EP3454986A1 - Mobile genetic analysis laboratory - Google Patents
Mobile genetic analysis laboratoryInfo
- Publication number
- EP3454986A1 EP3454986A1 EP17725653.4A EP17725653A EP3454986A1 EP 3454986 A1 EP3454986 A1 EP 3454986A1 EP 17725653 A EP17725653 A EP 17725653A EP 3454986 A1 EP3454986 A1 EP 3454986A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- pcr
- mobile
- space
- laboratory
- mobile laboratory
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L1/00—Enclosures; Chambers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/14—Process control and prevention of errors
- B01L2200/141—Preventing contamination, tampering
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/18—Transport of container or devices
- B01L2200/185—Long distance transport, e.g. mailing
Definitions
- the present invention relates to a mobile laboratory for the serial analysis of nucleic acids from a biological sample, particularly in the field of forensics.
- a team of specialists takes samples (blood samples, bone marrow, oral cells, degraded or non-degraded human biological tissue) on a crime scene. These samples are then packaged and transported by a 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 (s) is transmitted to the competent authorities who take the appropriate measures.
- the specialized vehicles carrying the samples are at best refrigerated vehicles or at least vehicles equipped with a cooler, no diagnosis can be made within the same vehicle.
- constraints encountered for the implementation of such a mobile laboratory are the environmental constraints related to the risks of DNA contamination as well as the traditional equipment necessary for the amplification of nucleic acids and in particular the material necessary for sequencing. that are not designed to be moved from their original installation site. These constraints are fundamentally different from those of a laboratory that would be designed to identify pathogens in a CBRN framework (nuclear, radiological, biological and chemical weapons), where the operator would be protected from the outside environment and the dangerousness of the sample, whereas in the case of the mobile laboratory according to the invention, it is sought protection of the sample against contamination by the operator and the external environment / analysis.
- CBRN framework nuclear, radiological, biological and chemical weapons
- 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 the identification of individuals by genetic analysis from a sample, under analysis conditions at least as reliable as in a conventional laboratory so as not to be able to question the reliability of the results.
- This problem must be solved while allowing easy implementation by personnel traditionally performing these operations in a conventional genetic analysis laboratory, without the need for additional training, and at an acceptable cost, particularly for occasional use by the forces police or army where the cost is borne by the community.
- this mobile laboratory can be transported as close to any type of crime scene regardless of its geographical location.
- the Applicant has solved this problem by developing a mobile laboratory of simple design, including the genetic analysis equipment used routinely in a traditional laboratory and adapted for the mobile laboratory when it is necessary to move them, not requiring additional staff training, using the same equipment and the same protocols and quickly operational on the crime scene, and likely to be easily transported by road, but also by sea, rail or air.
- Said mobile laboratory has a configuration which makes it possible to respect the principle of the "step forward" of the treatment of a sample, 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 the PCR gene amplification are performed;
- Another object of the invention is to provide a mobile laboratory in a vehicle, moving autonomously on the road.
- the mobile laboratory according to the invention allows the serial analysis of nucleic acids outside the conventional genetics laboratory thus allowing to have a capacity of mass analysis closer to the sampling scene.
- the delay between the installation of the on-site laboratory, 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 generates at least twenty-one additional results every twenty to twenty-five minutes. The delays in obtaining the results are thus considerably reduced compared to the time required for routing and analysis in a conventional laboratory while retaining the advantages of a series of analysis of several tens of samples.
- This mobile laboratory is designed to respond quickly to crime scenes, for example in the context of a terrorist attack, and to carry out a formal identification of victims or suspects of the offense directly in the area concerned, in order to inform in real time the competent authorities to quickly take the necessary decisions as to the measures to be put in place (confirmation or disruption of the identity of the victim (s) to the families, warrant of arrest against a suspect, call to witness ... ).
- a biological material collection tool comprising a rod having at least one end a dry absorbent body comprising surfactants and denaturants, eliminates the step of DNA extraction and thus reduces the cost of labor, equipment and laboratory consumables necessary for genetic analysis, while reducing the risk of contamination of the sample.
- a first object of the present invention relates to a mobile laboratory (1) formed of a transportable mobile container defining a base room, the interior space of said room being isolated from the outside via at least one control system. closure (8), said room comprising:
- pre-PCR adapted to carry out a pre-amplification step and comprising:
- post -PCR a second space (3) called "post -PCR" adapted to carry out a nucleic acid amplification step by PCR, sequencing and genotyping comprising:
- a closure system (4b) between the two spaces (2) and (3) said closure system (4b) making the two spaces (2) and (3) sealed from one another and allowing the transmission of samples between the two spaces (2) and (3); a device guaranteeing an autonomous, continuous and rescued power supply.
