Method and apparatus for hemolysis detection

Abstract

A sensor assembly enabling hemolysis detection with a diagnostic analyzer. The sensor assembly may be included in a diagnostic cartridge, which may be coupled to a diagnostic analyzer, and to which a sample fluid may be delivered. The sensor assembly includes: a primary oxygen sensor configured to provide a first measurement; and modifying an oxygen sensor comprising an oxidizing agent configured to oxidize iron of hemoglobin to methemoglobin, and configured to provide a second measurement. A hemolysis detection module of the diagnostic analyzer provides a level of hemolysis in the sample based on a difference between the first measurement and the second measurement. Numerous other diagnostic analyzers, diagnostic kits, and detection methods are also provided.

Description

[0001] This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Application No. 63 / 482,123, filed January 30, 2023. The entire contents of the above patent application are hereby expressly incorporated herein by reference. Technical Field

[0002] The present application relates to diagnostic testing methods and apparatus, and more particularly to methods and apparatus that enable detection of hemolysis levels in a blood sample. Background Art

[0003] Hemolysis is the destruction of red blood cells, which in some cases can result from phlebotomy or pre-analytical reasons associated with sample handling. For example, hemolysis of a blood sample can be caused by the use of an incorrect needle size, improper tube mixing, incorrect tube filling, excessive use of aspiration, prolonged tourniquet application, prolonged storage, extreme temperatures, delayed processing, and / or other difficulties in blood sample collection or sample handling.

[0004] Similarly, hemolysis in a blood sample may occur due to certain diseases or medical conditions, such as hemolytic anemia, autoimmune diseases, bone marrow failure, and inherited blood disorders such as sickle cell disease or thalassemia.

[0005] Traditionally, hemolysis is detected by manual visual inspection of a blood sample by a technician after separating the plasma (or serum) fraction (i.e., by centrifugation) and comparing the plasma (or serum) color to a color hemolysis chart. This chart shows the color of the separated sample as it correlates with increasing concentrations of free (extracellular) hemoglobin contained in the plasma (or serum). Thus, a technician can determine the hemolysis index based on the visual color of the separated plasma (or serum).

[0006] Hemolysis, caused by improper or inappropriate specimen collection procedures, is a highly undesirable condition that can impact the accuracy and reliability of blood gas test results. For example, the impact of in vitro hemolysis on measured potassium concentrations is well known. In such cases, the reported potassium concentration may be clinically inaccurate, with the magnitude of the inaccuracy depending on the extent of hemolysis. Many other analytes can be affected by the biological and analytical interference effects of in vitro hemolysis. For example, the presence of free (extracellular) hemoglobin in a blood sample can have properties that can interfere with certain types of diagnostic tests, such as those using optical measurement techniques. In particular, the presence of free extracellular hemoglobin in a sample can interfere with certain assays due to its absorption properties. Therefore, for laboratory or bedside testing, identifying samples containing hemolysis is desirable so that certain results can be flagged as potentially suspicious and / or redrawn. Furthermore, automated testing is desirable to minimize the subjectivity of manual visual hemolysis determinations by medical personnel and to expedite the hemolysis detection process. Eliminating the centrifugation process for hemolysis testing would also be beneficial.

[0007] Therefore, apparatus and methods are desired that enable automated hemolysis detection and increased speed, and that minimize or eliminate the judgment and inconsistencies associated with manual visual inspection. Summary of the Invention

[0008] In some embodiments provided herein, a diagnostic sensor assembly is provided. The diagnostic sensor assembly may be implemented in a diagnostic cartridge configured to be connected to a diagnostic analyzer to determine the level of hemolysis in a sample. The diagnostic sensor assembly includes: a sample inlet configured to receive a sample; a sample path extending from the sample inlet; a primary oxygen sensor configured to contact the sample along the sample path; and a modified oxygen sensor configured to contact the sample along the sample path, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin (Fe 2+ to Fe 3+ ), i.e., changing the configuration of its heme iron from the ferrous state to the ferric state. Based on the signals obtained from the primary oxygen sensor and the modified oxygen sensor, the quantitative level of hemolysis in the sample can be obtained.

[0009] In some embodiments provided herein, a diagnostic analyzer includes: an analyzer body including a cartridge receiver and an electrical connector; a diagnostic cartridge receivable in the cartridge receiver, the diagnostic cartridge including: a cartridge body configured to couple to the cartridge receiver, the cartridge body including: electrical contacts capable of coupling to the electrical connector when the cartridge body is received in the cartridge receiver; a sample inlet configured to receive a sample; a sample passage extending from the sample inlet into the cartridge body and configured to receive the sample therein; a primary oxygen sensor configured to contact the sample in the sample passage and to provide a first measurement of the sample; a modified oxygen sensor configured to contact the sample in the sample passage, the modified oxygen sensor including a sensor configured to oxidize hemoglobin to methemoglobin (Fe 2+ to Fe 3+ ) and is configured to provide a second measurement result of the sample; and a controller coupled to the electrical connector, the controller being configured to receive the first measurement result and the second measurement result of the sample and being configured to provide a hemolysis level in the sample based on the first measurement result and the second measurement result.

