic analysis laboratory, without the need for additional training, and at an acceptable cost, in particular for occasional use by police or armed forces where the cost is covered by the community.Finally, it is desirable that this mobile laboratory can be transported as close as possible to any type of crime scene, regardless of its geographical location.

[0009] The Applicant solved this problem by developing a mobile laboratory of simple design, comprising the genetic analysis equipment routinely used in a traditional laboratory and adapted for the mobile laboratory when it is necessary to move it, not requiring additional training of staff because it uses the same equipment and the same protocols and is quickly operational at the crime scene, and capable of being easily transported by road, but also by sea, rail or air.

[0010] The said mobile laboratory has a configuration which allows the principle of "forward movement" of sample processing to be respected, namely: a first zone in which the sample to be analyzed is inserted into the laboratory and enters the analytical zone where the technical operations preparatory to gene amplification by PCR are carried out; a second zone where both gene amplification by PCR in the thermocycler (22), contacting of the amplification products with the sequencing reaction mixture, and sequencing are carried out.

[0011] Another aim of the invention is to provide a mobile laboratory in a vehicle, moving autonomously on the road.

[0012] The mobile laboratory according to the invention allows for the serial analysis of nucleic acids outside the conventional genetics laboratory, thus providing a mass analysis capacity as close as possible to the sampling scene. Using this collection and analysis method, the time between the installation of the laboratory on site, the collection of the biological trace and the determination of the genetic profile is less than three hours for the first twenty-one samples analyzed. The analysis system then makes it possible to generate at least twenty-one additional results every twenty to twenty-five minutes. The time taken to obtain results is thus considerably reduced compared to the time required for transport and analysis in a conventional laboratory, while retaining the advantages of producing serial analyses of several dozen samples.

[0013] This mobile laboratory is designed to intervene quickly at crime scenes, for example in the context of a terrorist attack, and to carry out a formal identification of victims or suspects of the crime directly in the area concerned, in order to inform the competent authorities in real time so that they can quickly take the necessary decisions regarding the measures to be put in place (confirmation or denial of the identity of the victim(s) to the families, arrest warrant for a suspect, call for witnesses, etc.).

[0014] Finally, the use, for the collection of the sample to be analyzed in the mobile laboratory according to the invention, of a tool for collecting biological material comprising a rod having at least one end a dry absorbent body comprising surfactants and denaturing agents, makes it possible to eliminate the DNA extraction step and therefore to reduce the cost of labor, equipment and laboratory consumables necessary for genetic analysis, while reducing the risks of contamination of the sample.

[0015] A first object of the present invention relates to a mobile laboratory (1) formed of a transportable mobile container defining a basic room, the interior space of said room being isolated from the outside by means of at least one closing system (8), said room comprising: o at least two spaces distinct from each other i. The first space (2) called “pre-PCR” suitable for carrying out a pre-amplification step and comprising: A mixture of PCR amplification reaction of nucleic acids A container ii.a second space (3) called "post-PCR" suitable for carrying out a step of amplification of nucleic acids by PCR, sequencing and genotyping comprising: A PCR analysis device (22) A sequencer (20) A sequencing reaction mixture A container ∘ an alternating opening airlock system (4a) between the exterior and the pre-PCR zone (2) of a size suitable for the passage of a sample and not of a human being, said alternating opening airlock system (4a) allowing the transmission of samples between the exterior and the pre-PCR space (2); ∘ a closing system (4b) between the two spaces (2) and (3) said closing system (4b) making the two spaces (2) and (3) sealed from each other and allowing the transmission of samples between the two spaces (2) and (3); o an autonomous current inverter associated with batteries.Said mobile laboratory being adapted to the genetic analysis and identification of an individual from a biological sample directly at the place where said sample was taken.

[0016] By closing system, we mean according to the present invention a door, preferably sliding, and allowing sealing between the two spaces that it separates once closed.

[0017] According to one embodiment of the invention, the closure system (8) separates the exterior of the mobile laboratory from the post-PCR space. The user of the mobile laboratory according to the invention then enters the interior of said laboratory via the closure system (8) into the post-PCR space, closes the closure system (8), then enters the pre-PCR space via the closure system (4). He then closes the closure system (4) in order to ensure the seal between the 2 spaces.

[0018] Preferably, the invention may comprise two closure systems (8), one between the exterior of the mobile laboratory (1) and the pre-PCR space (2), and the other closure system between the exterior of the mobile laboratory (1) and the post-PCR space (3). This configuration is presented to the figures 1 And 2 .

[0019] Alternatively, the invention may comprise an airlock, accessed by the closure system (8), which allows a user of the mobile laboratory to access the interior of the mobile laboratory from the outside, said airlock comprising two additional closure systems, one allowing access to the pre-PCR space (2), and the other to the post-PCR space (3), these two closure systems not being able to be opened together, and both being able to open only when the closure system (8) between the outside and the airlock is closed, in order to prevent any contamination.