- Said mobile laboratory being adapted to genetic analysis and the identification of an individual from a biological sample directly at the sampling site of said sample.
- closure system is meant according to the present invention a door, preferably sliding, and allowing sealing between the two spaces it separates once closed.
- the closure system (8) separates the outside of the mobile laboratory from the post-PCR space.
- the user of the mobile laboratory according to the invention then enters the said laboratory by the closure system (8) in the post-PCR space, closes the closure system (8), and then returns to the pre-PCR space via the closure system (4). It then closes the closure system (4) to seal between the two spaces.
- the invention may comprise two closure systems (8), one between the outside of the mobile laboratory (1) and the pre-PCR space (2), and the other closure system between outside the mobile laboratory (1) and the post-PCR space (3).
- This configuration is shown in Figures 1 and 2.
- the invention may include an airlock, which is accessed by the closure system (8), which allows a user of the mobile laboratory to access from the outside inside the mobile laboratory, 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 can not be opened together and both can only open when the closure system (8) between the outside and the airlock is closed, in order to prevent any contamination.
- said closure system (8) is an airlock with alternating opening.
- the mobile laboratory according to the invention comprises two airlock chambers, one between the outside of the mobile laboratory (1) and the pre-PCR space (2), and the other between the outside of the laboratory. mobile (1) and post-PCR space (3). Said chambers constitute an area of clothing between the outside and the inside of each analytical zone in which the manipulator can remove his clothing and put on an outfit dedicated to the activities of the laboratory.
- the closure system (4) between the pre-PCR space and the post-PCR space is an alternating opening lock system (4) between the two spaces (2) and (3) of suitable size for the passage of a sample, said alternating opening lock system (4) making the two spaces (2) and (3) watertight from one another and allowing the sample communication between the two spaces (2 ) and (3).
- said airlock does not allow the passage of a human being.
- pre-PCR space (2) is not sized to allow the passage of a human being. Indeed the user of the mobile laboratory enters it inside said laboratory by the closure system (8). This system makes it possible to respect the principle of "walking forward" of the samples (and of the user of the mobile laboratory) and thus to fight at best against the contamination of the samples.
- the nucleic acid PCR amplification reaction mixture as present in the mobile laboratory pre-PCR space refers to a particular combination of reagents that are necessary to perform an amplification reaction. PCR.
- the amplification reaction mixture used in each case and the conditions applied to perform the amplification are well known to those skilled in the art.
- 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 needed to perform amplification.
- the amplification reaction medium can typically amplify STR (Short Tandem Repeat) DNA fragments such as in the kits Identifiler plus and Globalfiler (Lifetechnologies) 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.
- STR Short Tandem Repeat
- the pre-PCR space also comprises at least one bench including one covering a refrigerated chamber of + 4 ° C to -20 ° C (21), a laminar flow hood, at least one container and the small ones.
- basic materials vortex, centrifuge, plate sealer, micropipettes).
- the container consists of the well (s) of a plate or bar, e.g. 8-well, 96-well or 384-well and is ready for use for PCR.
- the container is pre-filled with the nucleic acid PCR amplification reaction mixture.
- said container also comprises the amplification reaction controls, pre-filled in the container and already added to the ready-to-use amplification mixture.
- container is meant a container for the absorbent body for contacting the absorbent body of a biological material collection device with the nucleic acid PCR amplification reaction mixture and the PCR and the contacting the amplification products with the sequencing reaction mixture.
- 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 384 wells.
- the container is a 96-well PCR microplate of 100 or 200 microliters.
- the pre-PCR space is a glove box.
- glove box is meant according to the present invention a sealed enclosure that allows manipulation in a particular atmosphere. Gloves up to the shoulders, attached to one of the walls, allow access to the interior without confinement ceases. The user places his hands in the gloves and then sees his manipulations through the transparent wall.
- the Post-PCR space also comprises at least one bench, one of which covers a refrigerated chamber of + 4 ° C to -20 ° C (21) at least one container and small basic materials (vortex, centrifuge , micropipettes, plate sealer).
- the container is advantageously as described above.
- 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 that are necessary to effect sequencing of amplified DNA fragments.
