Test system, detection device, test method, and test preparation device

Abstract

The present invention relates to a test system, a detection device for simultaneously testing samples contained in different test vessels, and a test vessel assembly comprising a single lysis chamber and a dosing assembly selectively fluidly connected to the lysis chamber and comprising a plurality of dosing compartments, the invention further relates to a method for dosing and simultaneously transferring the same portion of the lysed sample from the lysing chamber into the test vial.

Description

[0001] The present invention relates to a test system, a test device, a test method and a test preparation device for detecting a target analyte, in particular a target nucleic acid such as DNA or RNA, by isothermal nucleic acid amplification and fluorescence.

[0002] Nucleic acid amplification techniques are used to amplify the amount of target nucleic acid in a sample in order to detect such target nucleic acid in the sample. A known nucleic acid amplification technique is the Polymerase Chain Reaction (PCR). Isothermal nucleic acid amplification techniques offer advantages over the Polymerase Chain Reaction (PCR) since they do not require thermal cycling or complex laboratory equipment.

[0003] Known isothermal nucleic acid amplification techniques are in particular Recombinase Polymerase Amplification (RPA) and Strand Invasion Based Amplification (SIBA) as well as other methods known to the person skilled in the art.

[0004] Recombinase Polymerase Amplification (RPA) is a known method for amplifying the amount of a target analyte, in particular a nucleic acid such as DNA or RNA, in a sample. For recombinase polymerase amplification, three core enzymes are used: a recombinase, a single-stranded DNA-binding protein (SSB) and a strand displacement polymerase. The recombinase can pair an oligonucleotide primer with a homologous sequence in double-stranded DNA. The SSB binds to the displaced DNA strand and prevents the primer from being displaced. The strand displacement polymerase starts DNA synthesis at the site where the primer binds to the target DNA. Thus, if the target gene sequence is indeed present in the test sample, an exponential DNA amplification reaction can be achieved to amplify a small amount of target nucleic acid to a detectable level within minutes at a temperature between 37 °C and 42 °C.

[0005] The three core RPA enzymes can be supplemented by additional enzymes to provide additional functionality. The addition of exonuclease III allows the use of an exo-probe for real-time fluorescence detection. If a reverse transcriptase that works at 37 °C to 42 °C is added, RNA can be reverse transcribed and cDNA can be amplified in one step.

[0006] By adding reverse transcriptase to the RPA reaction, RNA as well as DNA can be detected without the need for a separate step to generate cDNA. An advantage of RPA is that it is isothermal and thus requires only simple equipment. Although RPA works best at temperatures between 37 °C and 42 °C, it still functions at room temperature.

[0007] To detect the presence of a target nucleic acid in a sample, fluorescence detection techniques can be used. After a light source of a specific wavelength shines on the target nucleic acid, a DNA-binding dye or a fluorophore-binding probe of the nucleic acid will react and enable the emission of a fluorescence signal. The fluorescence signal is an indication of the presence of the target nucleic acid.

[0008] The present invention relates to a rapid and easy-to-handle method for the isothermal amplification of nucleic acids (including DNA and RNA). In particular, the present invention relates to a diagnostic method for rapidly diagnosing at least two infectious agents or at least two different targets in the same infectious agent in a biological sample of interest. The present invention also relates to a handheld and portable diagnostic system for performing the amplification method in a laboratory as well as a non-laboratory environment.

[0009] Nucleic acid amplification techniques (NAATs) (such as molecular real-time PCR assays) are generally very sensitive and specific, but when it comes to the results, PCR still has inherent drawbacks, requiring a highly equipped laboratory and well-trained personnel. Therefore, there is an urgent need for new portable diagnostic solutions that have good specificity and sensitivity and can provide reliable results on-site at the place of testing.

[0010] Regarding nucleic acid-based preparation, cloning, and diagnostic techniques, subsequent Nobel laureate Kary Mullis and the team developed polymerase chain reaction (PCR) in the 1980s as a rapid and reliable method that generally revolutionized molecular biology by amplifying DNA. Simplicity and efficiency (e.g., the current possibility of performing single-molecule / cell PCR) represent significant advantages of the PCR technique.

[0011] However, PCR has certain drawbacks, including the inherent need for multiple thermal cycles and the transfer between different temperatures (repeatedly cycling through two or three temperature-related steps during the amplification process) as well as the use of high temperatures (>90 °C). These drawbacks have led to the development of alternative amplification methods.

[0012] An important class of PCR alternatives are the so-called isothermal amplification methods (for a review, see Zanoli and Spoto, Biosensor (Basel), 2012 3(1): 18-43). A great advantage over PCR is that isothermal nucleic acid amplification methods do not require any thermal cycling at all, but can be carried out at a constant temperature. This makes the amplification process easier to operate and control. In addition, less energy is required than for PCR methods, which inherently require rapid heating and cooling steps. The constant temperature of the isothermal method additionally allows for fully enclosed microstructured devices, where performing isothermal amplification reduces the risk of sample contamination and implies low sample consumption, multiplex DNA analysis, integration, and portable device implementation. Finally, the constant temperature would be highly preferably used for point-of-need and / or portable diagnostic devices, such as those recently developed by the present applicant (DE10 2020 109 744.1, which is incorporated herein by reference).

[0013] Isothermal amplification strategies available to date include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), rolling circle amplification (RCA), multiple displacement amplification (MDA), recombinase polymerase amplification (RPA) (again see: Zanoli and Spoto, 2012, ibid.; for a comprehensive survey of current RPA technology: Li et al., Analyst, 2019, 144, 31, pp. 31 to 67) or strand invasion-based amplification ( - hereinafter - SIBA) (Hoser et al., PLoS ONE 9(11): e112656. doi: 10.1371 / journal.pone.0112656). However, just like PCR isothermal technologies, a major drawback is the tendency to produce non-specific amplification products.

[0014] However, for example, NASBA, RPA, and HDA (as well as certain types of quantitative PCR) rely on the use of target-specific probes to provide higher specificity of the reaction, while LAMP uses additional primers instead of probes and strand displacement polymerase (Notomi et al., 2000, Nucleic Acids Research 28: e63). However, the use of a large number of large primers during at least 30 minutes of incubation has been shown to produce false positive results - a result that must be avoided especially during diagnostic applications for diagnosing infectious diseases. In addition, there is a problem that the LAMP reaction requires a high temperature of 65 °C, which is disadvantageous for diagnostic devices used in a home environment due to regulatory and practical issues. Therefore, if diagnostic nucleic acid amplification results need to be obtained within a short time by a handheld device that must meet certain regulatory requirements for non-laboratory use, LAMP will not be the preferred method.

[0015] Both RPA and SIBA technologies rely on the use of recombinases during the binding and amplification processes. Initially, nucleoprotein complexes composed of oligonucleotide primers and recombinase proteins are formed for RPA and SIB-type nucleic acid amplification strategies, and these complexes facilitate the binding of the primers to the template DNA. Due to their short run times (about 15 minutes), no non-specific amplification has been observed. One advantage is that recombinases can tolerate at least one incorrect base without preventing the strand invasion required to initiate the reaction.

[0016] In parallel with the biochemical progress in nucleic acid amplification, devices including suitable reaction and detection chambers for performing nucleic acid amplification in an optimized manner have also been steadily developed. One major trend is the development of microfluidic devices, and generally, the trend towards miniaturization or "nanosized reaction chambers" (i.e., placing them in a nanoscale form) is of great interest for reducing the sample volume and thus the required reagent amounts, as well as for realizing the advantages of biochemical detection, generation time, faster heat transfer, and the potential for automation and integration, and more for the possibility of multiplexing or multimeric testing. In this context, "multimeric" means multiplexing in the sense of performing parallel reactions in separate (locally distinct) reaction chambers.

[0017] It has been observed that rapid testing for more than one pathogen or target sequence has become very important. For cost reasons, general practitioners usually do not perform dual tests, and general practitioners are usually the first doctors to diagnose patients with symptoms of suspected respiratory diseases. In addition, regarding preventive testing of potentially contaminated surfaces or returning travelers from abroad, it is often observed that the testing is done too late (when the symptoms of the disease become obvious), or given the cost involved in PCR testing, standardized tests that would allow targeted decontamination of surfaces, etc. are not performed regularly.

[0018] WO2021 / 204900A1 and WO2021 / 204901 disclose a test system for detecting a target analyte, different containers containing reagents to be used in the system, and a test device. The detection device of the prior art includes a detection chamber that can receive a container containing a reagent for detecting a target analyte. A light source and a light sensor are arranged inside or adjacent to the detection chamber. To detect the target analyte, the light source irradiates the contents of the detection chamber with light that can cause luminescence in the sample to be tested during and after the sample undergoes recombinase polymerase amplification. The light sensor is arranged and configured to detect the luminescence in the detection chamber in the event that luminescence occurs.

