Method for verifying the identity of patient samples

Abstract

The present invention relates to a device for taking a patient sample, in particular for use in high-throughput sequencing analyses, in particular in next-generation sequencing (NGS) methods, comprising a container, wherein the container comprises a biological information means integrated into the device in the receiving interior of the container prior to taking samples and a container information means arranged outside of the receiving interior, wherein the container identification means provides information about the biological identification means contained in the receiving interior of the container.

Description

[0001] Labor Berlin – Charité Vivantes GmbH P70549WO METHOD FOR IDENTITY VERIFICATION OF PATIENT SAMPLES The present invention relates to a device for receiving a patient sample, in particular for use in high-throughput sequencing analyses, especially in next-generation sequencing (NGS) methods. The present invention also relates to methods for analyzing patient samples, as well as methods for verifying the identity of a patient sample. Next-generation sequencing (NGS) refers to a technology in which a large number of DNA sequences are performed in parallel. It enables the rapid analysis of a large number of samples (up to several hundred patients in a parallel analysis).However, the complex analysis protocols and the large number of samples in the multiplex process in NGS-based human genetic diagnostics can lead to unavoidable sample mix-ups, which compromise diagnostic certainty and thus pose a high risk to patients. For these reasons, the clear and reliable assignment of patient samples in this field is prone to errors and must be controlled according to risk management (e.g., according to DIN ISO 15189). One approach to reducing the risk of incorrect assignment is the automated or semi-automated tracking of sample containers using barcodes and the detection of tracking errors by recognizing sequence characteristics. One example of this is the determination of the patient's sex by checking for non-autosomal (ds gonosomal) markers, e.g., the presence of the Y chromosome in male patients.However, this method is only partially reliable, as it cannot detect mismatches in samples from patients of the same sex. The first commercially available kits (e.g., Nimagen EasySeq™ Human WES WGS Sample Tracking Kit) are available. sample-tracking-and- Twist Bioscience Human Sample ID Kit, Labor Berlin – Charité Vivantes GmbH P70549WO fragmentation-kit / Twist-Human-Sample-ID-Kit) are based on the determination of intrinsic genetic distinguishing characteristics with the character of a genetic fingerprint (individual-specific polymorphic genetic variants). For this purpose, the sample must be examined twice after DNA extraction, i.e., the determined genetic characteristics are compared between the diagnostics and the kit (see Fig. 1). For this purpose, a sample (110) is first provided. From this, the patient's DNA is sequenced, for example, as a whole exome sequencing (WES) or whole genome sequencing (WGS) diagnostic, which simultaneously determines a sample characteristic such as gender and a series of genetic polymorphisms (Guidelines for diagnostic next generation sequencing; Appendix 2: SNPs for sample identification (120). The commercial kit determines (130) and allows verification that the sample has been assigned to the correct patient. Both results (120) and (130) can be determined in parallel, for example, using a DNA sequencer (140). The results (120) and (130) are compared (150) to confirm the assignment (160). The disadvantages of the above-mentioned methods based on intrinsic characteristics are that (i) two separate measurements (120) and (130) must be performed, which in turn can lead to confusion, (ii) most commercial kits (e.g., Nimagen HEST) are only used after pre-analytical testing (DNA extraction), (iii) intrinsic characteristics are not suitable for twin / sibling diagnostics or tumor / normal tissue comparison (same genotype), and (iv) an adaptation of the diagnostics is necessary to determine the comparative characteristics. Even this complex analysis does not rule out the possibility of the primary patient sample being used as input material in the parallel analysis.The motivation of the present invention is therefore to provide a system based on extrinsic sample properties that is less prone to errors in the unambiguous and reliable assignment of patient samples by (i) measuring comparable sample properties in a sequencing process, (ii) adding extrinsic markers to the primary sample material (blood), and (iii) automatically checking sample properties. Laboratory Berlin – Charité Vivantes GmbH P70549WO The present invention provides devices and methods based on a single laboratory analysis step. By providing a device that contains a biological information medium integrated before sample collection, a control option is created that is independent of the patient sample. Such a control option is based on a sample-independent assignment between the interior of the receiving chamber and the external marking of the device (for example, bya barcode). It is therefore less prone to errors. On the other hand, the use of a biological information medium allows it to be analyzed together with the patient sample in an analytical method, for example by means of DNA sequencing. The analysis of a biological information medium added to the patient sample is inherent in sequencing (e.g. WGS or WES analysis). According to the invention, a separate test for identification determination can be dispensed with. An assignment of the recorded biological information medium to the information medium integrated in the device can take place bioinformatically. The present invention therefore relates to a device for receiving a patient sample, comprising a container with a receiving interior for receiving a patient sample, wherein the container i. a biological identification medium contained in the receiving interior of the container; and ii. an outside of the receiving interiorarranged container identification means. The biological identification means and the container identification means are integrated into the device before sampling. The container identification means provides information on the biological identification means contained in the receiving interior of the container. According to a preferred embodiment, the biological identification means can be evaluated by sequencing, particularly preferably by WGS or WES. According to a further preferred embodiment, the biological identification means is a DNA identification means, preferably a circular double-stranded DNA molecule with a DNA barcode sequence coupled to coding human DNA. Labor Berlin – Charité Vivantes GmbH P70549WO According to a further preferred embodiment, the container identification means is an optically readable identification means, preferably a barcode, particularly preferably a 1D barcode or a 2D barcode. According toAccording to a further preferred embodiment, the container identification means is an identification means that is not optically readable, preferably an electronic identification means, particularly preferably an RFID tag. According to a further preferred embodiment, the biological identification means and the container identification means are uniquely, preferably uniquely, paired with one another. According to a further preferred embodiment, the container comprises one or more than one further identification means arranged outside the receiving interior. According to a further preferred embodiment, the one or more than one further identification means is an optically readable