- the sequencing reaction mixture used in each case and the conditions applied to perform the sequencing by the electrophoresis technique are well known to those skilled in the art.
- the mixture of the sequencing reaction may comprise formamide as well as a set of known molecular weight DNA fragments commonly referred to as 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 scale by those skilled in the art.
- the container is preferably pre-filled with the sequencing reaction mixture of nucleic acid PCR amplification products.
- said container may also be pre-filled, in addition to the mixture of the sequencing reaction, with a set of reference DNA fragments, commonly referred to as an allelic scale, enabling the identification of the amplification products.
- a P.C.R. Polymerase Chain Reaction
- a P.C.R. Polymerase Chain Reaction
- This PCR analysis device (22) is advantageously a thermal cycler. It can be any commercially available thermal cycler for the equipment of a traditional genetic analysis laboratory. Preferably, it has an electrical plug meeting international standards such as a type A and B type C or C, E and F. Thus it may be for example a thermocycler GeneAmp PCR models System 9700, or Veriti marketed by Thermofisher.
- a box to facilitate the transport of said PCR analysis device (22) can be used.
- thermocycler Thermofisher Veriti model
- Genotyping PCR products is then performed using a sequencer analyzer device (20), called “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 the equipment of a traditional genetic analysis laboratory. Preferably, it has a standard electrical outlet. Thus it may be for example a sequencer (20) model ABI 3500 XL marketed by the company Thermofisher.
- the sequencer (20) is generally an imposing instrument that can weigh a hundred kilos and sedentary that uses sensitive optical systems may become unstable over time. It is therefore a critical instrument to transport.
- the optics of the sequencer (20) may be subject to a greater fixation, namely that the optics fixing system of the sequencer (20) has been tighter 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.
- sequencer There are many models of sequencer and it is not possible to define a standard tightening that could be applied to any sequencer of the market, nor those that could be commercialized in the future. Indeed each sequencer has its own otic system and its own original clamping force.
- a calibration for example by testing the original clamp and then different tightening of the optical system of the sequencer so as to whether it is transported, or subject to conditions similar to a transport (for example by placing sequencer on a plate subjected to controlled movements of back and forth, circular, vibration and oscillations at a magnitude between 500 microns up to 1cm, at a frequency of 5 to 50 Hz), without the optical systems are disrupted.
- Such a calibration on the capillary electrophoresis systems may for example be carried out using known molecular weight DNA fragments labeled with fluorescent molecules in order to be excited by the laser using the optical system of the sequencer, then to be detected. by the camera.
- the optics system is disordered then the DNA fragments will not be detected by the camera or at molecular weights different from those expected.
- the tightening of the optical system can be conventionally carried out using a dynamometric tool such as a screwdriver.
- all the threads of the optical system of the sequencer can be treated with a weak braking product of the LOCTITE® 221 TM type which is a polymer designed for braking and sealing the threaded connections. Its function is to prevent loosening or leakage due to shock and vibration.
- 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 with respect to said sequencer (20) and lined with a vibration-damping viscoelastic material such as a damping polymer foam.
- 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 35 kg / m3 and 105kg / m3.
- Another anti-vibration transport device may comprise VIBCABLE type cable dampers marketed by PAULSTRA.
- a calibration procedure is performed. This is the same procedure as that performed when installing the sequencer (20) when initially delivered to a traditional laboratory.
- said mobile laboratory may comprise several thermocyclers and / or sequencers in order to increase the number of samples that can be identified over a given period of time.
- the computer cockpit has also been changed from the standard version: a laptop replaces the sedentary computer.
- the mobile laboratory comprises a lifting table for transporting the transport boxes of the thermal cycler (22) and / or the sequencer (20) inside and outside the mobile laboratory.
- the sequencer (20) and the thermal cycler (22) are inserted and removed from the mobile laboratory via a pass alternately opening instrument (6) consisting of a "rolling shutter” or a "pivoting wall” accessible from outside the vehicle and a shutter accessible from inside the laboratory in the post-analytic zone; PCR, to protect the laboratory area from the external environment and the entry of individuals into this area.