[0020] Preferably, said closing system (8) is an airlock, with alternating opening. In a particularly preferred manner, the mobile laboratory according to the invention comprises two airlocks, one between the exterior of the mobile laboratory (1) and the pre-PCR space (2), and the other between the exterior of the mobile laboratory (1) and the post-PCR space (3). Said airlocks constitute a dressing area between the exterior and the interior of each analytical zone in which the operator can remove his clothing and put on clothing dedicated to laboratory activities.

[0021] Preferably, the closure system (4) between the pre-PCR space and the post-PCR space is an alternating opening airlock system (4) between the two spaces (2) and (3) of a size suitable for the passage of a sample and not of a human being, said alternating opening airlock system (4) making the two spaces (2) and (3) sealed from each other and allowing sample communication between the two spaces (2) and (3). Preferably, said airlock does not allow the passage of a human being.

[0022] According to the invention, the airlock system with alternating opening (4a) between the exterior and the pre-PCR zone (2) of a size suitable for the passage of a sample and not of a human being, allowing the transmission of samples between the exterior and the pre-PCR space (2), is not of a size to allow the passage of a human being. Indeed, the user of the mobile laboratory enters the interior of said laboratory via the closing system (8). Said system makes it possible to respect the principle of "forward movement" of the samples (and of the user of the mobile laboratory) and therefore to combat the contamination of the samples as best as possible.In particular, the absence of such an alternating opening airlock system (4a), which would require samples to be passed through the closing system (8), is not ideal in the forward movement process, which must differentiate the forward movement of samples from that of the users of the mobile laboratory in order to best combat DNA contamination of the samples. PRE-PCR AREA

[0023] The nucleic acid PCR amplification reaction mixture as present in the pre-PCR space of the mobile laboratory according to the present invention refers to a particular combination of reagents that are necessary to carry out a PCR amplification reaction. The amplification reaction mixture used in each case and the conditions applied to carry out the amplification are well known to those skilled in the art. For example, the amplification reaction mixture may comprise one or more sets of primers to ensure that a particular target nucleic acid is subjected to the amplification reaction, as well as enzymes, nucleotides, probes, salts and buffers that may also be necessary to carry out amplification. The amplification reaction medium can typically amplify STR (Short Tandem Repeat) type DNA fragments such as in the Identifiler plus and Globalfiler kits (Lifetechnologies)

[0024] Said nucleic acid PCR amplification reaction mixture is added in an amount sufficient to saturate the absorbent body of the device according to the invention. It is typically added in an amount of 7.5 to 25 microliters, preferably 12.5 to 15, more preferably 12.5 microliters.

[0025] Advantageously, the Pre-PCR space also includes at least one workbench, one of which covers a refrigerated chamber from +4°C to -20°C (21), a laminar flow hood, at least one container and the small basic equipment (vortex, centrifuge, plate sealer, micropipettes).

[0026] Preferably, the container consists of the well(s) of a plate or strip, for example 8-well, 96-well or 384-well, and is ready for use for PCR. Thus, preferably, the container is pre-filled with the nucleic acid PCR amplification reaction mixture. Advantageously, said container also comprises the amplification reaction controls, pre-filled in the container and already added to the ready-to-use amplification mixture.

[0027] By container is meant a container for the absorbent body in order to carry out the contacting of the absorbent body of a biological material collection device with the nucleic acid PCR amplification reaction mixture and the PCR as well as the contacting of the amplification products with the sequencing reaction mixture. For example, according to the present invention, it is intended to denote a container with a capacity of 10 to 350 microliters, or a combination of containers with a capacity of 10 to 200 microliters such as a 96-well PCR microplate, or a 384-well PCR microplate. Preferably, the container is a 96-well PCR microplate of 100 or 200 microliters.

[0028] In one embodiment of the invention, the pre-PCR space is a glove box. According to the present invention, the term "glove box" means a sealed enclosure that allows manipulations in a particular atmosphere. Shoulder-length gloves, attached to one of the walls, allow access to the interior without ceasing the confinement. The user places his hands in the gloves and then sees his manipulations through the transparent wall. POST-PCR SPACE

[0029] Advantageously, the Post-PCR space also comprises at least one workbench, one of which covers a refrigerated chamber from +4°C to -20°C (21), at least one container and the small basic equipment (vortex, centrifuge, micropipettes, plate sealer). The container is advantageously as described above.

[0030] Said nucleic acid sequencing reaction mixture as present in the post-PCR space of the mobile laboratory according to the present invention refers to a particular combination of reagents which are necessary to carry out sequencing of amplified DNA fragments. The sequencing reaction mixture used in each case and the conditions applied to carry out sequencing by the electrophoresis technique are well known to those skilled in the art. For example, the sequencing reaction mixture may comprise formamide as well as a set of DNA fragments of known molecular weight, commonly referred to as a size standard by those skilled in the art. Depending on the size assigned to them, each amplified DNA fragment is identified by comparison with a set of reference DNA fragments commonly referred to as an allelic ladder by those skilled in the art.

