Method for testing a sample, in particular a biological sample, for specified sample parameters using a microfluidic system

Abstract

The invention relates to a method (500) for testing a sample for specified sample parameters using a microfluidic system (1000), in particular for detecting pathogens, wherein the microfluidic system (1000) is designed to test a biological sample for a set of various sample parameters, wherein one or more sample parameters are selected from the set, and the sample is tested for the selected sample parameters. The invention further relates to a microfluidic system (1000) for testing a sample for specified sample parameters, the microfluidic system being designed to carry out a method (500) according to the invention.

Description

[0001] Description

[0002] title

[0003] Method for testing a particular bi' Sample on with a microfluidic

[0004] State of the art

[0005] For medical diagnostic procedures that are to be carried out particularly at the point of care, for example assays for the detection of pathogens using nucleic acid amplification, microfluidic systems can be used, which are also referred to as lab-on-a-chip systems, since they automatically accommodate the functionalities of a macroscopic laboratory in integrated devices.

[0006] Such microfluidic systems can be designed to test a sample for a variety of parameters, depending on the application, for example, molecular diagnostic parameters relating to pathogens, resistance information, proteins or even clinical parameters such as gas values, amounts and concentrations of substances in the blood.

[0007] The microfluidic system can comprise a cartridge for receiving and processing the sample and an analysis device for receiving the cartridge and controlling the processing of the sample in the cartridge, as described, for example, in the documents DE 10 2016 222 075 A1 and DE 10 2016 222 072 A1. Disclosure of the invention

[0008] Advantages of the invention

[0009] Against this background, the invention relates to a method for testing a sample for predetermined sample parameters using a microfluidic system. The microfluidic system is designed to test a sample, in particular a biological sample, for a set of different sample parameters, wherein one or more sample parameters are selected from the set and the sample is tested for the selected sample parameters. The invention also relates to a microfluidic system designed in this way.

[0010] The microfluidic system can, as described above and without fundamentally limiting the invention, comprise a cartridge for receiving and processing the sample and an analysis device for receiving the cartridge and controlling the processing of the sample in the cartridge. Alternatively, the functionalities of the cartridge can also be integrated into the analysis device, and the analysis device can be configured to receive the sample for testing.

[0011] According to a preferred embodiment, the microfluidic system and in particular a microfluidic cartridge of the system can comprise a microfluidic architecture and reagents for performing nucleic acid amplifications, for example, one or more chambers for performing a PCR in the form of an endpoint PCR or a quantitative real-time PCR or an isothermal amplification. Furthermore, the microfluidic system and in particular the microfluidic cartridge can have an array, in particular a microarray, for hybridizing amplification products, in particular amplified nucleic acid sequences. Alternatively or additionally, the array can be a cavity array in which chemical reactions, in particular nucleic acid amplifications, can take place in the cavities formed as recesses on the surface of the array.The cavities can also contain upstream reagents, for example, primers for the respective test in the cavity. The microfluidic system, in particular the analytical device, preferably comprises a processor for executing the method according to the invention and one or more memories for storing the test results determined by the method as well as for storing a computer program for executing the method according to the invention. The invention thus also relates to a computer program comprising instructions that cause the microfluidic system, in particular the analytical device of the microfluidic system, to execute the method according to the invention, as well as to a computer-readable medium on which such a computer program is stored.

[0012] The microfluidic system, in particular the analytical device, can comprise active elements for carrying out the method, in particular actuators, for example pneumatics for actuating microfluidic pumps and valves, plungers for releasing reagents from reagent containers, heaters for carrying out the nucleic acid amplification and optics, for example a camera, for capturing the products produced by the method, in particular for detecting amplified nucleic acid sequences via luminescence, for example fluorescence.

[0013] A sample can be understood in particular as a biological sample, i.e. a sample comprising a part of a living being, in particular a body fluid or part of a body tissue, for example blood, sputum, urine or a swab.

[0014] Sample parameters refer in particular to one or more properties or characteristics of the sample. The set of sample parameters can in particular include molecular diagnostic parameters, such as the presence and / or a quantity, for example an amount or concentration, and / or other information relating to one or more pathogens, biological cells, immune entities, proteins, cell-free nucleic acids, substances and other solid, liquid or gaseous components such as blood gas values ​​in the sample. Pathogens are understood to be pathogens, particularly in the form of viruses, bacteria, fungi, worms or general single-cell organisms. Immune entities refer in particular to entities produced by a body's immune system, such as antibodies or leukocytes or parts thereof.