identification means, preferably a barcode, particularly preferably a barcode selected from the list consisting of a 1D barcode and a 2D barcode. According to a further preferred embodiment, the one or more than one further identification means is selected fromthe group consisting of a physician identification means, a laboratory identification means, or a combination thereof. According to a further preferred embodiment, the physician identification means provides information about the patient from whom the patient sample originates, information about the sample collection, or a combination of two more thereof. According to a further preferred embodiment, the laboratory identification means provides laboratory-internal information. According to a further preferred embodiment, the container identification means provides information about the device. Labor Berlin – Charité Vivantes GmbH P70549WO According to a further preferred embodiment, the biological identification means comprises at least two circular double-stranded DNA molecules with a DNA barcode sequence coupled to encoding human DNA. According to a further preferred embodiment, the DNA barcode sequence of each circular double-stranded DNA molecule compriseswith a DNA barcode sequence coupled to encoding human DNA, 5 to 15 base pairs. According to a further preferred embodiment, the DNA barcode sequences of all circular double-stranded DNA molecules with a DNA barcode sequence coupled to encoding human DNA contained in the biological information medium together comprise 10 to 50 base pairs. According to a further preferred embodiment, the container is a blood tube. According to a further preferred embodiment, the container is an EDTA tube. According to a further preferred embodiment, the container contains an anticoagulant in the interior space. Particularly preferably, within the scope of the invention, a well-defined DNA sequence construct ("DNA barcode") of artificial origin serves as the biological identification means. The container identification means ("barcode") encodes the expected associated DNA sequence and thus provides a key to resolving the sequence.for analyzing a patient sample, comprising the following steps: a. Providing a device for receiving a patient sample, comprising a container with a receiving interior for receiving a patient sample, wherein the container comprises: i. a biological identification means contained in the receiving interior of the container; and ii. a container identification means arranged outside the receiving interior, Labor Berlin – Charité Vivantes GmbH P70549WO, wherein the biological identification means and the container identification means are integrated into the device before sampling, wherein the container identification means provides information about the biological identification means contained in the receiving interior of the container, and wherein the receiving interior of the container contains a patient sample; b. Mixing the biological identification means with the patient sample; c. Scanning the container identification means on the outsideof the container; d. jointly removing the patient sample and the biological identification means from the receiving space; and e. at least partially simultaneously analyzing the patient sample and the biological identification means. According to a preferred embodiment, the patient sample is a blood sample, preferably a whole blood sample, preferably EDTA blood. According to a further preferred embodiment, the container contains an anticoagulant in the receiving space. According to a further preferred embodiment, the biological identification means is a DNA identification means, preferably a circular double-stranded DNA molecule with a DNA barcode sequence coupled to encoding human DNA or a combination of two or more circular double-stranded DNA molecules with a DNA barcode sequence coupled to encoding human DNA. According to a further preferred embodiment, the at least partially simultaneously analyzing thePatient sample and the biological identification means, the sequencing of the DNA of the patient sample and the biological identification means contained in the container, preferably by means of WGS or WES. According to a further preferred embodiment, the container identification means is an optically readable identification means, preferably a barcode, particularly preferably a 1D barcode or a 2D barcode. Labor Berlin – Charité Vivantes GmbH P70549WO According to a further preferred embodiment, the container identification means is an identification means that is not optically readable, preferably an electronic identification means, particularly preferably an RFID tag. According to a further preferred embodiment, mixing the biological identification means with the patient sample comprises dissolving the biological identification means in the patient sample. According to a further preferred embodiment, the biological information means isat least partially simultaneously with or after the patient samples are placed in the receiving space of the container. Particularly preferably, in the method for analyzing a patient sample, the "DNA barcode" of the biological identification means of artificial origin identifies blood samples before pre-analysis. In the step of extracting the DNA, both the patient's DNA and the "DNA barcode" of the biological identification means are purified. In the context of the present invention, both properties are measured in parallel in one measurement, for example in diagnostics by high-throughput (NGS) sequencing. In a further aspect, the present invention relates to a method for verifying the identity of a patient sample, comprising the following steps: a) Providing a device for receiving a patient sample, comprising a container with a receiving space for receiving a patient sample, wherein the container comprises: i. abiological identification means contained in the receiving interior of the container; and ii. a container identification means arranged outside the receiving interior, wherein the biological identification means and the container identification means are integrated into the device before sampling, wherein the container identification means provides information on the biological identification means contained in the receiving interior of the container, and wherein the receiving interior of the container contains a patient sample; Labor Berlin – Charité Vivantes GmbH P70549WO b) mixing the biological identification means with the patient sample; c) scanning the container identification means on the outside of the container to identify a theoretically determined biological identification means stored in the container identification means; d) jointly removing at least a portion of the patient sample and the biologicalIdentification means from the receiving space; e) at least partially simultaneous analysis of the patient sample and the biological identification means; f) identifying the analysis results of the patient sample and the biological identification means; g) comparing the biological identification means theoretically determined on the basis of the container identification means and the analytically determined biological identification means; and h) outputting the result of the comparison. According to a preferred embodiment, the patient sample is a blood sample, preferably a whole blood sample, preferably EDTA blood. According to a further preferred embodiment, the container in the receiving space contains an anticoagulant. According to a further preferred embodiment, the biological identification means is a DNA identification means, preferably a circular double-stranded DNA molecule with a DNA barcode sequence coupled to coding