[0031] The container is preferably pre-filled with the sequencing reaction mixture of the nucleic acid PCR amplification products. Advantageously, said container may also be pre-filled, in addition to the sequencing reaction mixture, with a set of reference DNA fragments, commonly called an allelic ladder, allowing the identification of the amplification products.

[0032] According to another advantageous characteristic of the invention, a PCR (Polymerase Chain Reaction) analysis device is arranged in the post-PCR space so that the personnel can select and multiply a gene or an identifiable fraction of DNA or RNA chain in order to identify the person associated with the biological sample, in less than two hours.

[0033] This PCR analysis device (22) is advantageously a thermal cycler. It can be any thermal cycler that is commercially available for equipping a traditional genetic analysis laboratory. Preferably, it has an electrical outlet that meets international standards, such as a type A and B or type C, E and F outlet. Thus, it can be, for example, a thermal cycler of the GeneAmp PCR system 9700 or Veriti models marketed by the company Thermofisher.

[0034] Advantageously, a crate for facilitating the transport of said PCR analysis device (22) may be used.

[0035] Comparative PCR DNA amplification performance tests were carried out with the same thermal cycler (Veriti model from Thermofisher) in a conventional laboratory and in the mobile laboratory after having traveled 800 km, without any particular calibration test and provided the same results.

[0036] The genotyping of the PCR products is then carried out using a sequencer analyzer device (20), called a "sequencer". A gene sequencer (20) is a device capable of automating the DNA sequencing operation. It can be any sequencer (20) that is commercially available for equipping a traditional genetic analysis laboratory. Preferably, it has a standard electrical outlet. Thus, it can be, for example, a sequencer (20) of the ABI 3500 XL model marketed by the company Thermofisher.

[0037] The sequencer (20) is generally a large instrument that can weigh around a hundred kilos and is sedentary and uses sensitive optical systems that are likely to become deregulated over time. It is therefore a critical instrument to transport. For this purpose, advantageously in the context of the present invention, the optics of the sequencer (20) can be subject to greater fixation, namely that the fixation system of the optics of the sequencer (20) has been tightened compared to traditional laboratory use which is traditionally a sedentary use of a tool that is not intended to be moved. This tightening may only be checked during the annual maintenance visit of the instrument.

[0038] There are many sequencer models and it is not possible to define a standard tightening that could be applied to every sequencer on the market, nor those that could be marketed in the future. Indeed, each sequencer has its own optical system and its own original tightening force.

[0039] It is the person skilled in the art who is able to evaluate, for the sequencer model that he wishes to use in the context of the invention, that is to say in a mobile laboratory with a transport step likely to disrupt the optical system of said sequencer, whether it is necessary to tighten said optical system and to what extent. He may for this and conventionally use the information provided by the manufacturer of the sequencer and may, in accordance with his general knowledge, carry out a calibration, for example by testing the original tightening and then different re-tightenings of the optical system of the sequencer so that it is transported, or subjected to conditions similar to transport (for example by placing said sequencer on a plate subjected to controlled back and forth, circular, vibration and oscillation movements at a magnitude between 500 microns up to 1 cm, at a frequency of 5 to 50 Hz), without the optical systems becoming disrupted.

[0040] Such calibration on capillary electrophoresis systems can for example be carried out using DNA fragments of known molecular weights labeled with fluorescent molecules so that they can be excited by the laser using the sequencer's optical system and then detected by the camera.

[0041] If the optical system is out of adjustment then the DNA fragments will not be detected by the camera or will be at molecular weights different from those expected.

[0042] Tightening the optical system can be done conventionally using a torque tool such as a screwdriver.

[0043] Preferably, prior to tightening the optics, all threads of the sequencer's optical system can be treated with a weak locking product such as LOCTITE ®< 221 ™< which is a polymer designed for locking and sealing threaded assemblies. Its function is to prevent loosening or leaks due to shock and vibration.

[0044] As an example, the tightening of the optical system of the Thermofisher sequencer, model 3500 XL, will be described below. Prior to tightening the optics, all threads are treated with a weak locking agent LOCTITE ®< 221 ™< .

[0045] M3 type hexagonal nuts, 5.5 mm dimension, whose initial tightening torque value does not exceed 20 cN.M are tightened using a torque screwdriver until a tightening torque of 35 cN.M (centiNewton-meter) is obtained with their bolt.

[0046] M2.5 mm hex socket head bolt heads with an initial tightening torque value not exceeding 20 cN.M are tightened using a torque screwdriver until a tightening torque of 40 cN.M is obtained with their bolt.

[0047] It is observed on average that the tightening torque of the bolt heads with their bolt of the optical systems of several sequencers available on the market does not exceed 20 cN.M

[0048] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that said sequencer (20) is arranged in an anti-vibration transport device. Said anti-vibration transport device is preferably a transport case made to measure in relation to said sequencer (20) and lined with a viscoelastic vibration-damping material such as a damping polymer foam. Advantageously, said viscoelastic material is a polyethylene foam, such as the non-crosslinked polyethylene foam ETHAFOAM 220 and ETHAFOAM 400 marketed by Dow Chemical, which are resilient and robust closed-cell foams with an average density of 35kg / m3 and 105kg / m3 respectively.