[0019] WO2019 / 060950A1 discloses a diagnostic test system that includes: a diagnostic test assembly and a diagnostic test device for testing a biological or environmental sample. The diagnostic test assembly includes: a sample preparation reservoir for receiving the sample into a sample preparation fluid such that a swab carrying the sample can be used to agitate the preparation fluid and wash the swab; a sample dispensing mechanism for inserting into the sample preparation reservoir; a closure for sealing the sample preparation reservoir; at least one diagnostic test reservoir coupled to the sample preparation reservoir; and at least one seal located between the sample preparation reservoir and the diagnostic test reservoir to prevent fluid movement between the respective reservoirs. The sample dispensing mechanism is operable to break the seal to allow sample fluid to enter the diagnostic test reservoir from the sample preparation reservoir and dispense a predetermined amount of fluid.

[0020] The object of the present invention is to provide a device that allows for easy use and failure-preserving testing of samples for different bioanalytes.

[0021] Therefore, the object of the present invention is to provide a test system, a detection device, and a test method for simultaneously testing different target analytes (especially DNA or RNA target sequences), which can be carried out in a preferably handheld or at least portable diagnostic device under non-laboratory conditions as well as under laboratory conditions, so that point-of-care diagnosis of infections can be carried out even in a home or point-of-care environment, and to provide rapid and reliable diagnostic results for diagnosing multiple potential infections and / or target nucleic acids in the same test sample preferably directly applied to the diagnostic device. The device should be configured in such a way that an easily customizable test for at least two target sequences should be provided, wherein the biochemical reactions that the device allows to be performed can be easily exchanged to adapt to the diagnostic needs of customers interested in the relevant results.

[0022] In particular, the object of the present invention is to facilitate the testing of samples by means of nucleic acid amplification techniques.

[0023] According to a first aspect of the present invention, there is provided a test system, the test system comprising a detection device having a plurality of detection chambers for receiving test containers, the test containers containing a sample to be tested and a chemical mixture, the chemical mixture comprising a target-specific probe and an enzyme, the target-specific probe and the enzyme being capable of causing amplification of nucleic acids in the sample. Different probes are required to test different target nucleic acids, and thus different test containers are required.

[0024] According to a second aspect of the present invention, a test container assembly comprises a single lysis chamber and metering means for dispensing a determined quantity of fluid from the lysis chamber into individual test containers. The lysis chamber contains a liquid lysis fluid which causes lysis of cells in the sample, thereby releasing nucleic acids (DNA or RNA). The lysis fluid may comprise an acid (such as HCl or a weak base) and a surfactant.

[0025] Each test container contains a chemical mixture capable of causing amplification of nucleic acids in the sample. Preferably, the mixture comprises a target-specific probe and an enzyme, in particular a recombinase, a single-stranded DNA binding protein (SSB) and a strand displacement polymerase which causes recombinase polymerase amplification (RPA). The test container also preferably contains exonuclease III, allowing real-time fluorescence detection using an exonuclease probe. The mixture may be provided in the form of dry granules.

[0026] One aspect of an easy-to-use fail-safe test system for simultaneously testing samples for different target nucleic acids is provided by a metering assembly including a lysis chamber and metering means.

[0027] According to a third aspect of the present invention, there is provided a detection device.

[0028] The detection device comprises

[0029] - a plurality of detection chambers, wherein at least one detection chamber body surrounds the detection chamber, each detection chamber being configured to receive a test container,

[0030] - one or more light sources, one or more light sensors,

[0031] - temperature control means,

[0032] - a battery,

[0033] - a data communication interface, and

[0034] - a controller.

[0035] Preferably, the heat capacity of at least one detection chamber corresponds to 5 to 100 times the heat capacity of the fluid sample in the test container. Preferably, the sample volume in the test container is 50 μl / 0.05 gr. The specific heat capacity of the sample corresponds to that of water, i.e., 4184 J / kg*K.

[0036] The test system according to the first aspect preferably includes a detection device having a plurality of detection chambers for receiving test containers containing a sample and a chemical mixture to be tested, the chemical mixture including a target-specific probe and an enzyme, and the target-specific probe and the enzyme being capable of causing amplification of nucleic acids in the sample. The detection device further includes a controller, a memory, and a data communication interface. The memory and the data communication interface are operatively connected to the controller.

[0037] The test system further includes a test container assembly including a plurality of test containers containing a sample and a chemical mixture to be tested, the chemical mixture including a target-specific probe and an enzyme, and the target-specific probe and the enzyme being capable of causing amplification of nucleic acids in the sample. According to a preferred embodiment, one of the containers contains a reference or control assay that will always cause luminescence if the test system is properly processed and the test procedure is performed without fault.

[0038] The test system preferably further includes a smart communication device configured for wireless data communication with the detection device.

[0039] The test system preferably includes a detection device as described below.

[0040] The detection device according to the third aspect preferably includes:

[0041] - a plurality of detection chambers for receiving test containers,

[0042] - at least one light source arranged and configured to irradiate at least one detection chamber to test for luminescence in the sample,

[0043] - at least one light sensor for each detection chamber configured to detect luminescence in the sample contained in the test container placed in the corresponding detection chamber,

[0044] - a controller, and

[0045] - a memory connected to the controller.

[0046] Optional other components of the detection device include a temperature sensor, an inertial measurement unit, a heating device, and a status indicator.

[0047] A light sensor and at least one light source are arranged to allow the contents of the detection chamber to be irradiated by means of the at least one light source, and the luminescence in the respective detection chamber to be detected by means of the respective light sensor assigned to the deflection chamber, while preventing the light emitted by one or more light sources from directly irradiating any light sensor. The light sensor and the at least one light source are preferably arranged transversely with respect to the detection chamber and the test container, respectively, to avoid the negative impact of the particles deposited on the bottom of the test container on the light signal to be measured by the respective light sensor.

[0048] The controller is connected at least indirectly to the light source and the light sensor for controlling the at least one light source and for controlling the reading of the output value of the light sensor. In particular, the controller can be adapted by means of software stored in the memory and by means of drive circuits for the light source and the light sensor to control the illumination of the respective detection chamber by means of the light source and to read out the output signal of the light sensor.

[0049] The light source is preferably a multi-color light source, such as a multi-color LED, which can be controlled by the controller with respect to the color (wavelength band) emitted by the respective light source and with respect to the intensity of the emitted light.

[0050] The memory includes software defining the operation of the controller. The software stored in the memory can include a device operating system allowing the control of the device electronic components by means of the controller.

[0051] The software stored in the memory can also include a script interpreter adapted to interpret script commands stored in the memory. Preferably, the script commands are part of a script defining a test procedure suitable for one or more specific assays contained in the test container. Thus, the memory preferably includes software defining the script interpreter, and the detection device is preferably configured to receive script commands which, when interpreted by the script interpreter and executed by the controller, define the test procedure.

[0052] The memory is also adapted to store at least parameter values corresponding to the output values of the light sensor. Other parameter values can be the output values of a temperature sensor and / or the output values of an inertial measurement unit.

[0053] The detection device preferably includes heating means for controlling the temperature in the detection chamber. The heating means are preferably configured to maintain a predetermined temperature in the detection chamber within a temperature range of + / - 0.5 K with respect to the predetermined temperature, and the heating means are controlled by the controller. The predetermined temperature is preferably a temperature between 40 °C and 45 °C, preferably 42 °C.

[0054] The detection device preferably further includes a metal block, which is thermally connected to the heating device and at least partially surrounds the detection chamber, and the metal block is arranged and configured to provide a uniform heat distribution. The thermal mass of the metal block corresponds to the power of the heating device so as to achieve a suitable temperature gradient during heating and when the temperature is feedback-controlled to remain more or less constant. In other words: the thermal mass of the metal block is selected to allow stable feedback control of the temperature of the detection chamber.

[0055] The detection device preferably further includes a temperature sensor for determining a temperature corresponding to the temperature of at least one of the detection chambers. The temperature sensor allows feedback control of the temperature in the detection chamber.

[0056] The detection preferably further includes a wireless data interface for wirelessly transmitting data to and receiving data from a smart communication device. As will be further disclosed in detail below, data communication with the smart communication device facilitates the use of the detection device in many aspects.