[0010] In some embodiments provided herein, a method for determining hemolysis of a sample is provided. The method includes coupling a diagnostic cartridge to a cartridge receiver of a diagnostic analyzer, the diagnostic cartridge including a sample inlet configured to receive a sample, a sample passage extending from the sample inlet into a cartridge body, a primary oxygen sensor configured to contact the sample in the sample passage, a modified oxygen sensor configured to contact the sample in the sample passage, the modified oxygen sensor including a sensor configured to oxidize hemoglobin to methemoglobin (Fe 2+ to Fe 3+ ) an oxidant; passing the sample through the sample passage and into contact with the primary oxygen sensor and the modified oxygen sensor; obtaining a first measurement of the sample from the primary oxygen sensor; obtaining a second measurement of the sample from the modified oxygen sensor; and providing a level of hemolysis in the sample based on the first and second measurements.

[0011] Other features and aspects of the disclosure will become more apparent from the following detailed description, claims, and accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The drawings described below are for illustrative purposes only and are not necessarily drawn to scale. The drawings are not intended to limit the scope of the present disclosure in any way.

[0013] Figure 1Illustrated is a perspective view of one example of a diagnostic analyzer (eg, for use in a point-of-care location) including a diagnostic cartridge (including a diagnostic sensor assembly) receivable in a cartridge receptacle according to embodiments provided herein.

[0014] Figure 2 Illustrated is a bottom plan view of one example of a diagnostic cartridge including a sensor assembly configured to provide measurements for use in detecting hemolysis levels according to embodiments provided herein.

[0015] Figure 3A The diagram shows the Figure 2 FIG2 is a partial cross-sectional view of an example of a diagnostic cartridge taken along section line 3A-3A of FIG2 and illustrates an exemplary configuration of a sensor assembly in which the oxidant is contained in an outer membrane layer covering an inner electrolyte chamber. According to embodiments provided herein, the membrane layer can be homogeneous or heterogeneous.

[0016] Figure 3B The diagram shows Figure 3A A partial cross-sectional view of another example of a similar diagnostic cartridge, wherein an oxidant is incorporated into the membrane layer. According to embodiments provided herein, the membrane layer can be homogeneous or heterogeneous.

[0017] Figure 3C A partial cross-sectional view of one example of a modified oxygen sensor according to embodiments provided herein is illustrated, wherein an oxidant is incorporated into the membrane.

[0018] Figure 3D A schematic top view of one example of a diagnostic box including a primary oxygen sensor and a modified oxygen sensor as well as a reference sensor and other additional sensors and a common ground is illustrated according to embodiments provided herein.

[0019] Figure 3E A partial cross-sectional view of one example of a modified oxygen sensor according to embodiments provided herein is illustrated, wherein an oxidant is incorporated into the outermost layer of a multilayer film.

[0020] Figure 4 A schematic diagram of one example of a diagnostic analyzer including a diagnostic cartridge containing a sensor assembly receivable therein and illustrating its sensor components and their connection to a controller of the diagnostic analyzer according to embodiments provided herein is illustrated.

[0021] Figure 5 Illustrated is a bottom plan view of another example of a diagnostic cartridge according to embodiments provided herein, the cartridge including a sensor assembly configured to provide measurements for use in detecting hemolysis levels, wherein a sample pathway thereof includes a plurality of branches or offshoots that may branch from a main pathway.

[0022] Figure 6 The diagram shows an embodiment according to the present invention. Figure 5 6-6 of FIG. 1 , which is a cross-sectional view of a first passage portion that may be branched from a main passage of an exemplary diagnostic cartridge.

[0023] Figure 7 The diagram shows an embodiment according to the present invention. Figure 5 7-7 of FIG. 7 is a cross-sectional view of a second passage portion that can be branched from the main passage of an exemplary diagnostic box.

[0024] Figure 8 A flow chart illustrating an exemplary method of detecting hemolysis using a diagnostic sensor assembly and a diagnostic analyzer according to an embodiment of the present disclosure is illustrated. DETAILED DESCRIPTION

[0025] Regardless of how the grammatical term is used, the term includes individuals with male, female, or other identities.

[0026] In some embodiments, a diagnostic sensor assembly is provided. The diagnostic sensor assembly can be implemented in a diagnostic cassette (e.g., a card-shaped component) comprising a cassette body containing pathways and a plurality of sensors. For example, in some embodiments, the diagnostic cassette can be a disposable cassette. The diagnostic cassette can be received by a diagnostic analyzer (e.g., inserted into or otherwise coupled to the cassette), which is configured to provide blood analysis and, in particular, to detect (quantify) the level of hemolysis in a blood sample. For example, the diagnostic analyzer can be a handheld or desktop diagnostic analyzer. A blood sample can be provided to the diagnostic sensor assembly of the diagnostic cassette. For example, the blood sample can be whole blood. Alternatively, the sample can be serum or plasma, which may contain hemolysis. The amount of whole blood used for the test can be very small, e.g., 100 μL or less.