- the mobile laboratory according to the invention comprises at least one instrument-holder (6) and, more preferably, a instrument-holder (6) for each device, consisting of a shutter or a pivoting wall giving access from the each external to an airlock alternately opening with the interior of said laboratory, or advantageously a shutter (6s) or a pivoting wall for the passage of the sequencer and giving access to a sequencer lock, and a shutter (6t) or a pivoting wall for the passage of the thermal cycler, and giving access to a thermocycler airlock;
- the base of the instrument (6s) of the sequencer (20) can be raised by a mechanical action engaged from inside the post-PCR zone in order to constitute a working plane at standard height (85-90cm from vehicle floor).
- the storage area of the cover of the transport case of the sequencer (20) is located under the base of the instrument-holder (6s) once it is in the up position.
- An equivalent instrumentation system (6t) is developed for the thermal cycler (22) without the need for a lifting pedestal.
- An equivalent roller shutter system for the generator set is developed.
- the instrument-holders have hooks for securing the instrument transport crates to ensure their maintenance when the vehicle is moving.
- the critical materials can be inserted and removed from the laboratory without having to enter the analytical zone thus preserving these areas from all soils and contaminations from the external environment.
- 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 passage.
- the mobile laboratory according to the invention also however comprises a generator (7), and a 12v / 220V converter (or 110V) electrical switchboard according to the country of use of the mobile laboratory.
- the autonomous inverter delivers a voltage with a frequency that is fixed or adjustable by the user. It does not always need a power grid to work; for example a travel converter that plugs into the cigarette lighter socket of a car uses the 12 V DC of the vehicle to generate 120 or 230 V, alternating in 50 or 60 Hz;
- the device ensuring an autonomous, continuous and rescued power supply is stored in the trunk (9) of the vehicle.
- the location (5) corresponds to the electrical panel of the laboratory area (12v / 220v converter) and the location of the inverter and associated batteries.
- the device ensures uninterrupted power supply throughout the PCR amplification and sequencing process.
- the laboratory according to the invention can thus operate completely autonomously on crime scenes.
- 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 conditions of ambient temperature. Indeed the environmental conditions having an impact on the electrophoresis conditions during the genotyping with the sequencer, it is important to be able to control the ambient temperature to attest that the analyzes are carried out in a temperature range answering the specifications of the sequencer.
- the laboratory according to the invention has an independent air conditioning device (13) for each pre-PCR and post-PCR space.
- said device also makes it possible to control the conditions ambient hygrometry and limit the contamination of spaces by dust.
- said mobile laboratory according to the present invention also comprises a continuous temperature recorder for tracing the conditions of use of the instruments.
- a refrigerated chamber + 4 ° C / -20 ° C is also present in each zone. Their temperature is also plotted.
- the mobile laboratory according to the invention is characterized in that it comprises a computing device, such as at least one portable computer, an internal computer network as well as a secure telematic link.
- a computing device such as at least one portable computer
- an internal computer network as well as a secure telematic link.
- Said internal computer network makes it possible to exchange data in the internal perimeter of the mobile laboratory.
- Said telematic link allows for all over the world to transmit the information and in particular the genetic profiles thus determined for example to an expert or in national or international databases.
- the telematic link also transmits video via cameras installed in each of the technical zones, which makes it possible to record all the technical operations performed but also to view all these technical operations in real time from outside the mobile laboratory and to distance.
- Such a telematic link is for example a wireless internet connection means such as a 4G key, or a satellite link associated with an encryption or encryption means.
- the mobile laboratory according to the present invention comprises an air conditioning device (13), a secure telematic link, and a local computer network.
- closure system is meant to designate according to the present invention a door, preferably sliding, and allowing sealing between the two spaces that it separates once closed.
- the closure system (8) separates the outside of the mobile laboratory from the post-PCR space.
- the user of the mobile laboratory according to the invention then enters the said laboratory by the closure system (8) in the post-PCR space, closes the closure system (8), and then returns to the pre-PCR space via the closure system (4). It then closes the closure system (4) to seal between the two spaces.
- the user of the mobile laboratory according to the invention can enter inside said laboratory by the closure system (8) in the pre-PCR space and then access via the system of closure (4) at the post-PCR space.
- the invention may comprise two closure systems (8), one between the outside of the mobile laboratory (1) and the pre-PCR space (2), and the other closure system between outside the mobile laboratory (1) and the post-PCR space (3).
- This configuration is shown in Figures 1 and 2.
- said closure system (8) is an airlock with alternating opening.
- the mobile laboratory according to the invention comprises two airlock chambers, one between the outside of the mobile laboratory (1) and the pre-PCR space (2), and the other closure system between the outside the mobile laboratory (1) and the post-PCR space (3).