[0057] The detection device preferably further includes a status indicator operatively connected to the controller. The detection device is preferably configured to indicate the current status of the detection device only via the status indicator, such as "device is restarting", "device is in error state", "device is plugged in", "device battery is depleted", "detection device and smart communication device are attempting to connect", "detection device and smart communication device are attempting to connect", "device is preheating", "preheating completed", "test process is in progress" and / or "test process completed. Result analysis is in progress".

[0058] Test results, user prompts, etc. are preferably indicated via an application on the smart communication device and the display of the smart communication device.

[0059] According to a fourth aspect of the present invention, a method of operating a test system is provided.

[0060] The method of operating a test system preferably includes at least some of the following steps:

[0061] - providing a detection device according to one of the aspects described herein,

[0062] - providing a test container or test container assembly as described below,

[0063] - providing a smart communication device,

[0064] - activating the detection device,

[0065] - coupling the detection device to the smart communication device (if coupling is required; this step is therefore optional),

[0066] - Configure the detection device as follows:

[0067] - Read from the test container or test container component a code representing the ID of the determination or a script that identifies the test process and / or

[0068] defines the test process (this is preferred because the detection device can thus be a general-purpose device that can be adapted to individual test processes and determinations; in an alternative embodiment, the detection device is preconfigured for a specific test process suitable for a specific determination. Subsequently, there is no need to read the code from the test container or test container component. In an alternative embodiment of a general (i.e., programmable) detection device, the test process can be configured manually.)

[0069] - Upload a script that includes script commands defining the test process to be executed by the detection device, where the script and the test process thus defined correspond to the determination in the test container or test container component (again, this step

[0070] is optional; see above); and

[0071] - Place the test container of the test container or test container component in one or more detection chambers of the detection device (the order of the steps of configuring the detection device and placing the test container in the detection chamber is optional; however, it is preferred to first configure the detection device and then only place the test container in the detection chamber because it allows the test process to be automatically started by inserting the test container into the receptacle (detection chamber) of the detection device).

[0072] - Once the test container of the test container or test container component is placed in the detection chamber, automatically start the test process defined by the uploaded script (automatic start is preferred; in an alternative embodiment, starting the test process is manually triggered, for example, via a graphical user interface on a smart communication device),

[0073] - Store the parameter values obtained during the test process in the memory of the detection device,

[0074] - Read, via a smart communication device, the parameter values obtained during the test process from the memory of the detection device.

[0075] - Upload the parameter values obtained during the test process to the server.

[0076] The steps of reading and outputting the parameter values and uploading the parameter values to the server are optional because at the end of the corresponding test process, the test result is usually immediately indicated to the user, for example, by the state of the light indicating the detection device or a message on the smart communication device.

[0077] In the case where the test container or test container component is provided with an ID code that can be read out by the detection device itself, the configuration of the detection device and the start of the test process can be fully automated. For example, the test container or test container component can be provided with an RF-ID chip, which can be read out by the NFC interface of the detection device when the test container or test container component is placed in the receptacle of the detection device. Then, once the test container or test container component is placed in the detection chamber, the configuration of the detection device and the start of the test process can be automatically carried out.

[0078] Preferably, the parameter values obtained during the test process are analyzed by the server. In a particularly preferred embodiment, the server is configured to perform a cluster analysis on the parameter values obtained during the test processes of different detection devices.

[0079] The method preferably further includes the step of encrypting the obtained parameter values stored in the memory.

[0080] The method preferably further includes the step of deleting the script commands in the memory once the test process is completed.

[0081] The test container component according to the fourth aspect may include a single cracking chamber, a plurality of test containers, and a metering device. The metering device is selectively fluidly connected to the cracking chamber and includes a plurality of metering compartments for metering and simultaneously transferring the same portion of the cracked sample from the cracking chamber into the test containers.

[0082] The metering device may include:

[0083] - a volumetric metering compartment, and / or

[0084] - a microfluidic sample dispensing device.

[0085] Thus, quantitative dispensing of the cracked sample liquid can be achieved.

[0086] - a volumetric metering assembly, and / or

[0087] - through microfluidic sample distribution and metering.

[0088] Each metering compartment of the metering assembly preferably has at least one inlet opening and at least one outlet opening. The inlet opening and the outlet opening can be selectively closed and opened. The metering assembly includes a manually operable control device for selectively opening and closing the inlet opening and the outlet opening.

[0089] The metering assembly is preferably integrated in a container assembly including the cracking chamber.

[0090] The metering assembly may include a rotatable disk. In one rotational position of the disk, the metering compartment is open towards the lysis chamber, and in a different rotational position, the metering compartment has an open outflow opening for releasing the contents of the metering compartment towards different test vessels.

[0091] In the test system, the number of metering compartments preferably corresponds to the number of detection chambers of the detection device.

[0092] If, for example, four detection chambers are provided and thus four test vessels, the disk-shaped member of the metering assembly may be configured to rotate by slightly less than a quarter of a full turn.

[0093] In one embodiment, the metering compartment is additionally fluidly connected to the lysis chamber, i.e., the metering compartment is open towards the lysis chamber. To achieve a uniform distribution of the sample in the lysis chamber and the metering compartment, the metering compartment preferably has a large opening towards the lysis chamber.

[0094] In an alternative embodiment, the metering compartment is initially fluidly separated from the lysis chamber. After lysis, the metering compartment is fluidly connected to the lysis chamber. For example, a rotating or otherwise movable part of the metering assembly will open the fluid connection between the lysis chamber and each metering compartment. For example, the lysis chamber may rotate about the metering compartment to a position where the outflow port of the lysis chamber is aligned with the inflow opening of the corresponding metering compartment.

[0095] According to different preferred embodiments, the rotation may be caused by manual actuation or by a motor in the test device. If four metering compartments are provided, the rotation is preferably a quarter turn.

[0096] A further rotation will close the fluid connection between the lysis chamber and the metering compartment, and an even further rotation will open the outflow opening of the metering compartment towards the test chamber.

[0097] The metering compartment may be a cavity and / or opening in a disk-shaped member of the metering assembly, which is placed between the bottom of the lysis chamber and the bottom of the container comprising the lysis chamber and the metering assembly.

[0098] In an alternative embodiment, the test vessel assembly may be similar to a syringe, where four metering compartments are arranged at the bottom of the syringe. Initially, these metering compartments are open towards the lysis chamber. Moving the piston towards the metering compartments will first close the metering compartments. Further moving the piston will then press the contents of the metering compartments into the test chamber.

[0099] Preferably, the metering device includes a flow control device configured for manual operation.

[0100] It has been found that reliable test results for different target analytes, especially different target nucleic acids, are facilitated if the sample is lysed once and the lysed sample is simultaneously dispensed into a plurality of test vessels containing test chemicals for different target analytes.

[0101] The system can be a point of care (POC) system, where the fluorescence detection device is arranged at the point of care, for example in a doctor's office. Alternatively, the system can be a personal system, where the fluorescence detection device is independent and portable, especially pocketable.

[0102] The test system and its components (i.e., the detection device, the intelligent communication device with the application installed thereon, and the test vessel assembly) allow for easy and fail-safe testing of analytes in a home and / or point of care environment and do not require specially educated personnel.

[0103] The invention will now be further illustrated by way of example and with reference to the accompanying drawings. In the drawings,

[0104] Figure 1 : the components of the test system are shown by way of example;

[0105] Figure 2 : is a schematic illustration of how a test vessel of the detection device and the test vessel assembly can be placed in a detection chamber formed by a receptacle of the detection device;

[0106] Figure 3 : is a schematic illustration of the test vessel assembly, which includes four test vessels (i.e., vials) containing a mixture of enzymes for detecting target nucleic acids;

[0107] Figure 4 : is a schematic illustration of the detection device;

[0108] Figure 5 : is similar to Figure 4 : is a schematic illustration of an alternative detection device of the test device, but each detection chamber has a single light source;

[0109] Figure 6 : is Figure 5 : is a schematic illustration of an alternative detection device of

[0110] Figure 7 : is Figure 5 : is a schematic illustration of an alternative detection device of

[0111] Figure 8 : schematically shows that the detection device can include a dedicated control module;

[0112] Figure 9 : Schematically shows the basic electronic components of a detection device that can be implemented as a control module;

[0113] Figure 10 a to Figure 10 c: Schematically shows alternatives for arranging a light source to illuminate a detection chamber and avoid a direct light path between the light source and the light sensor;

[0114] Figure 11 : Is a schematic flowchart showing a method of operating a test system.

[0115] Figure 12 : Shows a test container assembly that includes a lysis chamber and a metering assembly having four metering compartments for transferring a determined amount of fluid from the lysis chamber of each of four test containers;

[0116] Figures 13 to 16 Shows the operation of the metering assembly; and

[0117] Figure 17 Further shows the test container assembly.