[0015] The input of the selection of sample parameters for which the sample is to be tested can be made, for example, via a user interface of the microfluidic system, for example via a touch-sensitive display of the system, in particular of the analysis device. Alternatively, the input can also be wireless, for example via WLAN or radio, for example via an app or online application. According to a particular embodiment, the input is made via a capture, for example an optical or wireless read-in, of information from the microfluidic cartridge used for the method. For example, the cartridge can be designed so that a user can note the selection of parameters in an area on the outside of the cartridge, for example by marking or checking off applied information on testable sample parameters, or can store the data in a readable memory of the cartridge.According to an advantageous embodiment, the one or more predetermined parameters can be selected by reading information associated with the microfluidic cartridge.

[0016] The invention advantageously provides a broadly applicable method and a broadly deployable microfluidic system that can be used for a variety of tests, in particular molecular biological tests. The invention makes it possible to select and evaluate a subset from the quantity or large group of sample parameters that can be tested by the microfluidic system, i.e., in particular, from a multiplex test. In particular, the system can be used for the respective application without hardware adaptation and with a consistent interface for the user. Using the same system for the many application cases makes it possible to scale the production of the system as needed without technical changes, thereby optimizing the use of resources. Furthermore, it is advantageously easy to add or modify testable sample parameters in the system.Any regulatory approval that may be required may also be limited to just one system. Preferably, the sample is tested not just for the selected sample parameters, but for all sample parameters of a subset of the set of sample parameters by the system. The subset preferably contains at least one non-selected sample parameter from the set of sample parameters. Preferably, the subset is a true subset of the set, i.e., it preferably contains fewer sample parameters than the set. This advantageously utilizes the fact that the system can test for multiple sample parameters in one process, preferably allowing synergy effects to be realized. In particular, the subset can contain sample parameters whose joint testing requires fewer resources overall, i.e., individual testing of these sample parameters.For example, testing involves procedural steps that are similar or even identical to tests for many of the possible sample parameters, such as sample preparation, including, for example, the separation or lysis of sample components or the amplification of sample components, in particular, the amplification of nucleic acid sequences. For example, in a test for a selected sample parameter that includes the detection and / or quantification of a specific nucleic acid sequence in the sample, other, non-selected sample parameters are also tested, which also require the detection and / or quantification of specific nucleic acid sequences, whereby the detection or quantification requires one or more identical test steps, in particular an identical or similar nucleic acid amplification.For example, the test for a selected sample parameter and one or more non-selected sample parameters can be carried out via a joint nucleic acid amplification, for example, a joint PCR or a joint isothermal amplification with multiple primers or multiple primer pairs for testing the multiple sample parameters. When performing reactions, in particular nucleic acid amplifications, in multiple wells of a well array, the subset can include sample parameters whose testing involves the use of wells located in a common area with the wells used for testing the selected sample parameters, which facilitates and accelerates subsequent processing or the optical analysis and selection of the reaction products in the wells.Depending on the selected sample parameters, a subset is preferably formed from the set, wherein the subset comprises the selected sample parameters and further non-selected sample parameters from the set, wherein the tests for the further non-selected sample parameters are carried out jointly in at least one or more method steps, in particular via a joint nucleic acid amplification. In other embodiments of the invention, the subset may additionally or alternatively also comprise predetermined sample parameters from the set that do not share any common method steps when testing for the sample parameters with other sample parameters from the subset.

[0017] The test results for the selected sample parameters can preferably be output, for example, via a display of the microfluidic system or via a particularly wireless transmission, for example via WLAN or radio, to another device, for example, a cloud architecture, a laptop, or a smartphone. Output is therefore understood in particular to mean that the test results leave the microfluidic system and are preferably communicated to a person, in particular the user who selects the selected sample parameters, and are not only stored in a memory of the microfluidic system, whereby this memory can also be located, for example, in a cloud device in a distributed architecture of the system.