human DNA or aCombination of two or more circular double-stranded DNA molecules with a DNA barcode sequence coupled to encoding human DNA. According to a further preferred embodiment, the at least partially simultaneous analysis of the patient sample and the biological identification means comprises the sequencing of the DNA of the patient sample and the biological identification means contained in the container, preferably by means of WGS or WES. According to a further preferred embodiment, the container identification means is an optically readable identification means, preferably a barcode, particularly preferably a 1D barcode or a 2D barcode. Labor Berlin – Charité Vivantes GmbH P70549WO According to a further preferred embodiment, the container identification means is an identification means that is not optically readable, preferably an electronic identification means, particularly preferably an RFID tag. According to a further preferred embodimentMixing the biological identification means with the patient sample comprises dissolving the biological identification means in the patient sample. According to a further preferred embodiment, in step f), the DNA barcode sequence is identified using exogenous marker sequences and separated from the patient DNA. According to a further preferred embodiment, the biological information means is placed in the receiving space of the container before, at least partially simultaneously with, or after the patient sample. Particularly preferably, within the scope of the method for identity verification of a patient sample, after the analysis (sequencing) of patient DNA and the DNA barcode stored in the biological information means, the DNA barcode readings are identified using exogenous marker sequences in the biological identification means and separated from the patient DNA reading in order to exclude any influence on the diagnostic quality. Subsequently, theThe measured DNA barcode sequence of the biological identification means is compared with the expected DNA sequence according to the barcode of the container identification means. If there is no match, a sample mix-up has occurred during the workflow. This avoids time-consuming and costly duplicate testing of the sample material. Furthermore, unlike the use of intrinsic genetic distinguishing features, adapting the diagnostics to these features is not necessary. A patient sample, within the scope of the present invention, is a sample that contains the patient's DNA. Labor Berlin – Charité Vivantes GmbH P70549WO Containers for patient samples are known to those skilled in the art. They are also referred to as sample vessels, vessels, or sample tubes within the scope of the present invention. Generally, commercially available sample tubes for patient samples are suitable as containers according to the present invention.A biological information medium according to the present invention comprises a biological sequence. A biological sequence in the context of the present invention is a DNA sequence, RNA sequence, DNA-RNA hybrid sequence, or amino acid sequence. The length of the biological sequence must be able to ensure the identification and differentiation of samples in high-throughput biological analysis methods. The length of the biological sequence is greater than 5. "Unique" in the context of the present invention means that each container identification means is assigned to a biological information medium. "Simultaneously" in the sense of the present invention means that the patient sample and the biological information medium are examined in one step. This must be distinguished from methods known from the prior art in which a sample must be examined twice, for example, by analyzing a biological information medium in one step andAnalyzing a patient sample in separate steps. Fig. 1 shows a method for identity verification which is known from the prior art. Fig. 2 shows a method according to the present invention for in vitro indexing. Fig. 3 shows a method according to the present invention for the identity verification of patient samples. Fig. 4 schematically shows the stations from the production of a device for identity verification according to the present invention via the sample storage up to the analysis and identity verification of patient samples. Labor Berlin – Charité Vivantes GmbH P70549WO Fig. 5 shows a device according to the present invention. Fig. 6 shows sequenced PUC19 vectors with ERH / GAPDH sequence with and without barcode in the plasmid. The present invention relates to a device for receiving a patient sample comprising a container with an interior for receiving a patient sample, wherein the container i.a biological identification means contained in the receiving interior of the container; and ii. a container identification means arranged outside the receiving interior. The biological identification means and the container identification means are integrated into the device before sampling. The container identification means provides information about the biological identification means contained in the receiving interior of the container. A patient sample within the meaning of the invention is a sample that contains the patient's DNA. Examples include blood, hair, saliva, urine, and skin. Preferably, according to the present invention, the patient sample is a blood sample. Preferred blood samples are selected from the list consisting of whole blood, dried blood, and blood cells. Particularly preferred patient samples are whole blood samples. Preferably, the blood sample is stabilized by an anticoagulant, e.g., EDTA. Containers for such patient samples are known.Within the scope of the present invention, they are also referred to as sample vessels, vessels, or sample tubes. In principle, within the scope of the present invention, commercially available sample vessels or sample tubes for such patient samples are suitable as containers. Preferred containers within the scope of the present invention are blood tubes. Preferred blood tubes are EDTA tubes. The containers preferably have a temporary closure. Temporary closures can be, for example, lids or, in the case of Monovettes, a membrane screw cap. Within the scope of the present invention, the closure of the container is understood as part of the container. Labor Berlin – Charité Vivantes GmbH P70549WO The container can have a volume of 0.5 mL to 15 mL, for example, 1 mL to 5 mL, preferably 1.5 mL to 2.5 mL. The biological information medium is preferably evaluable by sequencing, particularly preferably by WGS or WES. It is preferably a DNA information medium, which within the scope ofof the present invention means that the biological information means preferably comprises a DNA sequence. Preferably, circular double-stranded DNA molecules with a DNA barcode sequence coupled to coding human DNA are used. In the following, "circular double-stranded DNA molecules with a DNA barcode sequence" are also referred to as "circular DNA molecules." The coding human DNA sequence ensures that the biological identification means is taken into account during DNA sequencing of the sample. Preferably, the coding human DNA sequence has no pathological significance, so that the diagnosis is not falsified. The DNA barcode sequence is coupled to the coding human DNA sequence and is therefore sequenced during DNA sequencing of the sample. Basically, all plasmids (except human mitochondria because of diagnostic relevance) that can carry a plasmid DNA barcode are suitable as circular DNA molecules, provided theyare linked to a section of human coding DNA sequence. Examples of such plasmids are available via the National Library of Medicine database at https: / / www.ncbi.nlm.nih.gov / nuccore / A plasmid DNA barcode is a circular vector construct. Such circular vector constructs can be selected from the list consisting of pBR322, pBluescriptII and pUC plasmids such as pUC18 and pUC19. Within the scope of the present invention, it may also be preferred for the biological identification means to comprise at least two circular double-stranded DNA molecules with a DNA barcode sequence linked to coding human DNA. It is also possible for the biological identification means to comprise more than two circular double-stranded DNA molecules with a DNA barcode sequence linked to coding human DNA, for example 3 to 10 plasmids. In principle, a combination of more than 10 plasmids is also possible within the scope of the presentInvention possible. Laboratory Berlin - Charité Vivantes GmbH P70549WO The DNA sequence or DNA barcode sequence can in principle have any length. Preferably, the DNA sequence or DNA barcode sequence is a sequence with a length of at least 5 base pairs (bp), for example of at least 6 bp or of at least 7 bp or of at least 8 bp or of at least 9 bp, particularly preferably of at least 10 bp. Preferably, the DNA sequence or DNA barcode sequence is a sequence with a length of not more than 50 base pairs (bp), for example of not more than 40 bp or of not more than 30 bp or of not more than 20 bp, particularly preferably of not more than 15 bp. The DNA barcode sequence of each circular double-stranded DNA molecule with a DNA barcode sequence coupled to coding human DNA preferably comprises 5 to 15 base pairs. Preferably, the DNA barcode sequences comprise all circular double-stranded DNA molecules with a DNA barcode sequence coupled tocoding human DNA contained in the biological information medium, together 10 to 50 base pairs. The biological information medium is preferably evaluable by means of biological or biochemical sample analysis. Such analysis preferably comprises the analysis of the biological sequence contained in the container, e.g., the DNA, the RNA, or the amino acid sequence of the biological information medium. Preferred DNA analysis methods are high-throughput sequencing, in particular WES and WGS analyses. Preferred RNA and amino acid sequence analysis methods are RNA and protein sequencing methods. Such RNA and protein sequencing methods are well known to the person skilled in the art. The container identification means provides information about the biological identification means contained in the interior of the container. In some cases, the container identification means may be a tangible label (e.g., printed, drawn, or onattached to the vessel or another object connected to the vessel). In some cases, the container identification means may be an intangible label (e.g., digitally rendered or stored as metadata). Labor Berlin – Charité Vivantes GmbH P70549WO The container identification means may be an optically readable identification means, for example a code, a mark, an identification number, or the like. Examples of suitable codes are one-dimensional (1D) codes, two-dimensional (2D) codes, three-dimensional (3D) codes, Quick Response (QR) codes, barcodes, encrypted codes, or alphanumeric codes. The container identification means may be a non-optically readable identification means, for example, an acoustic code, an electronic code, or a haptic code. Preferred non-optically readable identification means are electronic identification means, particularly preferably RFID tags. TheContainer identification means can be read for linking to the biological identification means. For example, a container identification means, in particular an optical container identification means, can be scanned with a scanner or other optical detector. The scanner or other optical detector can be communicatively connected to a user device (e.g., a computer, mobile phone, laptop, tablet, or other portable device). The container identification means, in particular a non-optical container identification means, can be detected with another type of sensor (e.g., a microphone or other acoustic sensor) or detector, including a non-optical detector. In some cases, further information stored in the container identification means can be accessed via a hyperlink. The container identification means is preferably a barcode. The barcode is particularly preferably a1D barcode or a 2D barcode. The 2D barcode is preferably a QR code. The container identification means is arranged outside the receiving space of the container. The container identification means is preferably arranged on the outside of the container. It can in principle be located anywhere on the outside of the container, for example on the outer wall or on the bottom. The container identification means should be arranged so that it can be read from the outside. It is also possible for an additional container identification means to be located on the closure of the container. In this case, the container identification means Labor Berlin – Charité Vivantes GmbH P70549WO outside the receiving space of the container and the container identification means on the closure of the container are identical. Preferably, the biological information means and the container identification means are uniquely paired with each other. The pairing can, for example, already be achieved by theManufacturer. In this case, the device is already provided with the biological identification means by the manufacturer. The container can optionally comprise one or more further identification means arranged outside the receiving interior. The one or more further identification means can in principle be in the same form as the container identification means. Preferably, the one or more further identification means is an optically readable identification means, preferably a barcode, particularly preferably a barcode selected from the list consisting of a 1D barcode and a 2D barcode. The one or more further identification means can also be an identification means that is not optically readable, preferably an electronic identification means, particularly preferably an RFID tag. Further identification means are, for example, doctor identification means, i.e.Person (doctor, practice, other institution) applied identification means. These can contain, for example, information about the patient, the person taking the sample and the sample collection. Other identification means are, for example, laboratory identification means, i.e. identification means applied by the sample analyzing laboratory. These can contain, for example, information for the laboratory-internal assignment of patient characteristics and / or request characteristics (e.g. order number, order entry identifier, etc.). The biological information medium is contained in the interior of the container. The biological information medium can be contained in solid or liquid form in the interior of the container. Labor Berlin – Charité Vivantes GmbH P70549WO In solid form, the biological information medium can be contained both as a pure substance and in combination with other components, for example a pH stabilizer (slightly basic). IsContaining the biological information agent in liquid form is understood within the scope of this invention to mean that it is dissolved or suspended in a liquid or in a combination of two or more liquids. Preferred liquids are H2O or TE buffer, i.e., a buffer consisting of aqueous solutions of TRIS and EDTA. It is also possible for the biological information agent to be bound to the inner wall of the container prior to the addition of the patient sample and / or other components, or to be present in lyophilized form in the receiving space. In this embodiment, the biological information agent is detached