[0118] A test system 10 for detecting a target analyte in a sample includes a detection device 12, one or more test containers 14, a smart communication device 16, and preferably a central server 18, which can communicate with a plurality of smart communication devices 16 and a plurality of detection devices 14; see Figure 1 .

[0119] The detection device 12 includes a plurality of receptacles 20 configured to receive the test containers 14; see Figure 2 . Each receptacle 20 defines a detection chamber of the detection device 12.

[0120] Preferably, the test container 14 is part of a test container assembly 22 that includes a plurality of test containers 14; see Figure 2 and Figure 3 . The test containers 14 of the test container assembly 22 are arranged in a fixed geometric constellation that matches the geometric constellation of the receptacles 20 of the detection device 12; see Figure 2 . Thus, all the containers 14 of the test container assembly 22 can be placed in the receptacles 20 of the detection device 12 simultaneously, allowing for example the simultaneous testing of samples for different target analytes. In particular, the detection device simultaneously tests samples contained in different test containers.

[0121] Detection device

[0122] The detection device 12 is configured to excite and detect luminescence in a sample contained in a test container 14 placed in a receptacle 20 of the detection device 12.

[0123] In Figures 4 to 8 a schematic view of the detection device 12, the basic components of the detection device 12 are shown: for each receptacle 20, an optical sensor 24 is provided which preferably can detect light in different wavelength bands. Each optical sensor 24 can be configured to detect the light intensity in a wide wavelength range or to distinguish different wavelength bands, thereby collecting spectral information.

[0124] Preferably, for each receptacle 20, a separate light source 26 is provided for irradiating the contents of the test container placed in the corresponding receptacle; see Figure 5 . Alternatively, a common light source 26' can be provided for all receptacles 20; see Figure 4 .

[0125] The optical light source 26 and the optical sensor 24 of each receptacle 20 are arranged to prevent the light emitted by the corresponding light source 26 from directly irradiating the corresponding optical sensor 24. In other words: there is no direct optical path between the light source 26 and the optical sensor 24. Thus, each optical sensor 24 only senses the light scattered by the sample in the test container or the luminescence generated in the sample in the test container.

[0126] Figure 10 a, Figure 10 b and Figure 10 c show in more detail below different preferred arrangements of the light source 26 and the optical sensor 24.

[0127] Each light source 26 is configured to emit light in different wavelength bands. In this wavelength band there is a wavelength band which, if the light in this wavelength band irradiates the sample, can excite luminescence in the sample contained in the test container 14.

[0128] Light in additional wavelength bands allows the detection of, for example, the turbidity of the sample in the test container 14.

[0129] Furthermore, a heating device 28 is provided for heating the wall of the receptacle 20. Preferably, the receptacle 20 is arranged in a common heated metal block 30. The metal block 30 provides a uniform heat distribution and thus a uniform temperature in all receptacles 20. The heating device 28 is preferably a temperature-controlled electric heating device. Preferably, a feedback temperature control connected to at least one temperature sensor 32 is provided. The temperature sensor 32 is preferably arranged at the center of the metal block 30 surrounding the receptacle 20.

[0130] Each receptacle 20 defines a detection chamber which can be heated and irradiated to excite and detect luminescence.

[0131] In an alternative embodiment (not shown), a separate heating device can be provided for each receptacle 20 (i.e., for each detection chamber), thereby reducing the thermal mass to be heated by a single heating device.

[0132] Preferably, a common thermal mass provided by the metal block 30 is provided. The thermal mass (and thus the heat capacity) of the metal block 30 is preferably at least ten times the heat capacity of the sample fluid in all the test containers 14 placed together in the receptacle 20.

[0133] The electrical components described so far (i.e., the light sensor 24, the light source 26, the heating device 28, and the temperature sensor 32) are connected to the controller 40 and the power supply 42. The power supply 42 is preferably a rechargeable battery. The controller 40 and the power supply 42 are preferably connected to the control module 44 or a part of the control module 44, and the control module 44 is electrically connected to the electrical components 24, 26, 28, and 32 through a flexible electrical conductor (such as a flexible printed circuit board (flexible PCB) or flexible wire), thereby restricting the heat transfer between the control module 44 and the electrical components (i.e., the light sensor 24, the light source 26, the heating device 28, and the temperature sensor 32) arranged around the receptacle 20.

[0134] The controller 40 is also connected to the memory 46 and the wireless data transmission interface 48. The controller 40 is also connected to the light source drive circuit 50, the light sensor sub - controller 52, and the heating device feedback controller circuit 54. The controller 40 is connected to the memory 46, the wireless data communication interface 48, the light source drive circuit 50, the light sensor sub - controller 52, and the heating device feedback controller circuit 54 via a data bus 56 (preferably an I2C bus). The controller 40, the memory 46, the wireless data communication interface 48, the light source drive circuit 50, the light sensor sub - controller 52, and the heating device feedback controller circuit 54 are preferably arranged on a common printed circuit board (PCB, main board) as part of a controller module. The light source 26, the light sensor 24, the heating device 28, and the temperature sensor 32 are preferably connected to the main board through a flexible circuit board or flexible wires.

[0135] The heating device 32 preferably includes a conductor arranged in a meandering shape or a spiral shape and serving as a heating wire.

[0136] The light source drive circuit 50 is configured to be able to control the light source 26 through the controller 40. Controlling the light source includes controlling the intensity of the emitted light in one or more wavelength bands. In the simplest case, the control of the light source is a simple on - off switching of the light source 26.

[0137] The light sensor sub - controller 52 is configured to enable the corresponding light sensor 24 to be read out by the controller 40, thereby obtaining a light sensor output signal that can be processed by the controller 40.

[0138] The heating device feedback controller circuit 54 is configured to control the heating device 28 and is controlled by the controller 40. The heating device feedback controller circuit 54 is electrically connected to at least one heating device 28 for heating the receptacle 20. The heating device feedback controller circuit 54 may also be connected to the temperature sensor 32. The heating device feedback controller circuit 54 can implement feedback temperature control to heat the receptacle to the temperature set by the controller 40 and control the temperature. Alternatively, the heating device feedback controller circuit 54 can provide an interface for connecting the temperature sensor to the controller 40 and driver electronics for controlling the heating device 28 through the controller 40. Then, the temperature feedback control can be defined by a temperature control software program stored in the memory 46.

[0139] The memory 46 includes at least a detection device operating system, that is, control software executed by the controller 40 during the operation of the detection device 12.

[0140] In addition, a status light controlled by the controller 40 via the LED driver circuit 38.1 is provided, preferably an RGB light-emitting diode (LED) 38. The controller 40 is configured via the device operating system to make the LED indicate different light codes, such as making the LED light up in different colors or in different blinking codes. Each light code represents a predefined state or different events of the detection device.

[0141] Except for the USB terminals and the opening of the receptacle 20, the housing 34 of the detection device is preferably completely closed; see Figure 6 . In addition, the status indicator light 38.1 is visible from the outside. A closure (not shown) for the opening of the receptacle 20 can be provided to prevent light from entering the receptacle when no test container is placed in the receptacle or some receptacles 20.

[0142] The operating system stored in the memory 46 includes at least the following software components:

[0143] - Basic autonomous device operating system

[0144] - Temperature feedback control program

[0145] - Self-test program

[0146] - A script interpreter for script commands received from a smart communication device, and

[0147] - Data communication program

[0148] Via the wireless data transfer interface 48, the detection device 12 can communicate with an intelligent communication device 14 such as a smart phone. An application (i.e., software program) is installed on the intelligent communication device 16, which enables communication with the detection device 12 and provides a user interface for controlling the detection device 12 and for displaying information about the detection device 12 and information received from the detection device 12 to the user. The user interface of the intelligent communication device 16 is preferably a graphical user interface.

[0149] According to a preferred embodiment, functions provided independently by the detection device operating system and the controller 40 of the detection device 12 from the functions provided by the intelligent communication device 16 are the following functions:

[0150] - Power on and power off

[0151] - Self-test

[0152] - Script interpreter

[0153] In addition to this, the application installed on the intelligent communication device 16 provides the following additional functions:

[0154] - Detection device configuration via scripts and script commands

[0155] - Detection device software and firmware updates

[0156] - User interface with prompts for the user

[0157] - Control of the detection device via script commands during the test process

[0158] - Read data from the detection device memory 46

[0159] - Communicate with the server 18

[0160] Due to the functional separation between the detection device 12 and the intelligent communication device 16, the detection device can be implemented as a general-purpose low-cost device, which provides the basic functions for luminescence excitation and detection in a sample.