- Said chambers constitute an area of clothing between the outside and the inside of each analytical zone in which the manipulator can remove his clothing and put on an outfit dedicated to the activities of the laboratory.
- the closure system (4) between the pre-PCR space and the post-PCR space is an alternating opening lock system (4) between the two spaces (2) and (3) of suitable size for the passage of a sample, said alternating opening lock system (4) making the two spaces (2) and (3) watertight from one another and allowing the transmission of samples between the two spaces (2 ) and (3).
- said airlock does not allow the passage of a human being.
- each airlock or closure system in the mobile laboratory is equipped with an intercom for communicating with the outside or the other spaces of the mobile laboratory.
- the mobile laboratory according to the invention is characterized in that it further comprises an additional space (11) Sampling used for biological sampling and may include, for example, sampling tables such as autopsy tables.
- 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.
- 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 link telematics.
- the mobile laboratory according to the invention is characterized in that it furthermore comprises a meeting zone making it possible to plan the analyzes and to communicate the results obtained.
- the mobile laboratory according to the invention is characterized in that it comprises a mobile additional space (11) for sampling and / or sampling storage, and / or an interpretation zone. , and / or a meeting area.
- the mobile laboratory comprises at least one complementary space (11) mountable on the spot at the place of the infringement and forming at least one annex in which said additional sampling space can be installed, said sample storage space, said interpretation area or said meeting area.
- the total area of the mobile laboratory and its annexes is approximately 40 m2 which is enough space to work in good conditions.
- the mobile laboratory according to the invention is characterized in that said complementary spaces (11) are mobile.
- said complementary spaces are inflatable and / or deployable. In particular, they may be deployable walls from the mobile laboratory.
- the mobile laboratory according to the invention is characterized in that the small basic materials, the reagents and consumables necessary for the analyzes, the PCR analysis device (22), the sequencer (20) and the device guaranteeing autonomous, continuous and emergency power supply 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 these instruments to the mobile laboratory only when it is necessary, and without it being necessary. necessary to possess expensive equipment, in particular a PCR analysis device (22) and a sequencer (20) for each of the mobile and traditional laboratories.
- the laboratory is arranged inside a vehicle.
- vehicle By vehicle is meant to designate according to the present invention a motorized mobile vehicle, typically a van or a bus.
- a motorized mobile vehicle typically a van or a bus.
- said motorized vehicle behaves with a driver's license for passenger vehicles (Permit B in France). This allows in case of emergency intervention to expand the number of people who can bring said mobile laboratory to an area of intervention.
- the mobile laboratory is arranged directly inside said mobile vehicle.
- said vehicle has at least one sound and light horn (14) allowing its signaling to other users when en route to the scene of offense and facilitate the transport of the vehicle as quickly and safely as possible .
- a storage area for said lift table is dedicated in the trunk (9) of the vehicle of the mobile laboratory.
- the mobile laboratory according to the invention has external electrical outlets (15) for connection to the electrical network, especially if a generator is not used, and electrical outlets internal to the analytical area, as well as lighting in particular if said laboratory does not include windows.
- the mobile laboratory according to the invention comprises a system for stabilization and horizontal leveling of the vehicle at a standstill which could be useful depending on the configuration of the terrain on the projection site.
- the mobile laboratory is movable by any means, road, air, rail or sea.
- the mobile laboratory can be moved by air by being loaded on board a military aircraft, for example of the C-160 TRANSALL type.
- the mobile laboratory according to the invention is characterized in that it comprises a device for collecting biological material comprising a rod having at at least one end a dry absorbent body comprising surfactants and denaturants.
- a dry absorbent body comprising surfactants and denaturants.
- said absorbent body has a surface area of between about 1 and about 3.14 mm 2
- biological material is intended to mean, according to the present invention, any material derived from a living being, which may be in the form of molecules, organelles, fluids, cell fragments or cells, and present on a biological or non-biological support organic.
- 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 trace of contact, blood, bone marrow, buccal cells, tissue degraded or undegraded human biology 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 derived from nucleic acid synthesis; and mixtures of nucleic acids synthesized chemically or biochemically.
- the biological material according to the present invention is in particular a trace of contact, blood, bone marrow, buccal cells, degraded or non-degraded human biological tissue.
- nucleic acids is meant according to the present invention DNA and RNA.
- DNA and RNA are meant according to the present invention the DNA.