[0049] Another anti-vibration transport device may include VIBCABLE type cable dampers marketed by PAULSTRA.

[0050] Preferably, after each movement of the mobile laboratory and before using the sequencer (20) for genetic analysis, a calibration procedure is carried out. This is the same procedure as that carried out when installing the sequencer (20) upon its initial delivery to a traditional laboratory.

[0051] In a particular embodiment of the invention, said mobile laboratory may comprise several thermocyclers and / or several sequencers in order to increase the number of samples which can be identified over a given period of time.

[0052] Advantageously, the computer cockpit has also been changed compared to the standard version: a laptop replaces the desktop computer.

[0053] Comparative performance tests carried out in a conventional laboratory and in a mobile laboratory with a sequencer (20) of the ABI 3500 XL model marketed by the company Thermofisher, provided the same results.

[0054] Advantageously, the mobile laboratory according to the invention comprises a lifting table for transporting the transport boxes of the thermocycler (22) and / or the sequencer (20) inside and then outside the mobile laboratory.

[0055] In order to avoid human DNA contamination which is a critical factor in forensic DNA analysis, the sequencer (20) and the thermal cycler (22) are inserted and removed from the mobile laboratory, via an instrument pass-through (6) with alternating opening consisting of a "rolling shutter" or a 'swinging wall' accessible from outside the vehicle and a rolling shutter accessible from inside the laboratory in the post-PCR analytical area, making it possible to protect the laboratory area from the outside environment and from the entry of individuals into this area.Preferably, the mobile laboratory according to the invention comprises at least one instrument pass-through (6) and more preferably one instrument pass-through (6) per device, consisting of a rolling shutter or a pivoting wall each giving access from the outside to an airlock opening alternately with the inside of said laboratory, advantageously a rolling shutter (6s) or a pivoting wall for the passage of the sequencer and giving access to a sequencer airlock, and a rolling shutter (6t) or a pivoting wall for the passage of the thermocycler, and giving access to a thermocycler airlock;

[0056] Typically, the base of the instrument pass-through (6s) of the sequencer (20) can be raised by a mechanical action initiated from inside the post-PCR area in order to constitute a work surface at standard height (85-90cm from the floor of the vehicle). Once in the raised position, the storage area of ​​the cover of the transport box of the sequencer (20) is located under the base of the instrument pass-through (6s) once it is in the raised position. An equivalent system of instrument pass-through (6t) is developed for the thermocycler (22) without the need for a lifting base. An equivalent system of rolling shutter for the generator is developed.

[0057] The instrument pass-throughs have hooks for securing the instrument transport boxes to ensure they remain in place when the vehicle is moving.

[0058] In operation, the instruments remain in the base of their transport case.

[0059] This means that critical materials can be inserted and removed from the laboratory without having to enter the analytical area, thus protecting these areas from any dirt and contamination from the external environment.

[0060] Preferably according to the invention, the base of the instrument holder can also slide towards the outside of the mobile laboratory in order to facilitate the positioning of the instruments in the instrument holder. AUTONOMOUS ELECTRICAL DEVICE

[0061] By device guaranteeing an autonomous, continuous and backed-up power supply, is meant according to the present invention an autonomous current inverter associated with batteries making it possible to prevent micro-cuts in the current and to provide the energy necessary to operate without current or a generator (7). Advantageously, the mobile laboratory according to the invention however also comprises a generator (7), and a 12v / 220V converter electrical panel (or 110V depending on the country of use of the mobile laboratory). The autonomous inverter delivers a voltage with a frequency that is either fixed or adjustable by the user. It does not always need an electrical network to operate; for example, a travel converter that is plugged into the cigarette lighter socket of a car uses the vehicle's 12V DC to generate 120 or 230V, AC at 50 or 60 Hz;

[0062] Preferably, the device guaranteeing an autonomous, continuous and emergency power supply is stored in the trunk (9) of the vehicle. In the figure 1 location (5) corresponds to the electrical panel of the laboratory area (12v / 220v converter) and the location of the inverter and associated batteries.

[0063] The said device guarantees uninterrupted power supply during the entire PCR amplification and sequencing process.

[0064] The laboratory in accordance with the invention can thus operate completely autonomously at crime scenes. AIR CONDITIONING

[0065] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it comprises an air conditioning device (13). Said device makes it possible to control the ambient temperature conditions. Indeed, environmental conditions having an impact on the electrophoresis conditions during genotyping with the sequencer, it is important to be able to control the ambient temperature to certify that the analyses are carried out in a temperature range meeting the specifications of the sequencer.

[0066] Preferably, the laboratory according to the invention has an independent air conditioning device (13) for each pre-PCR and post-PCR space. Advantageously, said device also makes it possible to control the ambient humidity conditions and limit the contamination of the spaces by dust.