[0161] With the aid of the intelligent communication device 16 and / or the server 18, the detection device 12 can be remotely configured for different test processes. In particular, parameters for different test processes can be stored in the server 18 and / or in the intelligent communication device 16, such that they can be sent to the detection device 12. The latter aspect is supported by the script interpreter installed on the detection device 12. Thus, the detection device 12 can interpret scripts and operate according to script commands. The script interpreter is configured to interpret a limited number of script commands. This is for security reasons, because the detection device 12 cannot be freely programmed. Instead, the script commands in combination with the script interpreter are configured to ensure the operation of the detection device 12 within the design limits.

[0162] With the aid of script commands, entire test procedures can be defined such that they can be executed by the detection device 12. In particular, different test procedures for different target analytes can be provided and defined via the script.

[0163] Method and test procedure for operating a test system:

[0164] Generally, the method for operating a test system and test procedure includes at least some of the following steps:

[0165] First, provide the detection device 12, the intelligent communication device 16, and the test component 22.

[0166] The detection device may need to be charged. For charging, the detection device 12 is connected to a typical charging device.

[0167] Once the detection device 12 is charged, it can be switched on. Then, the detection device 12 performs a self-test. According to the preferred embodiment, all receptacles (detection chambers) 20 of the detection device are covered during the self-test.

[0168] At the end of the self-test, the detection device is ready to be connected to the intelligent communication device 16. The readiness of the detection device 12 is indicated by a color status light (e.g., RGB LED 38). The wireless data connection between the intelligent communication device 16 and the detection device 12 can be initiated by arranging the intelligent communication device 16 at a short distance from the near-field communication chip 48.1 of the detection device 12. In the memory of the NFC chip 48.1, a Bluetooth connection ID is stored, which is then sent to the intelligent communication device and enables the intelligent communication device to establish a Bluetooth data communication between the intelligent communication device 16 and the detection device 12. Accordingly, the Bluetooth connection between the intelligent communication device 16 and the detection device 12 is established via a tab-to-connect.

[0169] In addition, on the memory of the NFC chip 48.1, a link for an application to be installed on the intelligent communication device 16 can be stored. The link can also be transmitted to the intelligent communication device 16 via near-field communication. Thus, the application for operating the detection device 12 can be easily installed on the intelligent communication device 16.

[0170] The application also enables the intelligent communication device 16 to connect to the server 18.

[0171] To configure the detection device 12 to perform a specific test procedure, the intelligent communication device 16 is used to read a code from the test container component 22. The code to be read can be a graphical code, such as a QR code or a bar code or any other kind of matrix code. The code can also be a code stored on an NFC chip attached to the test container component 22. The code at least contains the concept of one or more measurements provided by the test container 14 of the test container component 22. Based on the code read from the test container component 22, an application running on the intelligent communication device 16 initiates the transfer of a script defining a suitable test procedure from the server 18 to the detection device 12. In other words, for each measurement or each combination of measurements, a specific test procedure is defined. To configure the detection device 12 to perform a specific test procedure, the script is stored on the server 18 or on the intelligent communication device 16, which can be interpreted by a script interpreter of the detection device and configure the detection device 12 to perform the steps of a single test procedure.

[0172] First, no test procedure is defined on the detection device. For safety reasons, after the test procedure is completed, the script defining the test procedure is not stored on the detection device 12.

[0173] The script defines, for example, the timing of the steps of the test procedure, the values to be measured, the control of the light source 26, the storage of the measurement parameter values, and the triggering of a message shown to the user on the display of the intelligent communication device 16. The messages to be shown to the user are defined by an application installed on the intelligent communication device 16. The display of these messages can be initiated by a command received from the detection device 12 during the test procedure.

[0174] The script commands can also define the color and / or the blinking code of the data slide 38 during different phases of the test procedure. For example, a dedicated color and / or blinking code of the data slide 38 can indicate the end of the test procedure to the user independently of the message shown to the user on the display of the intelligent communication device 16.

[0175] Parameter values measured by the light sensor 24 of the detection device 12 during the test procedure, for example, and other parameters, such as the temperature value during the test procedure or the light intensity during the test procedure, are stored in the memory 46 of the detection device 12. Preferably, all the stored parameter values are encrypted.

[0176] Once the test procedure is completed, the parameter values stored during the test procedure can be read out by means of the intelligent communication device 16 and can be sent to the server 18. However, the parameter values are stored in the memory 46 of the detection device 12 together with the test procedure ID or the measurement ID as long as these parameter values are not read out by the intelligent communication device 16. In other words, it is not necessary to read out the stored parameter values during or immediately after the test procedure. Instead, the parameter values can be read out later.

[0177] The script interpreter and the detection device operating system are part of the firmware of the detection device 12. The firmware of the detection device 12 can also be updated by means of the intelligent communication device 16 and the application installed thereon.

[0178] All test procedures performed by the detection device 12 are preferably configured to take no more than 15 minutes, preferably no more than 10 minutes or even less than 5 minutes. The duration of the test procedure can be optimized because the test procedure is always individually adapted to a specific determination. Thus, the combination of a specific determination contained in the test container 14 and the optimized test procedure for each determination provides a short test procedure.

[0179] The detection device 12 in combination with the intelligent communication device 16 or the server 18 is configured for central management of a plurality of detection devices by means of a central server. The central server 18 allows immediate quality assessment by comparing the data received from different detection devices 12. In addition, the data measured by a single detection device 12 can be compared with the data measured by other detection devices 12 applying the same test procedure. This allows a cluster evaluation of the data collected by different devices.

[0180] In addition, this enables implicit control of the test because the test results (data obtained by the detection device 12) can be compared with reference data collected by other detection devices. In the case where the measured data deviates significantly from the reference data stored on the server 18, a warning can be generated indicating to the user that the test may have failed.

[0181] Since the data generated by the detection device 12 can be sent to the intelligent communication device 16 and / or the central server 18, a test history log can be generated and stored.

[0182] In addition, the application (app) on the intelligent communication device is preferably configured to provide feedback to the user during use. In particular, the application can be configured to indicate to the user step by step all the manual steps that the user has to perform during the test procedure.

[0183] The intelligent communication device 16 can also be configured to generate a note at the end of the test procedure to remind the user to handle the sample after the test in the correct manner.

[0184] On the other hand, it also relates to the test container 14 and the chemicals contained therein. Indications of the chemicals contained in the test container 14 and, for example, their production dates can be attached to each test container component 22 by means of a graphical code (such as a QR code or a bar code, etc.). Such a code can be read out by the intelligent communication device 16. The intelligent communication device 16 can then transmit a script to the detection device 12 for configuring the detection device 12 in a manner suitable for the chemicals in the test container 14 of the test container component 22.

[0185] Other sensors of the detection device 12 not mentioned are a position sensor or an attitude sensor, a humidity sensor, and a temperature sensor for the ambient temperature. The ambient temperature can also be indirectly determined during the heating of the detection device 12 by measuring the energy required to heat the detection device 12 to a specific temperature.

[0186] The system 10 is preferably a decentralized system having a plurality of independent detection devices 12. Each detection device 12 is a reusable low-cost device that facilitates multiple uses with low-cost disposable items. The low-cost disposable items can be, for example, test container components such as the test container component 22.

[0187] Preferably, the detection device 12 does not have buttons and can be fully controlled via an external device such as the intelligent communication device 16. This makes the detection device 12 more robust and avoids contamination. Except for the USB terminal and the opening of the receptacle 20, the housing 34 of the detection device 12 is completely closed.

[0188] Typical methods of operating the test system include at least some of the following steps:

[0189] - Power on the detection device 12, preferably initiated by the user with the help of the intelligent communication device 16,

[0190] - Start the detection device 12, turn on the sensors and actuators (heating device and light source), start the device OS, and start the script interpreter

[0191] - Automatic self-test of the detection device 12,

[0192] - Connect the detection device to the intelligent communication device 16,

[0193] - Read the code representing the measurement ID from the test container component by means of the intelligent communication device 16,

[0194] - Load a script defining the test process from the server via the external intelligent communication device 16, and the test process is configured to be suitable for the measurement contained in the test container,

[0195] - Upload the script including the script commands to be interpreted by the script interpreter to the detection device memory 46,

[0196] -Execute a test process according to the script commands received from the server 18 and stored in the memory 46.

[0197] -Heat (automatically, without user interaction) the detection chamber 20 of the detection device 12.

[0198] -Prompt the user to insert the test container 14 into the receptacle 20 (prompt via the external intelligent communication device 16).