[0027] When the sensor assembly is included in a diagnostic cartridge, a cartridge body configured for connection to a diagnostic analyzer can be provided in any suitable configuration to provide measurement signals from the various sensors of the diagnostic cartridge. The cartridge body can include a diagnostic sensor assembly having a sample inlet configured to receive a sample therein, and a sample passageway extending from the sample inlet, for example, into the cartridge body. The sample (e.g., whole blood) can be provided (e.g., injected) from the sample inlet into the sample passageway, for example, using a syringe, a pumping mechanism, or other suitable sample transfer device.

[0028] The diagnostic sensor assembly of the diagnostic cartridge further includes a plurality of sensors. In the embodiment described herein, the sensor assembly includes two oxygen sensors. Specifically, the sensor assembly includes a primary oxygen sensor configured to contact a sample along a sample path, and a modified oxygen sensor also configured to contact the sample along the sample path. The modified oxygen sensor differs from the primary oxygen sensor in that the modified oxygen sensor includes (i.e., contains or is associated with) an oxidant, wherein the oxidant is configured to oxidize extracellular hemoglobin in the sample to methemoglobin (Fe 2+ to Fe 3+ ). In particular, in some embodiments, the modified oxygen sensor includes an oxidant such that it is provided in the membrane of the modified oxygen sensor. In other embodiments, the modified oxygen sensor includes an oxidant such that the oxidant is not located in the membrane, but rather is located in the sample pathway proximal to (e.g., upstream of) the oxygen sensor, thereby enabling the sample to flow through the oxidant before flowing through the oxygen sensor. In this manner, extracellular hemoglobin is oxidized before reaching the proximal oxygen sensor. Thus, this embodiment involves measuring a "modified" oxygen level in the sample, and the term "modified oxygen sensor" therefore also refers to the combination of the upstream oxidant and the proximal oxygen sensor. The amount of sample modification will depend on the extent and presence of hemolysis within the sample due to upstream reactions of the oxidant provided in the pathway with the sample.

[0029] In each embodiment, a quantitative determination of the hemolysis level of a sample can be obtained based on the signals generated by the primary and modified oxygen sensors. These signals indicate the corresponding local O2 measurements near the primary and modified sensors of the diagnostic sensor assembly. These signals can be provided to a controller of the diagnostic analyzer for processing and making a hemolysis level determination.

[0030] Due to the presence of the oxidant, if free (extracellular) hemoglobin is present in the sample, oxidation occurring near the modified oxygen sensor releases oxygen and, as a result, results in a higher level of local oxygen sensed by the modified oxygen sensor than by a primary oxygen sensor that does not include the oxidant. The oxidant is configured to contact the sample. The difference between the oxygen signals of the primary and modified oxygen sensors can be correlated to the level of hemoglobin present in the sample and, therefore, can provide a measure (quantification) of the level of hemolysis in the sample. The hemolysis level can be processed to provide a hemolysis index. For example, this index can range from zero to a maximum value and can be displayed to an operator of the diagnostic analyzer or otherwise communicated electronically within a hospital information system. Calibration can be performed before (or even after) running a hemolysis test to ensure that the diagnostic sensor assembly will produce correct results.

[0031] In another embodiment, a diagnostic analyzer is provided that is configured to provide for the detection (quantification) of hemolysis in a blood sample. The diagnostic analyzer comprises an analyzer body that includes a cartridge receiver and an electrical connector. The electrical connector is configured to establish an electrical connection between a controller and a diagnostic sensor assembly that is disposed within a diagnostic cartridge receivable within the cartridge receiver. The cartridge body is configured to be received by the cartridge receiver, wherein the cartridge body includes electrical contacts that are configured to couple to the electrical connector when the cartridge body is received by the cartridge receiver. The diagnostic sensor assembly of the cartridge body, including a sample inlet and a sample passage, is configured to receive a sample therein.

[0032] The primary oxygen sensor is configured to contact a sample provided in the sample path and is configured to provide a first measurement of the sample (a signal related to the amount of oxygen in the vicinity of the primary oxygen sensor). Similarly, the modified oxygen sensor is configured to contact a sample provided in the sample path, and the oxidant (of the modified oxygen sensor) is operable to oxidize extracellular hemoglobin to methemoglobin (from Fe 2+ to Fe 3+ ). Thus, the modified oxygen sensor is configured to provide a second measurement (a signal related to the amount of oxygen in the sample proximate the modified oxygen sensor).

[0033] The controller of the diagnostic analyzer is electrically coupled to an electrical connector (of the analyzer body). The electrical connector may include multiple conductive paths for connecting to multiple electrical contacts (of the sensor assembly) and transmitting sensor information from each sensor of the diagnostic sensor assembly. Specifically, the controller is configured to receive a first measurement result and a second measurement result via the connection between the electrical connector and the electrical contacts of the diagnostic sensor assembly.

[0034] The controller is further configured to provide a (quantified) hemolysis level measurement based on the first and second measurements. Specifically, this processing is performed by a processor of the controller, wherein its hemolysis detection module is configured to execute a difference finding routine. The difference between the first and second measurements, obtained by subtraction, is correlated with the degree of hemolysis present in the sample. Specifically, an elevated second measurement compared to the first measurement is quantitatively correlated with the degree of hemolysis (index level) of the sample, wherein a higher index level indicates a greater amount of hemolysis in the sample.