[0067] In genetic analyses, it is desirable to maintain constant temperature conditions, preferably around 21°C, in the pre-PCR and post-PCR areas.

[0068] Advantageously, said mobile laboratory according to the present invention also includes a continuous temperature recorder allowing the conditions of use of the instruments to be traced. A refrigerated chamber +4°C / -20°C is also present in each zone. Their temperature is also traced. These chambers can operate when the laboratory is mobile, thus ensuring continuity in the control of the cold chain which guarantees the preservation of reagents and consumables. TELEMATICS CONNECTION AND COMPUTER NETWORK

[0069] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it comprises a computer device, such as at least one laptop, an internal computer network and a secure telematics link. Said internal computer network allows data to be exchanged within the internal perimeter of the mobile laboratory. Said telematics link allows information and in particular the genetic profiles thus determined to be transmitted to any point on the globe, for example to an expert or to national or international databases.

[0070] The telematics link also allows video to be transmitted via cameras installed in each of the technical areas, which makes it possible to record all the technical operations carried out but also to view all these technical operations in real time from outside the mobile laboratory and remotely.

[0071] Such a telematics link is, for example, a means of wireless internet connection such as a 4G key, or a satellite link, associated with a means of encryption or ciphering.

[0072] Preferably, the mobile laboratory according to the present invention comprises an air conditioning device (13), a secure telematics link, and a local computer network. CLOSING SYSTEM and SAS

[0073] By closing system, we mean according to the present invention a door, preferably sliding, and allowing sealing between the two spaces that it separates once closed.

[0074] According to one embodiment of the invention, the closure system (8) separates the exterior of the mobile laboratory from the post-PCR space. The user of the mobile laboratory according to the invention then enters the interior of said laboratory via the closure system (8) into the post-PCR space, closes the closure system (8), then enters the pre-PCR space via the closure system (4). He then closes the closure system (4) in order to ensure the seal between the 2 spaces. According to another embodiment, the user of the mobile laboratory according to the invention can enter the interior of said laboratory via the closure system (8) into the pre-PCR space and then access the post-PCR space via the closure system (4).

[0075] Preferably, the invention may comprise two closure systems (8), one between the exterior of the mobile laboratory (1) and the pre-PCR space (2), and the other closure system between the exterior of the mobile laboratory (1) and the post-PCR space (3). This configuration is presented to the figures 1 And 2 .

[0076] Preferably, said closing system (8) is an airlock, with alternating opening. In a particularly preferred manner, the mobile laboratory according to the invention comprises two airlocks, one between the exterior of the mobile laboratory (1) and the pre-PCR space (2), and the other closing system between the exterior of the mobile laboratory (1) and the post-PCR space (3). Said airlocks constitute a dressing area between the exterior and the interior of each analytical zone in which the operator can remove his clothing and put on clothing dedicated to laboratory activities.

[0077] Preferably, the closure system (4) between the pre-PCR space and the post-PCR space is an alternating opening airlock system (4) between the two spaces (2) and (3) of a size suitable for the passage of a sample and not of a human being, said alternating opening airlock system (4) making the two spaces (2) and (3) sealed from each other and allowing the transmission of samples between the two spaces (2) and (3).

[0078] Preferably according to the invention, each airlock or closing system in the mobile laboratory is equipped with an intercom allowing communication with the outside or other spaces of the mobile laboratory.

[0079] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it further comprises an additional sampling space (11) used for taking biological samples and which may comprise, for example, sampling tables such as autopsy tables.

[0080] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it further comprises a sample storage space used for storing the biological samples taken and which may comprise, for example, coolers.

[0081] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it further comprises an interpretation zone used for the interpretation of the results obtained after sequencing and which may comprise, for example, computers and a telematics link.

[0082] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it further comprises a meeting area making it possible to plan the analyses and communicate the results obtained.

[0083] In a particularly preferred manner according to the invention, the mobile laboratory according to the invention is characterized in that it comprises an additional mobile space (11) for sampling and / or storage of samples, and / or an interpretation area, and / or a meeting area.

[0084] According to an advantageous characteristic of the invention, the mobile laboratory comprises at least one additional space (11) which can be mounted on site at the scene of the offense and forming at least one annex in which said additional sampling space, said sample storage space, said interpretation area or said meeting area can be installed. In the maximum configuration where the mobile laboratory has an additional sampling space, a sample storage space, an interpretation area and a meeting area, the total surface area of ​​the mobile laboratory and its annexes is approximately 40 m2 which is sufficient space to work in good conditions.

[0085] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that said complementary spaces (11) are mobile. Preferably, said complementary spaces are inflatable and / or deployable. In particular, they may be walls deployable from the mobile laboratory.

[0086] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that the small basic materials, the reagents and consumables necessary for the analyses, the PCR analysis device (22), the sequencer (20) and the autonomous current inverter associated with batteries are mobile and can be removed from the mobile laboratory after each use of said laboratory. This system makes it possible to optimize the use of the instruments on a daily basis in a mobile laboratory according to the invention but also in a traditional laboratory, and to transfer said instruments into the mobile laboratory only when necessary, and without it being necessary to have expensive equipment, in particular a PCR analysis device (22) and a sequencer (20) for each of the mobile and traditional laboratories. VEHICLE

[0087] Preferably according to the invention, the laboratory is arranged inside a vehicle.