[0199] -Automatically detect the inserted test container 14, for example, by detecting an event in the output signal of the optical sensor.

[0200] -Start lighting and measurement according to the process defined by the script stored in the memory 46 of the detection device 12.

[0201] -Store the sensed optical parameter values obtained from the output signal of the optical sensor.

[0202] -Check data integrity.

[0203] -Observe user interaction.

[0204] -Generate a concurrent signal to notify a warning message in case of a faulty or incorrect user interaction (e.g., removing the test container or incorrect posture of the detection device).

[0205] -Indicate sensed / non-sensed luminescence to the user via the status indicator 38.1 of the detection device (less preferred) or via a message displayed on the display of the intelligent communication device 16 (preferred) or both.

[0206] -Notify the user of the successfully completed test process or the failed test respectively.

[0207] -Read out the sensed optical parameter values stored in the memory 46 of the detection device 12.

[0208] -Send data representing the optical parameter values to the server 18 via the intelligent communication device.

[0209] -Evaluate the data representing the optical parameter values.

[0210] As Figure 11 The method of operating a test system as shown includes the following steps:

[0211] -Provide a detection device according to one of the detection devices described herein.

[0212] -Provide a test container or test container assembly as described below.

[0213] -Provide an intelligent communication device.

[0214] - Activate the detection device (S1),

[0215] - If necessary, connect the detection device to the intelligent communication device (S2),

[0216] - Configure the detection device or the test process through the following process (S3):

[0217] - Read the code from the test container or the test container component (S3.1),

[0218] - Upload a script that defines the script commands of the test process to be executed by the detection device. The script and the test process defined thereby correspond to the determination in the test container or the test container component (S3.2), and

[0219] - Place the test container of the test container or the test container component in one or more detection chambers of the detection device (S4),

[0220] - Once the test container of the test container or the test container component is placed in the detection chamber, automatically start the test process defined by the uploaded script (S5),

[0221] - Store the parameter values obtained during the test process in the memory of the detection device (S6),

[0222] - Read the parameter values obtained during the test process from the memory of the detection device by means of the intelligent communication device (S7), and

[0223] - Upload the parameter values obtained during the test process to the server (S8).

[0224] Figure 11 The steps in the boxes defined by the dashed lines in [] are optional or preferred respectively.

[0225] According to a preferred embodiment, the detection device may have other sensors in addition to the optical sensor 24. As another optional sensor, a temperature sensor 32 is preferably provided because the temperature sensor 32 realizes feedback control of the temperature. Other sensors that are not shown in the figure and may be implemented together with the detection device 12 are an inertial measurement unit for determining the orientation or attitude of the detection device 12 and a humidity sensor that can sense the humidity of the air in the environment of the detection device 12. The output signals generated by these sensors are preferably also stored in the memory 46 of the detection device 12.

[0226] Preferably, all data representing the measured parameter values and / or events are sent to server 18. In a preferred system, multiple detection devices 12 can be managed by one server (or a group of servers) 18. Thus, server 18 is capable of analyzing data representing the measured parameter values from different detection devices 12. Since the data received by server 18 during or after the test process contains not only data representing the measured parameter values, but also a certain determination ID, server 18 can be adapted to analyze the measured corresponding parameter values from different detection devices related to the same kind of determination. Therefore, the test process for each determination can be further optimized and a script with script commands can be generated accordingly. The optimized test process for each determination represented by a specific script minimizes the risk of failure testing or incorrect user manipulation. In addition, the adaptation of the test process to each specific determination enables a short test process. For example, the test process is shorter than 15 minutes, preferably shorter than 10 minutes or even shorter than 5 minutes.

[0227] The script interpreter of the detection device defines a detection device-specific script language. The script language is designed to limit the effects of script commands, for example, regarding controlling sensors of the detection device or heating devices and / or light sources, such that all components will always operate within their specific design limits. Thus, improved device safety is achieved through limited script commands.

[0228] Server 18 or a group of servers 18 includes a database, in which, for each determination, a specific test process and a script corresponding to the specific test process are stored. Thus, once the intelligent communication device requests the script for a specific determination ID, the specific script can be easily downloaded from the server.

[0229] Providing a central server 18 also has the advantage of allowing central management of all detection devices and performing continuous quality assessment. Data representing the measured parameter values received from different detection devices can be analyzed through cluster evaluation, which further improves the sensitivity and specificity of the test defined by each specific test process.

[0230] The data generated by server 18 can be available to applications installed on a specific intelligent communication device. Thus, the intelligent communication device can perform implicit control of each test when connected to the detection device during the test.

[0231] However, since the detection device 12 has sufficient memory 46, in the case where the data communication between the detection device 12 and the intelligent communication device 16 is interrupted during the test process, the detection device 12 can also operate autonomously. The measured parameter values stored in the memory 46 of the detection device 12 can be read out at any time later, for example, several hours or days after the test process is completed.

[0232] The application installed on the intelligent communication device 16 is preferably configured to provide user feedback during each test process. In particular, the application is configured to provide step-by-step instructions to the user and to notify the user of a specific state of the detection device or the result of a test performed by means of the detection device. The application installed on the intelligent communication device 16 may also prompt the user to set the test container component 22 in a safe manner at the end of the test.

[0233] As already pointed out, the parameter values measured during the test process include temperature. The temperature value is generally related to the temperature in the detection chamber 20. However, considering the power consumed by the heating device 28 and the temperature values measured during heating, the ambient temperature can also be determined by analyzing the time course of the temperature values relative to the power consumed by the heating device. Therefore, instead of providing a dedicated temperature sensor for the ambient temperature, the ambient temperature can be determined indirectly by means of the temperature sensor for the detection chamber 20.

[0234] Preferably, the power requirement of the detection device 12 is less than 10 W (watts). The weight of the detection device is preferably less than 250 grams, even more preferably less than 100 g or less than 50 g. Preferably, the overall volume of the detection device is less than 250 cm 3 .

[0235] Figure 10 a, Figure 10 b and Figure 10 c show ways to prevent the light emitted by the light source 26 from directly irradiating the light sensor 24 of the corresponding receptacle 20.

[0236] As Figure 10 shown in a, the light source 26 and the light sensor 24 can be laterally arranged with respect to the receptacle 20 at an angle that prevents the light emitted from the light source 26 from directly irradiating the corresponding light sensor 24. This angle can be, for example, 90°. The wall (e.g., the wall of the metal block 30) has a lateral hole 36.1 that guides the light from the light source 26 into the receptacle 20 and prevents direct irradiation of the light sensor 26.

[0237] Figure 10 Another preferred arrangement of the light source 26 and the corresponding light sensor 24 is shown in b. Thus, the light source 26 and the light sensor 24 are arranged one above the other on the same side of the receptacle 20. Thus, direct irradiation of the light sensor 24 by the light source 26 is avoided.

[0238] Alternatively, as Figure 10 shown in c, the light source 26 and the light sensor 24 can be placed on the same horizontal plane, for example, on a printed circuit board, such as a daughter board for the light sensor and the light source, which is flexibly connected to a main board that carries a controller 40, etc. Then, the light from the light source 26 can be guided by an optical waveguide 36.2 and laterally fed into the receptacle 20. However,Figure 10 An embodiment of a or Figure 10 b is preferred because the lateral arrangement of the light source 26 and the light sensor 24 is less affected by particles deposited on the bottom of the test container.

[0239] Test container assembly with a metering device

[0240] Figures 12 to 17 A test container assembly 22 with a metering device 60 according to a second aspect of the present invention is shown. The test container assembly 22 includes a single lysis chamber 62 and a metering device 60 for dispensing a determined amount of fluid from the lysis chamber 62 into respective test containers 14. The lysis chamber 62 contains a liquid lysis fluid that causes lysis of cells in the sample, thereby releasing nucleic acids (DNA or RNA). The lysis fluid may include an acid (such as HCl or a weak base) and a surfactant.

[0241] Each test container 14 contains a chemical mixture that can cause nucleic acid amplification in the sample. Preferably, the mixture includes a target-specific probe and enzymes, in particular recombinase, single-stranded DNA-binding protein (SSB), and a strand-displacement polymerase that causes recombinase polymerase amplification (RPA). The test container also preferably contains exonuclease III, allowing real-time fluorescence detection using an exonuclease probe. The mixture may be provided in the form of dry granules 64.

[0242] According to a preferred embodiment, one of the containers 14 contains a reference or control assay that will always cause luminescence if the test system is properly processed and the test procedure is performed without failure.

[0243] Each metering compartment 66 of the metering assembly 60 preferably has at least one inlet opening and at least one outlet opening. The inlet and outlet openings can be selectively closed and opened. The metering assembly includes manually operated control means for selectively opening and closing the inlet and outlet openings.