[0035] In yet another embodiment, a method for detecting hemolysis of a sample (e.g., whole blood, plasma, or serum) is provided. The method includes coupling a diagnostic sensor assembly to a diagnostic analyzer. The sensor assembly may be implemented as part of a diagnostic cartridge that is coupleable to, i.e., configured to be coupled to, a cartridge receiver of the diagnostic analyzer. The sensor assembly of the diagnostic cartridge may include: a sample inlet configured to receive a sample; a sample path extending from the sample inlet (e.g., into a cartridge body); a primary oxygen sensor configured to contact the sample in the sample path; and a modified oxygen sensor configured to contact the sample in the sample path, wherein the modified oxygen sensor includes a sensor configured to oxidize extracellular hemoglobin in the sample to methemoglobin (Fe 2+ to Fe 3+ ) oxidant. The method also includes passing the sample through a sample path and into contact with the primary oxygen sensor and the modified oxygen sensor, and obtaining a first measurement of the sample from the primary oxygen sensor and a second measurement of the sample from the modified oxygen sensor. These measurements may include signals related to local oxygen readings. The first measurement is related to a first partial pressure measurement of oxygen in the sample near the primary oxygen sensor. The second measurement is related to a second partial pressure measurement of oxygen in the sample near the modified oxygen sensor. According to the method, a hemolysis level is determined based on the first and second measurements, quantifying the extent of hemolysis contained in the sample (an index level).

[0036] This article refers to the Figure 1-8 These and other embodiments of the present disclosure are fully described below.

[0037] refer to Figure 1 , a perspective view of an example of a diagnostic analyzer 100 is shown according to an embodiment provided herein. The diagnostic analyzer 100 includes an analyzer body 102, which is configured to house various user interfaces, such as user control tactile devices (e.g., buttons, switches, touch screen, etc.). In the depicted embodiment, the analyzer body 102 of the diagnostic analyzer 100 may include a computing device 104. In some embodiments, the computing device 104 may be detachably mounted to a device mount 102M of a base 102B of the diagnostic analyzer 100. The computing device 104 may be a handheld computing device, such as a personal digital assistant (PDA), a tablet computer, or other similar computing device. In some diagnostic analyzers, the processing and storage functionality of the computing device 104 may be housed internal to the analyzer body 102, rather than as a detachable / removable version of the computing device 104.

[0038] The diagnostic analyzer 100 includes a controller 105. In this embodiment, the controller 105 is configured to include a first controller 105C1 and a second controller 105C2. The first controller 105C1 may be part of the base 102B of the analyzer body 102, and the second controller 105C2 may be part of or integrated with the computing device 104. The first controller 105C1 and the second controller 105C2 are in electronic communication with each other and can perform different functions. In this embodiment, the computing device 104 may include a display 104D to enable user input and visual display of operational information, test results, and other information. In some embodiments, the display 104D is tiltable about a pivot axis 102A. For example, a device mount 102M of the base 102B may receive the computing device 104 and pivot about the pivot axis 102A at a position 102L to allow adjustment of the viewing angle.

[0039] The display 104D may be a touch screen having a user interface that allows an operator to control the operation of the diagnostic analyzer 100, view measurement results generated by the diagnostic tests performed, and / or other auxiliary functions in conjunction with one or more tactile devices (e.g., buttons, switches, or other user-controlled devices). The first controller 105C1 may include electronics that enable communication with the diagnostic sensor assembly 103 implemented in the diagnostic cartridge 106, including signal conditioning (including filtering, A / D conversion, and / or possible amplification) of the various sensor signals received from the various sensors of the diagnostic sensor assembly 103 of the diagnostic cartridge 106, as will be described in greater detail herein. The first controller 105C1 may also include electronics that enable providing a current or potential input to the diagnostic sensor assembly 103 as a baseline input for carrying a signal related to the sensor measurement signals.

[0040] The second controller 105C2 is operable to perform the process and may include a hemolysis detection module 430 ( Figure 4 ), the hemolysis detection module 430 is a software module that includes a difference-finding routine. The software module that includes the difference-finding routine can be stored in the memory 431 of the diagnostic analyzer 100 and executed on the processor 432. The hemolysis detection module 430 receives sensor readings (signals) from the various oxygen sensors described herein and compares their values ​​to obtain the difference between them. Based on this difference, the hemolysis detection module 430 can effectively detect the level of hemolysis contained in the sample 111. Signals from any additional sensors in the diagnostic sensor assembly 103 can also be received and processed. In some embodiments, the test results and other information can be transmitted to the hospital information system (HIS) 101.

[0041] Reference again Figure 1, the diagnostic analyzer 100 may include a cartridge receiver 102P, which may be a port, opening, or other suitable coupling feature configured to receive and couple to a diagnostic cartridge 106. The diagnostic cartridge 106 may be coupled or connected to (e.g., inserted into) the cartridge receiver 102P, and by doing so, an electrical connection may be established between the diagnostic sensor assembly 103 and the controller 105 to allow processing. As shown, the cartridge receiver 102P may include a notch sized to receive the diagnostic cartridge 106 therein. The diagnostic cartridge 106 may be similar to a playing card in that it is thin compared to its width and length. For example, the diagnostic cartridge 106 may have a length of approximately 85 mm, a width of approximately 55 mm, and a thickness of approximately 1.2 mm. However, the diagnostic cartridge 106 may include other sizes and / or shapes.