from the inner wall of the receiving space by adding the patient sample and / or other components. The container may contain further components in addition to the biological information agent. Further components within the scope of the present invention are - anticoagulant, preferably EDTA, - stabilizers, - H2O, and - buffer. Preferably, it containsthe container in the receiving interior contains an anticoagulant. Preferably, the container in the receiving interior contains EDTA as an anticoagulant. FIG. 5 shows an example of a device (500) according to the invention for receiving a patient sample. The device has a container (510) with a receiving interior (520). A biological identification means (530) is contained in the receiving interior. A container identification means (540) in the form of a barcode is applied outside the receiving interior on the outside of the container. The container is closed with a lid (550). Labor Berlin – Charité Vivantes GmbH P70549WO In a further aspect, the present invention relates to a method for analyzing a patient sample, comprising the following steps: a. Providing a device according to the invention for receiving a patient sample, wherein the receiving interior of the container contains a patient sample; b. Mixing the biologicalIdentification means with the patient sample; c. Scanning the container identification means on the outside of the container to identify a theoretically determined biological identification means stored in the container identification means; d. Jointly removing at least a portion of the patient sample and the biological identification means from the receiving interior; and e. At least partially simultaneously analyzing the patient sample and the biological identification means. The device provided according to step a. corresponds to a device for receiving a patient sample described above. The exemplary and preferred embodiments described above are also exemplary or preferred for the device for receiving a patient sample provided in the method according to the invention. According to step a. of the method for analyzing a patient sample, the receiving interior of the container contains aPatient sample. To provide the device according to the invention for receiving a patient sample, wherein the receiving interior of the container contains a patient sample, the biological information medium can be introduced into the receiving interior of the container before, at least partially simultaneously with, or after the patient sample. For example, a practice can use a device according to the invention that includes the biological information medium by the manufacturer. It is also possible for a practice to provide a device according to the invention by itself supplying the biological information medium to the receiving interior of the container. In this case, the biological information medium can be supplied to the receiving interior of the container before, at least partially simultaneously with, or after the patient sample. It is also possible for a laboratory commissioned with sample analysis to provide a device according to the invention by itselfthe biological laboratory Berlin – Charité Vivantes GmbH P70549WO supplies the information medium to the receiving space of the container. In this case, the biological information medium is supplied to the receiving space of the container after the patient sample. The patient sample and the biological identification means are mixed according to step b. of the method for analyzing a patient sample. They are present in the receiving space of the container as a mixture. The mixture can be homogeneous or heterogeneous. Preferably, the biological identification means is present in dissolved form in the receiving space of the container. Preferably, the biological identification means is dissolved in the patient sample. The storage of the device for receiving a patient sample containing a patient sample in the receiving space of the container can lead to a phase separation of the patient sample. Therefore, within the scope of the method according to the invention, it is preferred, before removing at least part of thePatient sample, a suitable mixing process must be carried out. Suitable mixing processes are well known to the person skilled in the art. Examples of such mixing processes are shaking, stirring and swirling. Carrying out a mixing process before taking at least a portion of the patient sample also ensures better mixing of the biological identification agent with the patient sample. Preferably, such a mixing process is carried out no more than 5 minutes, for example no more than 4 minutes or no more than 3 minutes or no more than 2 minutes, particularly preferably no more than 1 minute before the joint removal of at least a portion of the patient sample and the biological identification agent from the receiving space of the container. If the sample is homogeneous even without carrying out a mixing process, mixing is preferably dispensed with. According to step c. of the method forWhen analyzing a patient sample, the container identification means on the outside of the container is scanned to identify a theoretically determined biological identification means stored in the container laboratory Berlin – Charité Vivantes GmbH P70549WO identification means. Optionally, further identification means on the outside of the container can also be scanned. For example, the container identification means can be scanned with a scanner or other optical detector. The scanner or other optical detector can be communicatively connected to a user device (e.g., a computer, mobile phone, laptop, tablet, or other portable device). Scanning the container identification means can lead the user to a database in which pairs of container and biological information means are stored. According to step d. of the method for analyzing a patient sample, at least onePart of the patient sample and the biological information medium are taken together from the interior of the container. The volume of the taken sample containing at least part of the patient sample and the biological information medium is preferably in the range from 1 μL to 1 mL, for example from 10 to 500 μL, particularly preferably from 40 to 200 μL. According to step e. of the method for analyzing a patient sample, the patient sample and the biological information medium are analyzed at least partially simultaneously. The at least partially simultaneous analysis of the patient sample and the biological information medium preferably comprises the sequencing of the DNA of the patient sample and the biological information medium contained in the container. Exome sequencing (WES) or genome sequencing (WGS) is particularly preferably carried out. DNA sequencing is particularly preferably carried out in accordance with DIN EN ISO 15189. These methods allowthat in one analysis step, all DNA of a sample, for example the DNA of a patient and the DNA of the biological information medium also present in the sample, is sequenced. This method for assigning a patient sample to a biological identification medium is shown as an example in FIG. 2. A plasmid with a DNA barcode coupled to coding human DNA (210) is used as the biological information medium. It is located in the receiving space of a blood tube (220). Labor Berlin – Charité Vivantes GmbH P70549WO The blood tube has a 1D barcode on the outside, which is scanned before analysis. It is located in the receiving space of a blood tube, which also contains a blood sample from a patient (230). The container identification means, which is located on the outside of the blood tube, is scanned (not shown). Subsequently, a portion of the patient sample and the