[0244] The metering device is preferably integrated in the container assembly 22 including the lysis chamber 62.

[0245] The metering device may include a rotatable metering disk 68. In one rotational position of the metering disk 68, the metering compartment 66 is open towards the lysis chamber 62, and in a different rotational position of the metering disk 68, the metering compartment 66 has an open outlet opening for releasing the contents of the metering compartment 66 towards different test containers 14.

[0246] The number of metering compartments 66 corresponds to the number of test containers 14 of the test container assembly 22.

[0247] If, for example, four detection chambers 20 are provided and thus four test containers 14 are provided, the metering disk 68 of the metering assembly 60 can be configured to rotate slightly less than a quarter of a full turn.

[0248] In one embodiment, the metering compartments 66 are additionally fluidly connected to the lysis chamber 62, i.e., the metering compartments open towards the lysis chamber 62. To achieve a uniform distribution of the sample in the lysis chamber 62 and the metering compartments 66, it is preferred that the metering compartments 66 each have a large opening towards the lysis chamber 62.

[0249] In an alternative embodiment, the metering compartments are initially fluidly separated from the lysis chamber. After lysis, the metering compartments are fluidly connected to the lysis chamber. For example, a rotating or otherwise movable part of the metering assembly will open the fluid connection between the lysis chamber and each metering compartment. For example, the lysis chamber can be rotated relative to the metering compartments to a position where the outflow port of the lysis chamber is aligned with the inflow opening of the corresponding metering compartment.

[0250] According to different preferred embodiments, the rotation can be caused by manual actuation or by a motor in the test device. If four metering compartments are provided, the rotation is preferably a quarter turn.

[0251] A further rotation will close the fluid connection between the lysis chamber and the metering compartments, and an even further rotation will open the outflow opening of the metering compartments with respect to the test chamber.

[0252] The metering compartments 66 can be cavities and / or openings in a disk-shaped member of the metering assembly, and the disk-shaped member is placed between the bottom of the lysis chamber and the bottom of the container including the lysis chamber and the metering assembly.

[0253] Figures 12 to 17 A test container assembly 22 with a metering device is shown. Figures 12 to 17 The test container assembly 22 includes a lysis chamber 62, a metering device 60, and four test containers 14.

[0254] The metering device 60 includes a metering disk 68 and four sliding metering pistons 70.

[0255] By rotating the lysis chamber 62 relative to a bottom plate 72 having an outlet port 74, the metering disk 68 and the sliding metering pistons 70 can be rotated or moved respectively, and the outlet port 74 is fluidly connected to the internal space of the test container 14; Figure 12 a is an exploded perspective view of the test container assembly 22. Figure 12 b is a top view of the test container assembly 22, Figure 12 c is a bottom view of the test container assembly 22.

[0256] Figures 13 to 16 Shows the steps for transferring a determined amount of fluid from the cracking chamber 62 to each test chamber 14.

[0257] Initially, the metering compartment 66 and the metering tray 68 are fluidly connected to the interior of the cracking chamber 62, as Figure 13 shown in FIGS. Figure 13 a-e. The bottom of the cracking chamber 62 is positioned such that an opening 74 in the bottom of the cracking chamber 62 is aligned with a cavity in the metering tray 68, thereby forming the metering chamber 66. Thus, fluid from the interior of the cracking chamber 62 can flow into the metering compartment 66 and fill the metering compartment 66. Each metering compartment has a volume of, for example, 50 μl. In alternative embodiments, the metering compartments can each have a capacity between 20 μl and 100 μl.

[0258] In the next step, as shown in FIGS. Figure 14 a-d, the metering compartment 66 is closed to prevent further fluid from flowing from the interior of the cracking chamber 62 into the metering compartment 66. Closing the metering compartment towards the cracking chamber 62 is achieved by rotating the cracking chamber 62 counterclockwise (if viewed from above) to the position shown in FIG. Figure 14 a. The metering tray 68 and the piston 70 each remain in their initial positions, as shown in FIGS. Figure 14 b and Figure 13 b, respectively. Similarly, the bottom plate 72 does not move at all; see FIGS. Figure 14 c and Figure 13 c. Figure 14 c.

[0259] If the cracking chamber 62 is further rotated counterclockwise, the metering tray 68 and the piston 70 also rotate counterclockwise until the outlet port 76 of the metering compartment 66 opens to the inlet channel 78 of the test container 14 in the bottom plate 72; see FIGS. Figure 15 a-d. Again, the bottom plate 72 does not move as seen in FIGS. Figure 15 c, Figure 13 c, Figure 14 c, and Figure 15 c, but rather both the cracking chamber 62 and the metering tray 68 with the piston 70 rotate about their previous positions (shown in FIGS. Figure 13 a and Figure 13 b); see FIGS. Figure 14 a and Figure 14 b.

[0260] Finally, as shown in FIGS. Figure 16 a- Figure 16As shown in Figure 16 a, and Figure 16 b, the contents of the metering compartment 66 are extruded from the metering compartment 66 by means of the sliding piston 70 and into the test vessel 14. This is achieved by further rotating the lysis chamber 62 counterclockwise. This rotation of the lysis chamber has the effect of pushing the sliding piston in the counterclockwise direction while the metering disk 68 does not rotate further; see

[0261] Figure 17 Similar to Figure 12 a, and shows the bottom plate 72 of the test vessel assembly 22 interconnected to four test vessels 14, each test vessel including dry granules having chemicals and biochemicals (especially enzymes).

[0262] The test vessel assembly 22 described herein is particularly suitable for use with the detection device 12 as described herein, because the test vessel assembly 22 facilitates the simultaneous initiation of different chemical and / or biochemical reactions in the test vessels 14, thereby allowing an easy comparison of the time course of the signals generated by the sensors in the different detection chambers of the detection device, where the test vessels are placed in the detection device.

[0263] However, it should be noted that the detection device 12 can be used independently of the test vessel assembly 22 described herein, and vice versa. In particular, the detection device 12 can be used with other test vessel assemblies or even with individual test vessels. The number of test vessels placed in the receptacle 20 of the detection device can even be less than the number of receptacles. Thus, samples in a single vial can even be analyzed with the detection device.

[0264] The fluorescence detection device 12 includes a plurality of detection chambers 20, for example four detection chambers, which fit closely with the test vessels 14 of the test vessel assembly 22. Each detection chamber 20 is configured to receive a test vessel 14. Inside or adjacent to the respective detection chamber 20, a light source 24 and a light sensor 26 are arranged. The light source 24 is configured to irradiate the contents of the respective detection chamber 20 with light, which can cause luminescence in the sample to be tested during and after the sample undergoes recombinase polymerase amplification. The light sensor 24 is arranged and configured to detect the luminescence in the detection chamber 20 in the case where luminescence occurs. For each detection chamber 20, a separate light sensor 26 is provided. To irradiate the contents of the test vessels 14 placed in different detection chambers 20, a common light source 26' can be provided: alternatively, a separate light source 26 can be provided for each detection chamber.

[0265] The detection chambers 20 are preferably arranged equidistantly in a rotationally symmetric manner.

[0266] Figure 9 The basic electronic components of the detection device 12 are shown. To power the light source 26 and the light sensor 24, an energy source 42 is provided, see Figure 9 . The energy source 42 may include a battery, preferably a rechargeable battery. Alternatively or in addition, the energy source 42 may include a power interface for connecting the fluorescence detection device 12 to an external power supply. The power interface may be a wired connection or wireless. The energy source 42 may also include a solar cell for providing a photovoltaic power supply.

[0267] The light source 24 and the light sensor 26 are also connected to a controller 40, which is configured to control the operation of the light source 24 and the light sensor 46 and further to read out the sensor output signal provided by the light sensor 26. The controller 40 may be a microcontroller or a state machine.

[0268] The controller 40 is operably connected to a wireless data interface 48, which is configured to allow data communication between the microcontroller 40 and an external device, such as a mobile phone or another device for data communication and data processing.

[0269] Preferably, the wireless data interface 48 is operably connected to the controller 40, the energy source 42, and the data memory 46, and is configured to provide energy harvesting, data storage, and data communication. In particular, the wireless interface 48 implements means for near field communication (NFC) and includes a data bus for communicating with the controller 40, such as an I2C data bus 56. The wireless data communication interface 48 is preferably configured to allow two-way data communication in order to transmit data generated by the fluorescence detection device 12 to an external device and to receive control commands and / or software updates from an external device (e.g., a smart communication device), such that the fluorescence detection device 12 can be controlled and updated by means of the external device.