[0042] like Figure 1 As shown in FIG, the diagnostic sensor assembly 103 can be configured as part of a cartridge body 108 of a diagnostic cartridge 106 configured to be connected to a diagnostic analyzer 100. The diagnostic sensor assembly 103 includes a sample inlet 109, which can be a port, an opening, a receiving element, etc., configured to receive a sample 111 to be tested therein. The sample inlet 109 can be disposed on a top layer 108T of the cartridge body 108, as shown in FIG. Figure 1 The sample inlet 109 may include a circular or other shaped opening, thereby providing a port configured to receive a sample 111 therein. In some embodiments, the sample inlet 109 may have a width or diameter dimension of about 4 mm to about 8 mm, although other diameters, sizes, or shapes may also be used.

[0043] The sample 111 may be whole blood, and the sample inlet 109 may be configured to allow a syringe or other suitable pump or transfer device to be sealingly coupled to the sample inlet 109 to receive the sample 111 therein and allow the sample 111 to be injected and flow into the sample passage 210 ( Figure 2 ). The cartridge body 108 may be made of multiple layers of material adhered together to form the sample passage 210 therein. For example, the cartridge body 108 may include a bottom layer 208B ( Figure 2 )、Top floor 108T( Figure 1 ) and possibly the intermediate layer 308I ( Figures 3A-3Cand 3E), which may be coupled together and sealed by any suitable means, such as by using an adhesive, mechanical coupling, combinations thereof, or the like. A portion of the sample passage 210 may be formed by the interaction of mating portions of the top layer 108T, the bottom layer 208B, and the middle layer 308I (if present). In some embodiments, the top layer 108T, the bottom layer 208B, and the middle layer 308I may be formed from the same type of material. For example, the material may be a plastic, such as polypropylene. Alternatively, different materials (e.g., different types of plastics, paper, foil, and / or laminates thereof) may be used for the middle layer 108I, the top layer 108T, and the bottom layer 208B. In some embodiments, the top layer 108T and / or the bottom layer 208B may be a transparent (transparent or translucent) material, allowing visual observation of the flow of the sample 111 therethrough. Other configurations for forming the sample passage 210 may be used, such as other two-piece, three-piece, or other multi-piece designs.

[0044] Reference again Figure 2 , shows a bottom view of the diagnostic cartridge 106 including the diagnostic sensor assembly 103. In the depicted embodiment, the sample passage 210 may include a first portion 210A extending from the sample inlet 109 to a second portion 210B. The second portion 210B may include a sensor array 212 comprised of a plurality of sensors, including at least the two oxygen sensors. In some embodiments, the sample passage 210 may extend from the sample inlet 109 into the cartridge body 108. The second portion 210B may have different dimensions than the first portion 210A. For example, the second portion 210B may be wider to accommodate the dimensions of the various sensors 214, 216, and 217 housed therein. Thus, in some embodiments, the second portion 210B may resemble a chamber. A waste passage 219 may be coupled to the downstream end of the second portion 210B. The waste passage 219 comprises a conduit or channel configured to receive sample outflow after the sample 111 contacts the last sensor or component (e.g., the additional sensor or ground 217) in the sensor array 212.

[0045] For example, the sample passage 210 may have a diameter from 12,500 µm 2 Up to 0.8 mm 2 In some embodiments, the sample passage 210 may have an aspect ratio W:H that may be approximately 5:1 or greater. The height H is the dimension across the sample passage 210, such as Figure 3A As shown in FIG, the width W spans the sample passage 210, as shown in FIG. Figure 2. The width W may be from about 250 μm to about 2 mm, and the height H may be from about 50 μm to about 400 μm. The length L along the sample path 210 from the sample inlet 109 to the start of the waste path 219 may be from about 1.25 mm to about 100 mm or more. Other relationships between the length L, width W, and / or height H may be employed, and other suitable length L, height H, and / or width W dimensions may be used.

[0046] In more detail, the sensor array 212 of the diagnostic sensor assembly 103 includes a primary oxygen sensor 214 configured to contact the sample 111 along the sample path 210 and a modified oxygen sensor 216, 316 also configured to contact the sample 111 along the sample path 210. Both of the oxygen sensors 214, 216, or 316 may be disposed in the second portion 210B, as shown. Figure 2-3C and Figure 3E As shown in the depicted embodiment of FIG. The modified oxygen sensor 216, 316 may be located downstream of the main oxygen sensor 214 (e.g., Figure 2 , on the left), such that the oxidant 324 ( Figures 3A-3C and 3E) will not alter the sample 111 exposed to the primary oxygen sensor 214. Other additional sensors and / or ground 217 may be provided between the primary oxygen sensor 214 and the modified oxygen sensors 216, 316, or otherwise located within the sensor array 212.