biological identification means are extracted from theThe patient's DNA contained in the blood and the biological information means are analyzed at least partially simultaneously (250) after the joint removal (240). In a further aspect, the present invention relates to a method for identity verification / sample tracking of a patient sample, comprising the following steps: a) providing a device according to the invention for receiving a patient sample, wherein the receiving interior of the container contains a patient sample; b) mixing the biological identification means with the patient sample; c) scanning the container identification means on the outside of the container to identify a theoretically determined biological identification means stored in the container identification means; d) jointly removing at least part of the patient sample and the biological identification means from theReceiving interior; e) at least partially simultaneous analysis of the patient sample and the biological identification means; f) identifying the analysis results of the patient sample and the biological identification means; g) comparing the biological identification means theoretically determined on the basis of the container identification means and the analytically determined biological identification means; and h) outputting the result of the comparison of the container identification means and the biological identification means. The device provided according to step a) corresponds to a device for receiving a patient sample described above. The exemplary and preferred embodiments of the device and steps a. to e. of the method for analyzing a patient sample described above are also exemplary or preferred for the device provided in the method according to the invention.Device for receiving a patient sample. According to step a), a device for receiving a patient sample is provided. The receiving interior of the container of the device contains a patient sample. The patient sample and the biological identification means are mixed according to step b) of the method for analyzing a patient sample. According to step c) of the method for verifying the identity of a patient sample, the container identification means on the outside of the container is scanned. By scanning the container identification means on the outside of the container, the biological identification means present in the receiving interior of the container of the device can be identified. It is stored in the container identification means. The biological identification means thus identified is hereinafter referred to as the “theoretically determined biological identification means.” According to step d) of the method forFor the identity verification of a patient sample, at least a portion of the patient sample and the biological identification means are removed together from the receiving chamber. According to step e) of the method for the identity verification of a patient sample, the patient sample and the biological information means are analyzed at least partially simultaneously. Steps b) to e) correspond to the steps b) to e) described above for analyzing a patient sample. The exemplary and preferred embodiments of steps b) to e) described above are also exemplary or preferred for the steps c) to e) carried out in the method according to the invention for the identity verification of a patient sample. According to step f) of the method for the identity verification of a patient sample, the analysis results of the patient sample and the biological identification means are identified. After the analysis (in particular sequencing) ofPatient DNA and the DNA barcode stored in the biological information medium, the DNA barcode readings can be identified using exogenous marker sequences in the biological identification medium and separated from the patient DNA reading. This allows the DNA barcode to be used for subsequent identity verification of the patient sample. The biological identification medium identified in this way is hereinafter referred to as the "analytically determined biological identification medium." Even before the data is processed for the purpose of molecular diagnostics, the barcode-containing readings are bioinformatically filtered out of the data set, and the data is subsequently analyzed for diagnostic purposes. This can also prevent the analyzed DNA of the biological identification medium from influencing the diagnostic quality of the analysis result of the patient sample. According to step g) of theIn a method for identity verification of a patient sample, the theoretically determined biological identification means based on the container identification means and the analytically determined biological identification means are compared. If the theoretically determined biological identification means and the analytically determined biological identification means are identical, the identity verification of a patient sample is positive. If the theoretically determined biological identification means and the analytically determined biological identification means are not identical, the identity verification of a patient sample is negative. A method for identity verification of a patient sample is shown as an example in FIG. 3. A plasmid with a DNA barcode (310) is used as the biological information means. It is located in the interior of a blood tube (320). The blood tube has a 1D barcode on the outside, whichis scanned before analysis. The receiving interior of the blood tube also contains a patient's blood sample (330). The container identification means located on the outside of the blood tube is scanned (not shown) to determine which biological identification means is located in the receiving interior (theoretically determined biological identification means). Subsequently, a portion of the patient sample and the biological identification means are removed from the receiving interior of the blood tube (340). The patient's DNA from the blood and the biological information means are analyzed at least partially simultaneously (350) after the joint removal (340). The analysis results of the patient sample and the biological identification means are identified (360) by comparing the DNA barcode sequence with the DNA sequence inherent in the biological identification means.Sequence properties are identified and separated from the patient's DNA, so that an analytically determined biological identification means is identified. The theoretically determined biological identification means and the analytically determined biological identification means are compared (370), and the result of the comparison is output bioinformatically (390). FIG. 4 shows a further exemplary embodiment of the present invention. A device according to the invention is provided by a manufacturer 410. The device is a blood tube with a 1D barcode on the outside. It contains two unique, distinguishable plasmids with DNA barcodes (410, 412) and an anticoagulant (for example, EDTA). The device according to the invention is delivered by the manufacturer to a medical practice or hospital (420). There, the 1D barcode on the outside of the blood tube is scanned and assigned to a patient. TheBlood is drawn from the patient and collected in the scanned blood tube. For further analysis, the blood tube is forwarded to a laboratory (430), which scans the 1D barcode on the outside of the blood tube. As described above, the blood and the plasmid are simultaneously sequenced and identified. At the end of the analysis, the identity of the patient sample can be verified based on a comparison of the theoretically determined biological identifier and the analytically determined biological information agent.