[0018] Preferably, test results for non-selected sample parameters for which the sample was tested are only output after a query for these sample parameters. This has the advantage that the user initially only receives the test results for the selected sample parameters. The test results for the non-selected sample parameters can be stored in a memory of the microfluidic system or an external memory. This has the advantage that these test results can be accessed at a later time. For example, the user can later request a test for one or more of the sample parameters for the same sample for which the test results have already been stored and can thus be output without retesting; this can also be referred to as a virtual test of these already tested sample parameters.

[0019] According to a particular development of the invention, access to these test results of the tested, non-selected sample parameters can be restricted, for example, via a software setting of the microfluidic system or encryption of the test results, and preferably only after a predetermined authorization. For example, the predetermined authorization may require the entry of a code or password or the identification of the user or a third party.

[0020] In a particular embodiment of the invention, the sample parameters selectable from the set can be restricted or limited to specific sample parameters by setting the system. Changing this setting may, for example, require the entry of a code or password or the identification of the user or a third party.

[0021] Short description of the drawings

[0022] Embodiments of the invention are illustrated schematically in the drawings and explained in more detail in the following description.

[0023] It shows

[0024] Figure 1 shows an embodiment of the system according to the invention and

[0025] Figure 2 shows a flow diagram of an embodiment of the method according to the invention.

[0026] Embodiments of the invention

[0027] Figure 1 shows an embodiment of the microfluidic device according to the invention

[0028] System 1000, which is designed, for example, to detect a plurality of pathogens. The system 1000 comprises, such as the Vivalytic® platform of Robert Bosch GmbH and without fundamentally limiting the invention thereto, a cartridge 100 for receiving and processing the sample and an analysis device 200 for receiving the cartridge 100 via an opening 215 of the analysis device 200 and controlling the processing of the sample in the cartridge, as described, for example, in the documents DE 10 2016 222 075 A1 and DE 10 2016 222 072 A1.

[0029] Figure 2 shows a flowchart for an embodiment of the method 500 according to the invention, wherein the method 500 can be carried out, for example, with the system 1000 shown in Figure 1. For this purpose, the microfluidic system 1000, in particular the analysis device 200, as also described above, comprises a memory 220 and a processor 230 and a computer program stored in the system 1000, in particular a computer program stored in the memory 220, which is configured to control the system 1000, in particular the analysis device 200, for carrying out the method 500. Alternatively, the microfluidic system 1000, in particular the analysis device 200, could also be configured to be controlled via an external entity, for example, a computer infrastructure in a cloud, at least for some method steps, for example, via a WLAN or mobile radio module 240 in the analysis device 200.

[0030] In a first step 501 of the method 502, a biological sample is provided which is to be tested for the presence of pathogens and preferably their quantity in the sample, for example, a sample comprising material from a throat swab. The microfluidic system 1000 is configured to test the sample for a number of different sample parameters, in this example, for several different pathogens, for example, for the presence and quantity of influenza viruses, coronaviruses, and bacteria of different predetermined strains, for example, to determine the cause of an illness with respiratory symptoms.Possible sample parameters can be the specific identity and / or quantity of a detected strain, but also the less specific information such as whether viruses or bacteria are present in the sample in a certain minimum quantity, for example as support for an indication for the use of antibiotics.

[0031] In a second step 502 of the method, the sample parameters to be tested are selected from the set. For example, pneumonia is present, and the selected sample parameter is the presence of bacteria in a minimum amount in the sample. The selection can be entered directly by the user, for example, on the analysis device 200, for example via a touch-sensitive display 215, as indicated in Figure 1. Alternatively, the user could also use an app, for example on a smartphone, to transmit the selection to the system 1000, in particular via the WLAN or mobile radio module 240. Alternatively, a cartridge 100, into which the sample is inserted, could also be used to transfer the information about the selection to the system 1000, for example by optically reading information indicated on the cartridge 100, for example via an optical sensor of the analysis device 200.The selectable sample parameters can be restricted or limited to certain sample parameters via a system setting, so that a user must first obtain authorization to test these sample parameters, for example via the WLAN or mobile radio module 240, for example by entering a corresponding authorization, for example via the touch display 215.