[0047] Modify the oxygen sensor 216, 316 to include an oxidizer 324 ( Figures 3A-3C and 3E), which are configured to oxidize extracellular (free) hemoglobin to methemoglobin. In particular, the oxidant 324 is configured to oxidize the iron of the extracellular hemoglobin from the ferrous state (Fe +2 ) is oxidized to the ferric state (Fe +3 ). In each of the depicted embodiments, Figures 3A-3C Partial cross-sectional views are shown in 3 and 3E.

[0048] exist Figure 3A In an embodiment, the oxidant 324 may be implemented as part of the membrane 316M. The membrane 316M may be formed from any semipermeable, wettable material, such as a polymeric material. For example, the polymeric material may be a polyurethane-based material, a polyacrylate-based material, copolymers thereof, and the like. The sensor chamber 316C formed at least in part by the membrane 316M of the modified oxygen sensor 216 may be provided with an electrolyte 316E disposed within the sensor chamber 316C.

[0049] Electrolyte 316E can be any solid proton-conducting polymer, such as a mixture of Nafion™ and polyvinylpyrrolidone, which can be mixed in a 4:1 ratio, for example. Alternatively, for example, electrolyte 316E can be a hydrogel comprising poly-N-vinylpyrrolidone K90 (PNPV) and 2,6-bis(4-azidobenzylidene)-4-methylcyclohexanone. Other suitable liquid or gel electrolytes suitable for this pump-type and Clark-type oxygen sensor can be used.

[0050] The oxidizing agent 324 may be incorporated into the polymer material forming the membrane 316M. The oxidizing agent 324 may be homogeneously incorporated into the membrane 316M, or alternatively, may be incorporated in a gradient state, i.e., with a concentration gradient, wherein a higher concentration of the oxidizing agent 324 may be provided at the surface of the membrane 316M adjacent to the sample 111 located in the second portion 210B of the sample passage 210. For example, the gradient may be driven by depositing multiple layers of the same polymer material with varying levels of the oxidizing agent 324 present, with the highest concentration being provided at the outermost layer adjacent to the sample 111.

[0051] In principle, film 316M comprising the desired film substrate and including oxidant 324 can be formed by any suitable process, such as deposition of a volatile liquid. In such deposition, the liquid mixture can be dispensed from a tip. Alternatively, film 316M can be formed by spin coating, dip coating, screen printing, spray coating, etc.

[0052] exist Figure 3B and 3C In the embodiment of FIG, the oxidant 324 may be incorporated into the matrix of the heterogeneous membrane 316M, as shown in the modified oxygen sensor 316. Figure 3B and 3C In this heterogeneous embodiment, membrane 316M is disposed in direct contact with sample 111 and is located between sample 111 and one or more electrodes 225. For illustrative purposes, the one or more electrodes 225 are shown as a single electrode (e.g., a working electrode). A reference electrode may be located at a different location. However, it should be understood that in some embodiments, the one or more electrodes 225 may consist of a working electrode and a counter electrode and / or a reference electrode. Any conventional electrode configuration or arrangement may be used.

[0053] Membrane 316M may include a hydrophobic polymer mixed with a hydrophilic component. For example, membrane 316M may include a heterogeneous membrane composition having a hydrophilic electrolyte-containing compartment and a hydrophobic compartment that supports the transport of gases (e.g., O2) and water vapor. The hydrophobic compartment may include a polymer.

[0054] Exemplary polymeric materials for the membrane 316M may include polysiloxanes, polyorganophosphazenes, poly-1 trimethylsilyl-1 propyne, poly-4-methyl-2-pentyne, and mixtures thereof. For example, the hydrophilic component of the mixture may include a hydrophilic polymer such as polyvinyl alcohol (PVA), a polyacrylate polymer (e.g., hydroxymethylacrylate), polyacrylamide, a polysaccharide, a cellulosic polymer, and / or gelatin. The hydrophilic component may also include some or all of the following: an emulsifier, a hydrophilic polymer binder, an electrolyte salt, a viscosity modifier, and other optional solubilized components. Other optional components of the hydrophilic compartment may include one or more components, such as a cross-linking agent, a catalyst, a redox agent, a buffer, and / or a surfactant, which may be incorporated into the membrane 316M during formation.

[0055] The gas diffusion coefficients of the phases should differ by 10 or more, 50 or more, or even 100 or more, with the hydrophobic component being significantly higher. In one approach, for example, the membrane 316M and the oxidant 324 can be deposited in a well formed in the bottom layer 208B. The bottom layer 208B can be an electrically insulating material, such as an epoxy resin layer, a polymer composite material, or other suitable electrically insulating material. The film-forming solution can be dried to form the membrane 316M comprising the heterogeneous membrane. Further discussion of the construction and materials of such conventional heterogeneous sensors can be found in US 7,094,330.

[0056] exist Figure 3E In an embodiment, sensor 316 may include a membrane 316M comprising multiple layers. For example, base layer 316B may be any oxygen-permeable, non-wettable material, such as polyethylene or polytetrafluoroethylene (PTFE), silicone, paraffin, etc. Top layer 316T is configured to contact sample 111 and may be made of a wettable material, such as a polyurethane-based material, a polyacrylate-based material, or copolymers thereof. The wettable material comprising top layer 316T contains an oxidizing agent 324 therein.