- absorbent body is meant according to the present invention a dry solid support composed of an absorbent material, natural or synthetic.
- said absorbent body is selected from the group consisting of cellulose, nitrocellulose, cotton, flocked synthetic fibers, e.g. flocked nylon, hydrophilic polymers, fiberglass, polytetrafluoroethylene, polyester, nylon, rayon, cotton and a porous ceramic. More preferably, said absorbent body is selected from cellulose, and flocked synthetic fibers.
- said absorbent body is composed of flocked synthetic fibers, preferably flocked nylon fibers, preferably directly on the rod.
- the fibers are glued to the rod.
- the fibers are between 1 and 1.5 mm in length and between 0.01 and 0.5 mm in diameter. This configuration makes it possible to obtain the best results in terms of collecting biological material and releasing the nucleic acids of said biological material, combined with simplicity of manufacture and use.
- the absorbent body again has a major economic interest because this characteristic makes it possible to carry out the amplification of nucleic acids in a reduced reaction volume of 50% compared to the supplier's recommendations and therefore considerably reduce the cost of analyzes of a large number of samples.
- the tests carried out with the gene amplification kits Identifiler plus and Globalfiler enabled, for example, to demonstrate the effectiveness of the device for a 25 ⁇ PCR reaction volume as recommended by the supplier, but also in a reduced reaction volume. in half that is 12,5 ⁇ 1. It has thus been demonstrated that such a small surface of the absorbent body makes it possible to collect a sufficient quantity of nucleic acids to obtain a genetic profile.
- Said absorbent body is pre-treated with denaturants and surfactants during its design thus allowing in situ lysis of the cells. It is therefore not necessary to apply on this absorbent body a specific humidification buffer for the lysis of the cells upstream of the collection.
- surfactants and denaturants agents for cell lysis, denaturation of proteins and extraction of nucleic acids.
- the surfactants and denaturants may be chosen from a list comprising uric acid or urate salt, tris, SDS guanidine salt, preferably guanidine thiocyanate, Triton XI 00, Tween 20, CHAPS, CHAPS-O, octyl -glucoside, octyl thioglucoside and combinations thereof.
- the surfactants and denaturants are a combination of uric acid or urate salt, tris, and SDS.
- the Absorbent body further comprises EDTA.
- the absorbent body comprises a mixture per cm 2 of 2 micromols of uric acid, 8 micromols of Tris, and 1 mg of SDS.
- the absorbent body further comprises 0.5 micromols of EDTA.
- the surfactants and denaturants may be a combination of guanidine salt, preferably guanidine thiocyanate, and Triton XI 00.
- An effective amount of said agents can be found between 0.01M: 0.01M and 0.4M: 0.4M.
- the absorbent body may be used in the absorbent body to ensure its safety with respect to human and animal health, if for example the device was to be used on living organisms, for example for medical analysis purposes. Under these conditions, the absorbent body may for example be directly applied in the oral cavity of an individual to collect cells for genetic analysis.
- the absorbent body thus comprising surfactants and denaturants has the property of trapping and releasing the collected nucleic acids thus made directly available, without delay, for molecular biology applications without it being necessary. to perform a conventional extraction step in the laboratory.
- said end having said absorbent body of the device is breakable or ejectable.
- said device comprises a hermetic case in which is inserted the entire device, or a part of the stem and the end of the rod with the absorbent body, excluding the the area of grip and manipulation, or only the absorbent body.
- the hermetic case comprises a unique identifier, for example by barcode, RFID, or NFC.
- the unique identifier according to the invention is a barcode.
- this device may also be coupled to a second collection device consisting of a rod, preferably of a length of approximately 5 to 10 cm, preferably of which end is breakable or ejectable, has an absorbent body allowing the collection of biological material.
- This second collection device can, for example, enable the preservation and subsequent extraction of nucleic acids for analysis by molecular biology techniques including PCR.
- the two devices are associated but each inserted into a separate sealed case, preferably identifiable in particular by a barcode, RFID or NFC code.
- 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.
- said absorbent body of the second device is made of synthetic fibers flocked on the rod or cotton fibers wound around the rod, with an area of between 1 mm 2 and 2 cm 2.
- 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 and plasma.
- This sterilization treatment prevents contamination of the collection device before use by biological material other than that which will be taken. This allows in particular the collection device to meet the most demanding international recommendations in terms of prevention of any contamination by nucleic acids that do not come from the biological trace taken.