[0088] By vehicle, according to the present invention, is meant a motorized mobile vehicle, typically a van or a bus. Preferably, said motorized vehicle is driven with a driving license for passenger vehicles (License B in France). This makes it possible, in the event of an emergency intervention, to expand the number of people who can bring said mobile laboratory to an intervention zone. Advantageously, according to the invention, the mobile laboratory is arranged directly inside said mobile vehicle.

[0089] Preferably, said vehicle has at least one audible and visual warning device (14) enabling it to be signalled to other users when it is en route to the scene of the offence and to facilitate the transport of the vehicle as quickly as possible and in complete safety.

[0090] A storage area for said lifting table is dedicated in the trunk (9) of the mobile laboratory vehicle.

[0091] The mobile laboratory according to the invention has external electrical sockets (15) allowing connection to the electrical network, in particular if a generator is not used, and internal electrical sockets in the analytical area, as well as lighting, in particular if said laboratory does not include windows.

[0092] Advantageously, the mobile laboratory according to the invention comprises a system for stabilizing and horizontally leveling the vehicle when stationary, which could be useful depending on the configuration of the terrain at the projection site.

[0093] An area for storing service weapons of the police or armed forces can also be provided in the mobile laboratory.

[0094] In a particularly preferred manner according to the invention, the mobile laboratory can be moved by any means, road, air, rail or sea. In particular, the mobile laboratory can be moved by air by being loaded onto a military aircraft, for example of the C-160 TRANSALL type. COLLECTION DEVICE WITHOUT EXTRACTION STEP

[0095] Preferably according to the invention, the mobile laboratory according to the invention is characterized in that it comprises a device for collecting biological material comprising a rod having at least one end a dry absorbent body comprising surfactants and denaturing agents. Preferably, said absorbent body has a surface area of ​​between approximately 1 and approximately 3.14 mm 2<

[0096] According to the present invention, the term “biological material” means any material derived from a living being, which may be in the form of molecules, organelles, fluids, cellular fragments or cells, and present on a biological or non-biological support. Preferably, said biological material comprises nucleic acids, preferably DNA, in a form allowing their amplification by PCR (Polymerase Chain Reaction) and their analysis and / or identification.Said biological material is advantageously selected from the group consisting of physiological body fluids, cells or cell suspensions of humans or animals, in particular a contact trace, blood, bone marrow, buccal cells, degraded or non-degraded human biological tissue or cell suspensions of plants; liquid products, extracts or suspensions of bacteria, fungi, plasmids, parasites, or viruses; liquid extracts or homogenates of human or animal body tissues; media resulting from a synthesis of nucleic acids; and mixtures of chemically or biochemically synthesized nucleic acids. In a particularly preferred manner, the biological material according to the present invention is in particular a contact trace, blood, bone marrow, buccal cells, degraded or non-degraded human biological tissue.

[0097] According to the present invention, nucleic acids are understood to mean DNA and RNA. Preferably, according to the present invention, they are understood to mean DNA.

[0098] According to the present invention, the term “absorbent body” is understood to mean a dry solid support composed of an absorbent material, natural or synthetic. Preferably, said absorbent body is chosen from the group consisting of cellulose, nitrocellulose, cotton, flocked synthetic fibers, for example flocked nylon fibers, hydrophilic polymers, fiberglass, polytetrafluoroethylene, polyester, nylon, rayon, cotton and a porous ceramic. More preferably, said absorbent body is chosen from cellulose and flocked synthetic fibers. More preferably, said absorbent body is composed of flocked synthetic fibers, preferably flocked nylon fibers, preferably directly on the stem. Advantageously, the fibers are glued to the stem. Preferably, the fibers measure between 1 and 1.5 mm in length and between 0.01 and 0.5 mm in diameter.This configuration allows for the best results in terms of collection of biological material and release of nucleic acids from said biological material, combined with simplicity of manufacture and use.

[0099] Due to its miniature surface area of ​​around 1 to 3.14 mm 2< , the absorbent body is again of major economic interest because this characteristic makes it possible to carry out the amplification of nucleic acids in a reaction volume reduced by 50% compared to the supplier's recommendations and therefore to considerably reduce the cost of analyzing a large number of samples. Indeed, the tests carried out with the Identifiler plus and Globalfiler gene amplification kits have made it possible, for example, to demonstrate the effectiveness of the device for a PCR reaction volume of 25µl as recommended by the supplier, but also in a reaction volume reduced by half, i.e. 12.5µl. It has thus been demonstrated that such a small surface area of ​​the absorbent body makes it possible to collect a sufficient quantity of nucleic acids to obtain a genetic profile.The absorbent body is pre-treated with denaturing agents and surfactants during its design, thus allowing in situ lysis of cells. It is therefore not necessary to apply a specific humidification buffer to this absorbent body for cell lysis before collection.