[0057] Figure 3A The membrane 316M of the embodiment and Figure 3EThe wettable material of the top layer 316T of an embodiment is hydrophilic and may include a contact angle greater than 0 degrees and less than or equal to 90 degrees. In some embodiments, the surface of the wettable material may have a contact angle less than 45 degrees or even less than 30 degrees. In some embodiments, the surface of the wettable material film 316M and the top layer 316T may be modified to further enhance their wettability and the availability of the oxidizing agent 324.

[0058] For example, the top surface of the membrane 316M may be treated in some manner ( Figure 3A ) and the top surface of the top layer 316T ( Figure 3E ) to enhance wettability. For example, the top surface can be plasma treated, i.e., treated with ionized gases and / or free radicals, for a sufficient time (e.g., 10 seconds to 5 minutes) to achieve a lower contact angle. Alternatively, the top surface can be treated with ultraviolet ozone (UVO) for an appropriate time (e.g., approximately 5 minutes) to achieve a lower contact angle. Other suitable treatments for enhancing wettability, as well as combinations of the aforementioned contact angle-reducing treatments, can be used.

[0059] Wettability can be measured by an optical tensiometer using the sessile drop method. In some embodiments, the oxidant 324 can be provided at a higher concentration in the outer portion of the top layer 316T adjacent to the sample 111 in order to maximize the amount of oxidant 324 available to oxidize any extracellular hemoglobin contained in the sample 111.

[0060] In all embodiments described herein, the oxidant 324 included in the modified oxygen sensor 316 may include potassium ferricyanide, for example. Alternatively, the oxidant 324 may include any chemical compound from other known classes of oxidants, such as organic nitrates or inorganic nitrites, aromatic amines, or quinones, which operate to cause oxidation of extracellular hemoglobin in the sample 111 to methemoglobin (Fe 2+ to Fe 3+ The oxidant 324 can be provided in an effective amount to cause the extracellular hemoglobin in the sample 111 to be fully oxidized to methemoglobin (Fe 2+ to Fe 3+ ). The goal is to have a sufficient concentration of oxidant 324 available to oxidize the hemoglobin to provide a sufficient difference in sensing readings between the primary oxygen sensor 214 and the modified oxygen sensors 216, 316. In particular, the oxidant 324 used should not interfere with the readings of any other sensor or sensors along the sample path 210, 310. In some embodiments, for example, Figures 3B-3CIn the embodiment shown in , one or more additional thin layers of membrane 316M may be formed adjacent to the one or more electrodes 225, which thin layers may have the same material as the membrane 316M, but without the oxidant 324, so that any deleterious redox type interactions at the one or more electrodes 225 may be minimized.

[0061] In any of the above embodiments, the oxidant 324 may be disposed within the membrane 316M, i.e., incorporated into the membrane 316M ( Figures 3A-3C and 3E). The oxidant 324 can be provided in a suitable amount, i.e., a suitable weight percentage (wt %), of about 0.01 wt % to about 25 wt %, or in some embodiments even from about 0.5 wt % to about 5.0 wt %, based on the total weight of the membrane 316M including the oxidant 324.

[0062] As will be appreciated, the oxidant 324 of the membrane 316M causes the oxidation of extracellular hemoglobin to methemoglobin (Fe 2+ to Fe 3+ ), which subsequently releases bound oxygen in a localized manner from extracellular hemoglobin in the sample 111. Thus, the released / generated oxygen can be detected by the modified oxygen sensor 216, 316, and thus, quantification of hemolysis of the sample 111 can be obtained in a differential manner (e.g., as the difference between the readings from the primary oxygen sensor 214 and the modified oxygen sensor 216, 316) when free (extracellular) hemoglobin is present in the sample 111, as will be further explained herein.

[0063] The primary oxygen sensor 214 is configured to provide a first measurement related to the partial pressure of oxygen in the sample 111 near the primary oxygen sensor 214. Similarly, the modified oxygen sensor 216, 316 is configured to provide a second measurement related to the partial pressure of oxygen in the sample 111 near the modified oxygen sensor 216, 316. In a sample 111 having free hemoglobin therein, the oxidation reaction between the extracellular hemoglobin in the sample 111 and the oxidant 324 will release excess oxygen, which can then be sensed locally by the modified oxygen sensor 216, 316. A difference measurement can then be obtained compared to the reading from the primary oxygen sensor 214.

[0064] Reference again Figure 3C and 3E , shows an example of a modified oxygen sensor 316 having a solid-state integrated chip structure. The modified oxygen sensor 316 can be provided in the diagnostic box 306, such as Figure 3D , which in use may be connected to a diagnostic analyzer 100 ( Figure 1 )'s inlet 351 and outlet 352, or is otherwise connected to the sample passage 210 and the waste passage 219. Figure 3C and 3E A modified oxygen sensor 316 may be included as Figure 2 in the diagnostic box shown in .