[0002] Labor Berlin – Charité Vivantes GmbH P70549WO Examples Biological information medium A vector PUC19 coupled to human coding DNA and a barcode sequence of 10 base pairs in length is used as the biological information medium. The PUC19 vector with DNA barcode and ERH / GAPDH sequence (IDT Technologies) has the following sequence: TCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCCTAGACGGTCACA GCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTG GCGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCA AATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCCATTCGC CATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCATCGCTATTACGCCA GCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAG TCACGACGTTGTAAAACGACGGCCAGTGCAACGCGATGACGATGGATAGCGATTCATCGATG AGCTGACCCGATCGCCGCCGCCGGAGGGTTGCGTTTGAGACAGGCGACAGAT DNA-BC TTTGCTGGTACAGCCTACCAAGAGGCCAGAAGGCAGAACTTATGCTGACTACGAATCTGTGA ACAGTCCCTCTATCACATATGACATCAGTCAGTTGTTTGATTTCATCGATGATCTGGCAGACCT C DNA-BC GGCACCGTCAAGGCTGAGAACGGGAAGCTTGTCATCAATGGAAATCCCATCACCATCTTCCA GGA DNA-BC ATCAGTTCTGGACCAGCGAGCTGTGCTGCGACTCGTGGCGTAATCATGGTCATAGCTGTTTC CTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTA AAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCT TTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAG GCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTT CGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAG GGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAA GGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGAC GCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGTCGCTTACCGGATACCTGTCCGCCTTTCT CCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGG TCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTT ATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGG TGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTT ACCTTCGGAAAAAAGTTGGTAGCTCTTTGATCCGGCAAACAAACCAGAGGTGGCTGGCTGCTGAAGCCAGTT TTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATC TTTTCTACGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGA TTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAATGAAGTTAATTAAATCAATCTAATTAATTAG THE COGECTIONSATIONGIAGESTGETGAGETGAGETGETGETGETGETGETGETGETGESTHERGICTIONS THE MEMGER METHIGE MECTUCTIONS ALLY MECTIONS GGAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGCTGECTGETGETGETGETGETGENCGCTGENCGCTGESLICGCTCGCTCGGCTCGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGE – Charité Vivantes GmbH P70549WO CCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAA CTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGTAAGTAGTTCGCCAG TTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGTCGTTCCGTTCGGTTCGTTGCG GTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGT GCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCT TTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTT GCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTC ATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGT TCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTG GGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATG TTGAATACTCATACTCTACCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGA GCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCC GAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCG TATCACGAGGCCCTTTCATC In the sequence above, human DNA sequences are indicated in italics. Instead of the respective DNA barcode sequences themselves, "DNA-BC" is inserted as a placeholder (underlined and in bold). DNA-BC barcode sequence DNA-BC1 GTCTTCCGAC DNA-BC2 GAATGACTCC DNA-BC3 CCTGATCGTA DNA-BC4 AGAGATGCGT DNA-BC5 CGGTACACAT DNA-BC6 CTGTTCCACA DNA-BC7 AGTGGACTCT DNA-BC8 CGCGTATCATDNA-BC9 AAGAAGCCAT DNA-BC10 TATGCACCAC An example of a sequenced PUC19 vector with ERH / GAPDH sequence without barcode in the plasmid (610) and a sequenced PUC19 vector with ERH / GAPDH sequence and DNA barcode (DNA-BC1) in the plasmid (620) are shown in Figure 6. Preparing the blood sample: 1 mL of a whole blood sample is prepared in an EDTA blood tube (BD Vacutainer). Laboratory Berlin – Charité Vivantes GmbH P70549WO 0.05 ng of the above-described PUC19 vector with DNA barcode (DNA-BC1) and ERH / GAPDH sequence (IDT Technologies) is added to the blood sample. The blood tube is labeled with a QR code on the outside. The QR code encodes the DNA barcode sequence (DNA-BC1). Sample analysis - A practice receives an EDTA sample tube with a biological identifier and QR code from the manufacturer. - The practice draws blood from the patient and sends the filled blood tube to an external laboratory for analysis. - The external laboratory scans the QR code andsequences the DNA contained in the blood tube, i.e., the PUC19 vector with DNA barcode (DNA-BC1) and ERH / GAPDH sequence, as well as the patient DNA, using WES / WGS according to DIN EN ISO 15189. As part of quality assurance, the external laboratory is required to take measures to minimize the risk of sample mix-ups: - All plasmid barcode readings from the sequencing library are extracted in silico. - The sequenced (analytically determined) barcode is compared with the scanned (theoretically determined) barcode and checked for agreement with the expected barcode sequence(s). - If the external laboratory discovers an incorrect assignment, it informs the sending practice, for example, via a digital communication channel. - The sequencing library is cleaned of plasmid barcodes to avoid influencing the diagnostic results. - The plasmid barcode-cleaned sequencing library is subjected to genetic diagnostic testing.