[0032] In the third step 503 of the method 500, the test is carried out. In the simplest embodiment, the sample is only tested for the selected sample parameter. Preferably, however, in addition to the test for the selected sample parameter, other non-selected sample parameters are also tested, for example the specific identification of the type or strain of bacteria, for example, whether it is streptococci or staphylococci and, if applicable, which type or subspecies. For example, the microfluidic system is designed to test one or two types of viruses, in particular influenza viruses, in addition to these bacteria in the same PCR, for example because the primers to be used for the different species are coordinated in such a way that the PCR reaction products can be generated alongside one another and without interference.The subset of sample parameters considered for the test in the third step can thus include, in addition to the selected sample parameter, the additional sample parameters for distinguishing, for example, five types of bacteria and three strains of influenza viruses. In an alternative embodiment, the system can be programmed to test the samples for all other possible sample parameters in addition to the selected sample parameter. All other possible sample parameters are understood to mean, in particular, all sample parameters that can fundamentally be tested for with the system 1000 and, in particular, the cartridge 100 used, given the type of sample.

[0033] After completion of the test, in a fourth step 504, the test results are stored in a memory of the microfluidic system 1000, in particular in a memory 220 of the analysis device 200, and only the test result of the selected sample parameter is communicated to the user, for example, via a display on the system 1000, in particular on a display 215 of the analysis device 200, as indicated in Figure 1, or via transmission via the WLAN or mobile radio module 240 to another device, for example, to a smartphone via an app. According to this example, the user learns whether the bacteria are present in the sample in the minimum quantity.

[0034] If, in a fifth step 505 of the method, the user also wishes to test the sample for additional sample parameters, the test results for the additional, previously unselected sample parameters can be output without performing a new test, which can be referred to as virtual retesting. For example, the test result for the presence of bacteria may have been negative, so the user now wants to know whether influenza viruses are present in the sample. Alternatively, the test result may have been positive, and the user wants to know whether it is streptococci or staphylococci. For example, the output of these additional test results may depend on an authorization that the user must obtain, for example, via the WLAN or radio module 240 and / or by entering an authorization, in particular via the touch display 215.

Claims

Claims 1 . Method (500) for testing a sample for predetermined sample parameters using a microfluidic system (1000), in particular for detecting pathogens, wherein the microfluidic system (1000) is designed to test a sample, in particular a biological sample, for a set of different sample parameters, wherein one or more sample parameters are selected from the set, wherein the sample is tested for the selected sample parameters.

2. The method (500) according to claim 1, wherein the sample is tested for all sample parameters of a subset of the set, wherein the subset comprises the selected sample parameters and wherein the subset is in particular smaller than the set 3. The method (500) of claim 2, wherein the subset comprises at least one unselected sample parameter.

4. The method (500) of claim 2 or 3, wherein test results are output for the selected sample parameters.

5. Method (500) according to one of the preceding claims, wherein test results for non-selected sample parameters for which the sample was tested are output only after a query for these sample parameters, wherein the output of the test results for the non-selected sample parameters preferably only takes place after a predetermined release.

6. Method (500) according to one of the preceding claims, wherein the sample parameters selectable from the set can be restricted or are restricted to specific sample parameters via a setting of the system (1000).

7. The method (500) according to any one of the preceding claims, wherein the subset comprises sample parameters for the determination of which an at least partially common detection reaction, in particular a common nucleic acid amplification, for example a PCR or isothermal nucleic acid amplification, can be carried out.

8. The method (500) according to any one of the preceding claims, wherein the microfluidic system (1000) comprises a cartridge (100) in which the sample is tested.

9. Method (500) according to one of the preceding claims, wherein the microfluidic system (1000), in particular the cartridge (100), comprises a cavity array, wherein detection reactions for various sample parameters are carried out in cavities of the cavity array.

10. The method (500) according to any one of the preceding claims, wherein the one or more predetermined parameters are selected by reading information associated with the cartridge (100).

11. Method (500) according to one of the preceding claims, wherein the values ​​of the further parameters are stored / reserved for later output 12. A microfluidic system (1000) for testing a sample for predetermined sample parameters, configured to carry out a method (500) according to any one of the preceding claims. i 13. Microfluidic system (1000) according to claim 12, wherein the system (1000) comprises a cartridge (100) for receiving and processing the sample and an analysis device (200) for receiving the cartridge (100) and controlling the processing of the sample in the cartridge (100).

14. Computer program comprising instructions which cause a microfluidic system (1000) according to claim 12 or 13, in particular an analysis device (200) of the microfluidic system (1000), to carry out the method steps according to one of claims 1 to 11.

15. A computer-readable medium on which the computer program according to claim 14 is stored.

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