[0100] Surfactants and denaturants are understood to mean agents for cell lysis, protein denaturation and nucleic acid extraction. Surfactants and denaturants may be selected from a list including uric acid or urate salt, tris, SDS guanidine salt, preferably guanidine thiocyanate, Triton X100, Tween 20, CHAPS, CHAPS-O, octyl glucoside, octyl thioglucoside and combinations thereof.

[0101] Preferably according to the invention, the surfactants and denaturants are a combination of uric acid or urate salt, tris, and SDS. Preferably, the absorbent body further comprises EDTA. In particular, the absorbent body comprises a mixture per cm2 of 2 micromoles of uric acid, 8 micromoles of Tris, and 1 mg of SDS. Preferably, the absorbent body further comprises 0.5 micromoles of EDTA.

[0102] According to a variant of the invention the surfactant and denaturing agents may be a combination of guanidine salt, preferably guanidine thiocyanate, and Triton X100. An effective amount of said agents may be found between 0.01M:0.01M and 0.4M:0.4M.

[0103] Alternative denaturing agents and surfactants, according to the general knowledge of the person skilled in the art, may be used in the absorbent body in order to guarantee its safety with regard to human and animal health, if for example the device were to be used on living organisms, for example for medical analysis purposes. Under these conditions, the absorbent body may for example be directly applied to the oral cavity of an individual in order to collect cells for genetic analysis purposes.

[0104] Whatever its form and nature, the absorbent body thus comprising surfactant and denaturing agents has the property of trapping and then releasing the collected nucleic acids, thus making them directly available, without delay, for molecular biology applications without it being necessary to carry out a conventional extraction step in the laboratory.

[0105] According to a preferred embodiment of the present invention, said end presenting said absorbent body of the device is breakable or ejectable.

[0106] According to a preferred embodiment of the present invention, said device comprises a hermetic case into which the entire device is inserted, or a part of the rod as well as the end of the rod with the absorbent body, excluding the gripping and handling area, or only the absorbent body. Preferably, the hermetic case comprises a unique identifier, for example by barcode, RFID, or NFC. Preferably the unique identifier according to the invention is a barcode.

[0107] According to a variant of the invention illustrated by the figure 3 , this device may also be coupled to a second collection device consisting of a rod, preferably approximately 5 to 10 cm long, the end of which is preferably breakable or ejectable, having an absorbent body allowing the collection of biological material. This second collection device may, for example, allow the conservation and subsequent extraction of nucleic acids for the purposes of analysis by molecular biology techniques, in particular by PCR. In this configuration, the two devices are associated but each inserted into a separate airtight case, preferably identifiable in particular by a barcode, RFID or NFC.

[0108] The absorbent body of this second collection device may be of the same size and / or composition as or different from the absorbent body of the first device. Preferably, said absorbent body of the second device is made of synthetic fibers flocked onto the stem or cotton fibers wrapped around the stem, with a surface area of ​​between 1mm2 and 2 cm2.

[0109] Advantageously according to the invention, the collection device and in particular its absorbent body, undergoes, at the end of manufacture and before use, a treatment by any type of dry sterilization system, in particular gamma irradiation, ethylene oxide and plasma. This sterilization treatment makes it possible to prevent contamination of the collection device before use by biological material other than that which will be collected. This allows in particular the collection device to meet the most demanding international recommendations regarding the prevention of any contamination by nucleic acids not originating from the biological trace collected.

[0110] The particularly preferred device according to the invention has a solid and dry absorbent body composed of synthetic fibers flocked on the stem, with a surface area of ​​between about 1 and about 3.14 mm2, breakable, on a straight stem 5 to 10 cm long, associated with an identifiable hermetic case. The absorbent body comprises 2 micromoles of uric acid, 8 micromoles of Tris, 0.5 micromoles of EDTA and 1 mg of SDS per cm2. This particularly preferred device is used without prior humidification of the biological material or the absorbent body before collection.

[0111] Preferably according to the invention, the container consists of the well(s) of a plate or strip, for example 8-well, 96-well or 384-well, and is ready to use for PCR. Thus, preferably, the first container is pre-filled with the nucleic acid PCR amplification reaction mixture. Advantageously, said container also comprises the amplification reaction controls, pre-filled in the container and already added to the ready-to-use amplification mixture. The second container is preferably pre-filled with the sequencing reaction mixture of the nucleic acid PCR amplification products. Advantageously, said container may also be pre-filled, in addition to the sequencing reaction mixture, with a set of reference DNA fragments, commonly called an allelic ladder, allowing the identification of the amplification products.

[0112] By container is meant a container for the absorbent body in order to carry out the contacting of said absorbent body with the nucleic acid PCR amplification reaction mixture and the PCR as well as the contacting of the amplification products with the sequencing reaction mixture. For example, according to the present invention, it is intended to denote a container with a capacity of 10 to 350 microliters, or a combination of containers with a capacity of 10 to 200 microliters such as a 96-well PCR microplate, or a 384-well PCR microplate. Preferably, the container is a 96-well PCR microplate of 100 or 200 microliters.