- the particularly preferred device according to the invention has a solid and dry absorbent body composed of synthetic fibers flocked to the stem, with an area of between about 1 and about 3.14 mm 2, breakable, on a rectilinear stem of 5 to 10 cm length, associated with an identifiable hermetic case.
- the absorbent body comprises 2 micromols of uric acid, 8 micromols of Tris, 0.5 micromols of EDTA and 1 mg of SDS per cm 2. This particularly preferred device is used without prior wetting of the biological material or the absorbent body before collection.
- the container consists of the well or wells of a plate or a strip, for example of 8 wells, 96 wells or 384 wells and is ready for use for the PCR.
- the first container is pre-filled with the nucleic acid PCR amplification reaction mixture.
- 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 nucleic acid PCR amplification products.
- said container may also be pre-filled, in addition to the mixture of the sequencing reaction, with a set of reference DNA fragments, commonly referred to as an allelic scale, enabling the identification of the amplification products.
- container is meant a container for the absorbent body to effect contact of said absorbent body with the nucleic acid PCR amplification reaction mixture and the PCR and the contacting of the amplification products with the sequencing reaction mixture.
- 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 384 wells.
- the container is a 96-well PCR microplate of 100 or 200 microliters.
- Figure 1 is a sectional view from above of the mobile laboratory
- FIG. 2 is a side view of the laboratory entrance
- FIG. 3 is a front view of the instrument
- FIG. 4 is a rear view of the vehicle of the mobile laboratory (trunk (9))
- FIG. 5 is a view of the maximum configuration where the mobile laboratory
- sampling 11
- additional space serving as interpretation area and a meeting area (11) these additional spaces being mobile and inflatable relative to the mobile laboratory.
- the mobile laboratory is compartmentalized into two distinct spaces: pre-PCR and post-PCR physically separated and separate personal access.
- the principle of forward movement of personnel and samples is respected: the samples are introduced into the laboratory by the alternating opening door opening communicating between the trunk (9) of the vehicle and the pre-PCR laboratory area.
- the samples pass from the pre-PCR to the post-PCR via another drier.
- Sampling / traceability of samples can be achieved by deploying a modular extension from the rear of the vehicle.
- the exploitation of the data and the communication of the results can be carried out by the deployment of a lateral modular extension giving access to the entrance locks of the laboratory zones.
- a 4G network secure and mobile computer network equips the vehicle to transmit test results directly into European genetic databases.
- the technicians communicate with each other via an intercom positioned at each sample drip tray. Ditto between the airlock and the laboratory zone.
- HSIE rules are also taken into account (presence of a fire extinguisher, containers for the management of DASRI and a pharmacy kit in
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Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR1670215A FR3050945B1 (en) | 2016-05-09 | 2016-05-09 | MOBILE LABORATORY FOR GENETIC ANALYSIS |
PCT/FR2017/000083 WO2017194849A1 (en) | 2016-05-09 | 2017-05-09 | Mobile genetic analysis laboratory |
Publications (1)
Publication Number | Publication Date |
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EP3454986A1 true EP3454986A1 (en) | 2019-03-20 |
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ID=57796728
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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EP17725653.4A Pending EP3454986A1 (en) | 2016-05-09 | 2017-05-09 | Mobile genetic analysis laboratory |
Country Status (3)
Country | Link |
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EP (1) | EP3454986A1 (en) |
FR (1) | FR3050945B1 (en) |
WO (1) | WO2017194849A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
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KR20230112693A (en) * | 2020-12-28 | 2023-07-27 | 주식회사 씨젠 | Mobile Diagnostic Structure |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
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DE102005006756A1 (en) * | 2005-02-15 | 2006-08-17 | Robert Bosch Gmbh | Imaging system |
FR2899918B1 (en) * | 2006-04-14 | 2010-10-01 | Vladimir Grcevic | MOBILE LABORATORY FOR THE ANALYSIS OF PATHOGENIC AGENTS |
-
2016
- 2016-05-09 FR FR1670215A patent/FR3050945B1/en active Active
-
2017
- 2017-05-09 WO PCT/FR2017/000083 patent/WO2017194849A1/en unknown
- 2017-05-09 EP EP17725653.4A patent/EP3454986A1/en active Pending
Also Published As
Publication number | Publication date |
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FR3050945A1 (en) | 2017-11-10 |
WO2017194849A1 (en) | 2017-11-16 |
FR3050945B1 (en) | 2019-01-25 |
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