[0270] The wireless data interface 48 may also implement WIFI communication as an alternative to near field communication. Another alternative is Bluetooth communication.

[0271] The wireless data communication interface 48 configured for data transmission via NFC or RFID preferably includes one or more antennas 58 serving as radio-frequency (RF) interfaces for transmitting electromagnetic signals representing digital data to one or more additional antennas of an external device by means of electromagnetic induction. The antenna 58 typically includes one or more coils, each coil having 4 or 5 windings.

[0272] The initiating party can be the intelligent communication device 16 that provides the carrier field, which is modulated by the data communication interface 48 for transmitting digital data. Preferably, in order to power the data communication interface 48, the data interface 48 draws energy from the intelligent communication device 16 via an NFC or RFID link. Thus, in particular, in the case where the transmitter unit is NFC or RFID enabled, the test device itself does not have to include an energy storage unit, such as a battery, for powering the data interface 48.

[0273] The detection device 12 is a single device that allows the test container 14 with the liquid sample as described above to be received. The evaluation of the presence of a specific analyte in the liquid sample is performed externally (e.g., directly on the intelligent communication device 16 that receives digital data from the detection device 12 via the wireless data communication interface 48). The intelligent communication device 16 can be a smart phone or a tablet computer, preferably having at least NFC or RFID capabilities and configured to be used as an initiating device. The intelligent communication device 16 can also be used to further transmit the digital data to, for example, a personal computer or a server 18 for evaluation purposes. Thus, the evaluation of the measurement data generated by the detection device 12 can be processed on one or more servers (i.e., the cloud). The evaluation of the measurement data preferably involves using a trained neural network on one or more servers.

[0274] The test container assembly 14 with the metering device 60 can be used to simultaneously dispense equal amounts of the lysed sample into the test container 14.

[0275] Reference numerals

[0276] 10 Test system

[0277] 12 Detection device

[0278] 14 Test container

[0279] 16 Intelligent communication device

[0280] 18 Server

[0281] 20 Receptacle

[0282] 22 Test container assembly

[0283] 24 Optical sensor

[0284] 26 Light source

[0285] 28 Heating device

[0286] 30 Metal block

[0287] 32 Temperature sensor

[0288] 34 Housing of the detection device

[0289] 36.1 Hole

[0290] 36.2 Light guide

[0291] 38 Status indicator (RGB LED)

[0292] 38.1 LED drive circuit

[0293] 40 Controller

[0294] 42 Power supply, rechargeable battery

[0295] 44 Control module

[0296] 46 Memory

[0297] 48 Wireless data transmission interface

[0298] 48.1 NFC transceiver

[0299] 48.2 Bluetooth transceiver

[0300] 50 Light source drive circuit

[0301] 52 Optical sensor sub - controller

[0302] 54 Heating device feedback control circuit

[0303] 56 Data bus; I2C bus

[0304] 58 Antenna

[0305] 60 Quantitative feeding device

[0306] 62 Pyrolysis chamber

[0307] 64 Dry granules with chemicals (enzymes)

[0308] 66 Quantitative feeding compartment, cavity of quantitative feeding tray

[0309] 68 Quantitative feeding tray

[0310] 70 Sliding piston arranged in the quantitative feeding compartment

[0311] 72 Bottom plate of the test container assembly

[0312] 74 Opening at the bottom of the pyrolysis chamber

[0313] 76 Outlet port of the quantitative feeding compartment

[0314] 78 Inlet channel for the test container

Claims

1. A test system (10), the test system comprising a detection device having a plurality of detection chambers (20) for receiving test containers (14) containing a sample to be tested and a chemical mixture including a target-specific probe and an enzyme, the target-specific probe and the enzyme being capable of causing nucleic acid amplification in the sample, the detection device further comprising a controller (40), a memory (46), and a data communication interface (48), the memory (46) and the data communication interface (48) being operatively connected to the controller (40), The test system further comprises a test container assembly (22) including a plurality of test containers (14) containing a sample to be tested and a chemical mixture including a target-specific probe and an enzyme, the target-specific probe and the enzyme being capable of causing nucleic acid amplification in the sample, The test system further comprises an intelligent communication device (16) configured for wireless data communication with the detection device (12).

2. The test system (10) according to claim 1, the test system comprising a detection device (12) according to any one of claims 3 to 10.

3. A detection device, the detection device comprising: A plurality of detection chambers (20) for receiving test containers (14); At least one light source (26) arranged and configured to irradiate at least one detection chamber (20) to cause luminescence in the sample to be tested, at least one light sensor (24) of each detection chamber (20) being configured to detect luminescence in the sample contained in the test container to be placed in the respective detection chamber (20); A controller (40); And a memory (46), Wherein the light sensor (24) and the at least one light source (26) are arranged to allow irradiation of the contents of the detection chamber by the at least one light source (26) and detection of the luminescence in the detection chamber (20) by the light sensor (24) assigned to the respective detection chamber (20), while preventing light emitted by the light source (26’) or the light source (26) from directly irradiating any light sensor (24), Wherein the controller (40) is at least indirectly connected to the light source (26) and the light sensor (24) for controlling the at least one light source (26) and for reading out the output value of the light sensor (24), Wherein the memory includes software defining the operation of the controller (40), And wherein the memory (46) is adapted to store at least parameter values corresponding to the output values of the light sensor (24).

4. The detection device according to claim 3, wherein, The memory includes software defining a script interpreter, and wherein the detection device is configured to receive script commands that, when interpreted by the script interpreter and executed by the controller, define a test procedure.

5. The detection device according to claim 3 or 4, wherein the detection device further comprises a heating device (28) for controlling the temperature in the detection chamber, the heating device being preferably configured to maintain a predetermined temperature in the detection chamber within a temperature range of + / - 0.5 K with respect to the predetermined temperature, and the heating device being controlled by a controller (40).

6. The detection device according to claim 5, wherein, The predetermined temperature is a temperature between 40 °C and 45 °C, preferably 42 °C.

7. The detection device according to any one of claims 3 to 6, wherein the detection device further comprises a metal block, the metal block being thermally coupled to the heating device and at least partially surrounding the detection chamber, the metal block being arranged and configured to provide a uniform heat distribution.

8. The detection device according to any one of claims 3 to 7, wherein the detection device further comprises a temperature sensor (32) for determining a temperature corresponding to the temperature in at least one of the detection chambers (20).

9. The detection device according to any one of claims 3 to 8, wherein the detection device further comprises a wireless data interface (48) for wirelessly transmitting data to and receiving data from a smart communication device (16).

10. The detection device according to any one of claims 3 to 9, wherein the detection device further comprises a status indicator (38) operatively connected to the controller (40).

11. A method of operating a test system according to claim 1 or 2, the method comprising the steps of: - providing a detection device (12) according to any one of claims 3 to 10, - providing a test container (14) or a test container assembly (22) according to any one of claims 14 to 15, - providing a smart communication device (16), - activating the detection device (12), - optionally coupling the detection device (12) to the smart communication device (16), - preferably configuring the detection device in the following manner - reading a code from the test container (14) or the test container assembly (22), - uploading a script, the script comprising script commands defining a test procedure to be executed by the detection device (12), the script and the test procedure defined thereby corresponding to the determination in the test container (14) or the test container assembly (22), and - placing the test container (14) of the test container (14) or the test container assembly (22) in one or more detection chambers (20) of the detection device (12), - automatically starting the test procedure defined by the uploaded script once the test container (14) of the test container (14) or the test container assembly (22) is placed in the detection chamber (20), - storing the parameter values obtained during the test procedure in the memory (46) of the detection device (12), - optionally reading out the parameter values obtained during the test procedure from the memory (46) of the detection device (12) by means of the smart communication device, and - Optionally upload the parameter values obtained during the test process to the server (18).

12. The method according to claim 11, wherein the method further comprises: The step of encrypting the obtained parameter values stored in the memory (46).

13. The method according to claim 11 or 12, wherein the method further comprises: The step of deleting the script commands in the memory (46) once the test process is completed.

14. A test container assembly (22), the test container assembly including a single lysis chamber, a plurality of test containers (14) selectively fluidly connected to the lysis chamber (62), and a metering device (60), wherein, The metering device (60) includes a plurality of metering compartments (66) for metering and simultaneously transferring the same portion of the lysed sample from the lysis chamber into the test container (14).

15. A test container assembly (22), the test container assembly including a single lysis chamber, a plurality of test containers (14) selectively fluidly connected to the lysis chamber (62), and a metering device (60), wherein, The metering device (60) includes a microfluidic sample dispensing device.

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