[0113] The time taken to obtain results by using said biological material collection device with the mobile laboratory according to the invention is thus considerably reduced while retaining the advantages of producing analyses in series of several dozen samples. Thus, they make it possible to obtain results on a much larger volume of samples and in much shorter times than the conventional analytical process: A maximum of 2.5 hours to obtain the results for the first 21 samples and 25 minutes for the following 21. In addition, the use of said biological material collection device with the mobile laboratory eliminates the extraction step which is a source of DNA contamination, which is one of the crucial problems to avoid in the context of DNA analysis, particularly at crime scenes.

[0114] Other advantages and characteristics of the invention will appear more clearly on reading the description of a preferred embodiment which follows, with reference to the appended drawings, produced as indicative and non-limiting examples and in which: there figure 1 is a sectional view from above of the mobile laboratory figure 2 is a side view of the entrance to the laboratory. figure 3 is a front view of the instrument airlock side. figure 4 is a rear front view of the mobile laboratory vehicle (trunk (9)) The figure 5 is a view of the maximum configuration where the mobile laboratory has an additional space for sampling and storage of samples (11), and an additional space serving as an interpretation area and a meeting area (11) these additional spaces being mobile and inflatable relative to the mobile laboratory.

[0115] The mobile laboratory is divided into two distinct areas: pre-PCR and post-PCR, physically separated and with separate personal access. The principle of forward movement of personnel and samples is respected: samples are introduced into the laboratory through the serving hatch with alternating door opening communicating between the trunk (9) of the vehicle and the pre-PCR laboratory area. Samples pass from pre-PCR to post-PCR via another serving hatch. Sample sampling / traceability can be achieved by deploying a modular extension from the rear of the vehicle. Data processing and results communication can be achieved by deploying a lateral modular extension providing access to the entrance airlocks of the laboratory areas.The vehicle is equipped with a secure, mobile 4G computer network to transmit analysis results directly to European genetic databases. Technicians communicate with each other via an intercom positioned at each sample serving hatch. The same applies between the dressing room and the laboratory area. HSIE regulations are also taken into account (presence of a fire extinguisher, containers for managing DASRI and a first aid kit in each area).

[0116] Based on the information provided above, it is therefore possible to carry out DNA analyses as close as possible to the crime scene using the same materials and protocols as those used in a conventional laboratory with an equivalent degree of reliability and quality, and very quickly.

Claims

1. Mobile laboratory (1) formed by a transportable mobile container defining a base room, the interior space of said room being isolated from the outside by means of at least one closure system (8), said room comprising: ∘ at least two separate spaces i. The first "pre-PCR" space (2) suitable for carrying out a pre-amplification step and comprising: • A nucleic acid PCR amplification reaction mixture • A receptacle ii. a second "post-PCR" space (3) suitable for carrying out a step of nucleic acid PCR amplification, sequencing and genotyping comprising: • A PCR analysis device (22) • A sequencer (20) • A sequencing reaction mixture • A receptacle ∘ an alternately opening airlock system (4a) between the outside and the pre-PCR area (2), of a size suitable for the passage of a sample and not a human being, said alternately opening airlock system (4) enabling the transmission of samples between the outside and the pre-PCR space (2); ∘ a closure system (4b) between the two spaces (2) and (3), said closure system (4b) sealing the two spaces (2) and (3) off from each other and enabling the transmission of samples between the two spaces (2) and (3); ∘ an autonomous power inverter associated with batteries; said mobile laboratory being suitable for genetic analysis and identification of an individual from a biological sample directly at the location where said sample is taken.

2. Mobile laboratory according to claim 1, characterized in that said sequencer (20) is arranged in an anti-vibration transport device.

3. Mobile laboratory according to any one of claims 1 to 2, characterized in that it comprises an air-conditioning device (13), a secure communication link and a computer network.

4. Mobile laboratory according to any one of claims 1 to 3, characterized in that it comprises at least one instrument pass-through (6).

5. Mobile laboratory according to any one of claims 1 to 4, characterized in that it comprises an additional mobile space (11) for sampling and / or sample storage, and / or an interpretation area, and / or a meeting area.

6. Mobile laboratory according to claim 5, characterized in that said additional mobile space is installed in at least one complementary inflatable and / or deployable space.

7. Mobile laboratory according to any one of claims 1 to 6, characterized in that the PCR analysis device (22), the sequencer (20) and the device ensuring an autonomous, continuous and backed-up power supply are mobile and can be removed from the mobile laboratory after each use of said laboratory.

8. Mobile laboratory according to any one of claims 1 to 7, characterized in that it comprises a biological material collection device comprising a rod having, at at least one end, a dry absorbent body comprising surfactants and denaturants.

9. Vehicle characterized in that it comprises, on the inside, a mobile laboratory according to any one of claims 1 to 8.