Claims

Labor Berlin – Charité Vivantes GmbH P70549WO Claims 1. A device for receiving a patient sample, comprising a container with a receiving interior for receiving a patient sample, wherein the container comprises: i. a biological identification means contained in the receiving interior of the container; and ii. a container identification means arranged outside the receiving interior, wherein the biological identification means and the container identification means are integrated into the device before sampling, and wherein the container identification means provides information about the biological identification means contained in the receiving interior of the container.

2. The device according to claim 1, wherein the biological identification means is analyzable by sequencing, particularly preferably by WGS or WES. 3.The device according to one of the preceding claims, wherein the biological identification means is a DNA identification means, preferably a circular double-stranded DNA molecule with a DNA barcode sequence coupled to encoding human DNA.

4. The device according to one of the preceding claims, wherein the container identification means is an optically readable identification means, preferably a barcode, particularly preferably a 1D barcode or a 2D barcode.

5. The device according to one of claims 1 to 3, wherein the container identification means is an identification means that is not optically readable, preferably an electronic identification means, particularly preferably an RFID tag.

6. The device according to one of the preceding claims, wherein the biological identification means and the container identification means are uniquely, preferably uniquely, paired with one another. Labor Berlin – Charité Vivantes GmbH P70549WO 7. The device according to one of the preceding claims, wherein the container comprises one or more further identification means arranged outside the receiving interior.

8. The device according to claim 7, wherein the one or more further identification means is an optically readable identification means, preferably a barcode, particularly preferably a barcode selected from the list consisting of a 1D barcode and a 2D barcode.

9. The device according to one of claims 7 or 8, wherein the one or more further identification means is selected from the group consisting of - a doctor identification means, - a laboratory identification means, or - a combination thereof. 10.The device of claim 9, wherein the physician identification means provides information about the patient from whom the patient sample was obtained, information about the sample collection, or a combination of two or more thereof.

11. The device of claim 9, wherein the laboratory identification means provides laboratory-internal information.

12. The device of any one of the preceding claims, wherein the container identification means provides information about the device.

13. The device of any one of the preceding claims, wherein the biological identification means comprises at least two circular double-stranded DNA molecules having a DNA barcode sequence coupled to encoding human DNA.

14. The device of any one of the preceding claims, wherein the DNA barcode sequence of each circular double-stranded DNA molecule having a DNA barcode sequence coupled to encoding human DNA comprises 5 to 15 base pairs. Labor Berlin – Charité Vivantes GmbH P70549WO 15. The device according to one of the preceding claims, wherein the DNA barcode sequences of all circular double-stranded DNA molecules with a DNA barcode sequence coupled to coding human DNA contained in the biological information medium together comprise 10 to 50 base pairs.

16. The device according to one of the preceding claims, wherein the container is a blood tube.

17. The device according to claim 16, wherein the container is an EDTA tube.

18. The device according to one of the preceding claims, wherein the container contains an anticoagulant in the receiving interior.

19. A method for analyzing a patient sample comprising the following steps: a. Providing a device for receiving a patient sample comprising a container with a receiving interior for receiving a patient sample, the container comprising: i.a biological identification means contained in the receiving interior of the container; and ii. a container identification means arranged outside the receiving interior, wherein the biological identification means and the container identification means are integrated into the device prior to sampling, wherein the container identification means provides information about the biological identification means contained in the receiving interior of the container, and wherein the receiving interior of the container contains a patient sample; b. mixing the biological identification means with the patient sample; c. scanning the container identification means on the outside of the container to identify a theoretically determined biological identification means stored in the container identification means; d.jointly removing at least a portion of the patient sample and the biological identification means from the receiving chamber; and Labor Berlin – Charité Vivantes GmbH P70549WO e. at least partially simultaneous analysis of the patient sample and the biological identification means.

20. A method for verifying the identity of a patient sample, comprising the following steps: a) providing a device for receiving a patient sample, comprising a container with a receiving interior for receiving a patient sample, wherein the container comprises: i. a biological identification means contained in the receiving interior of the container; and ii. a container identification means arranged outside the receiving interior, wherein the biological identification means and the container identification means are integrated into the device before sampling, wherein the container identification means provides information about the biological identification means contained in the receiving interior of the container, and wherein the receiving interior of the container contains a patient sample;b) Mixing the biological identification means with the patient sample; c) Scanning the container identification means on the outside of the container to identify a theoretically determined biological identification means stored in the container identification means; d) Jointly removing at least a portion of the patient sample and the biological identification means from the receiving space; e) At least partially simultaneously analyzing the patient sample and the biological identification means; f) Identifying the analysis results of the patient sample and the biological identification means; g) Comparing the theoretically determined biological identification means based on the container identification means and the analytically determined biological identification means;and h) outputting the result of the comparison.

21. The method according to one of claims 19 or 20, wherein the patient sample is a blood sample, preferably a whole blood sample, preferably EDTA blood. Labor Berlin – Charité Vivantes GmbH P70549WO 22. The method according to any one of claims 19 to 21, wherein the container contains an anticoagulant in the receiving interior.

23. The method according to any one of claims 19 to 22, wherein the biological identification agent is a DNA identification agent, preferably a circular double-stranded DNA molecule with a DNA barcode sequence coupled to encoding human DNA or a combination of two or more circular double-stranded DNA molecules with a DNA barcode sequence coupled to encoding human DNA.

24. The method according to any one of claims 19 to 23, wherein the at least partially simultaneous analysis of the patient sample and the biological identification agent comprises sequencing the DNA of the patient sample and the biological identification agent contained in the container, preferably by means of WGS or WES. 25.The method according to any one of claims 20 to 24, wherein the container identification means is an optically readable identification means, preferably a barcode, more preferably a 1D barcode or a 2D barcode.

26. The method according to any one of claims 19 to 25, wherein the container identification means is an identification means that is not optically readable, preferably an electronic identification means, more preferably an RFID tag.

27. The method according to any one of claims 19 to 26, wherein mixing the biological identification means with the patient sample comprises dissolving the biological identification means in the patient sample.

28. The method according to any one of claims 20 to 27, wherein in step f) the DNA barcode sequence is identified using exogenous marker sequences and separated from the patient DNA. Labor Berlin – Charité Vivantes GmbH P70549WO 29. The method according to any one of claims 19 to 28, wherein the biological information medium is added to the receiving interior of the container before, at least partially simultaneously with or after the patient sample.

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