[0065] like Figure 3D As shown in FIG, inlet 351 supplies sample 111 to sensor array 212 including primary oxygen sensor 214 and modification sensor (MO2) 316. Diagnostic cartridge 306 may also include one or more additional sensors and / or ground 217. Additional sensors may be configured to measure other analytes and / or conditions, such as Cl-, Mg++, Na+, K+, pCO2, Ca++, glucose, lactate, creatinine, etc. Additional sensors 217 configured to sense other analytes (e.g., BUN, Hct, and / or TCO2) and / or conditions (e.g., pH) may also be included or alternatively included.

[0066] The diagnostic box 306 may also include one or more reference sensors 354 configured to provide a reference signal. Alternatively, the reference signal may be obtained within the diagnostic analyzer 100 or at other locations along the sample path 210 or 310. The diagnostic box 306 may also include a common ground at any suitable location that can be connected to the controller 105. The arrangement of the sensors may differ from that shown. However, the modified oxygen sensor 316 may be located downstream of the main oxygen sensor 214. In addition, the modified oxygen sensor 316 and the main oxygen sensor 214 may be separated by one or more additional sensors 217 or a suitable space so that the main oxygen sensor 214 is not affected by the additional oxygen released near the modified oxygen sensor 316.

[0067] The sensors 214, 316, 217 and the one or more reference sensors 354 and ground (if used) may be electrically coupled to the diagnostic analyzer 100 ( Figure 1 and Figure 4 ) detection system, which may include any suitable electronics to enable reading of the potential difference (or current difference) between the primary oxygen sensor 214 and the modified oxygen sensor 216, 316 as a measurable signal. Again, the construction of the detection system and reference sensor 354 is well known and will not be further described herein. For example, the reference system and reference sensor 354 may be of the type used in the epoc® blood analysis system available from Siemens Medical Solutions, or as Figure 3D However, the reference sensor 354 may be positioned at other locations along the sample path 210 , 310 besides on the second portion 210B, 310B.

[0068] In more detail and with further reference Figure 3C and 3E, the modified oxygen sensor 316 may include a box body 308 including an oxygen permeable membrane 316M coupled thereto (e.g., coupled to the bottom layer 208B). The wall 356 of the diagnostic box 306 and the sensors (e.g., the primary oxygen sensor 214, the modified oxygen sensor 316, the reference sensor 354 (see Figure 3D 、 3E 4) and any additional sensors 217) are formed in a second portion 310B in which the sample 111 is received. Membrane 316M may be formed as a thin polymer sheet that is selective for O2. For example, membrane 316M may have a diameter or maximum dimension ranging from approximately 200 μm to approximately 1,700 μm, and a thickness ranging from approximately 10 μm to 200 μm. Other suitable diameters or maximum dimensions and / or thicknesses may be used. The chemical composition of membrane 316M that renders it selective for O2 may be as described above herein, or may be any other suitable oxygen-permeable material.

[0069] Modifying the oxygen sensor 316, the reference sensor 354, and any additional sensors and / or ground 317 may include one or more electrodes 225, which in some embodiments may be positioned adjacent to the membrane 316M. The one or more electrodes 225, which may include a working electrode, a counter electrode, and / or a reference electrode, may be made of any suitable construction, such as conductive traces, mask deposition, etc. A conductor may extend from each of the one or more electrodes 225, the reference sensor 354, and the other additional sensors and / or ground 317 to a corresponding electrical contact (e.g., Figure 2 and 4 218 ), the electrical contacts may be disposed on the cartridge body 108 , 308 of the diagnostic cartridge 106 , 306 (e.g., on the bottom thereof), wherein the electrical contacts 218 may be interconnected with the diagnostic analyzer 100 when the diagnostic cartridge 106 , 306 is coupled to the diagnostic analyzer 100.

[0070] For example, the one or more electrodes 225 may comprise silver (AG) elements, which may be coated with, for example, a silver chloride (AGCl) coating. Alternatively, for example, the one or more electrodes 225 may comprise gold or platinum, or any combination thereof. Other suitable conductive materials or combinations of conductive materials may be used. The connection between the one or more electrodes 225 and the electrical contacts 218 provided on the box body 108, 308 of the diagnostic box 306 may be made of any suitable conductive material and may be a trace or conductor formed in any suitable configuration, such as by printing, deposition, or other known conductive conduit or trace formation methods.

[0071] In some embodiments, the sensor array 212, 312 may include one or more additional sensors 217 in addition to the main oxygen sensor 214 and the modified oxygen sensors 216, 316. For example, the other sensors 217 may be configured to sense other analytes and / or conditions as described above. In some embodiments, the number of additional sensors 217 may be one or more, five or more, or even ten or more. The total number of sensors in the sensor array 212, 312 may range from 2-15, for example, including the main oxygen sensor 214 and the modified oxygen sensors 216, 316. Figure 3A 、 3B As shown in and 3D , one or more additional sensors 217 may be positioned in the second portion 210B, 310B between the primary oxygen sensor 214 and the modified oxygen sensor 216 , 316 .

[0072] In some embodiments, as Figure 2 , the cartridge body 108 of the diagnostic cartridge 106 may include a control portion 220 that may be used to supply a calibrator liquid to the sample passage 210 in order to calibrate the operation of the diagnostic sensor assembly 103 of the diagnostic cartridge 106. Such a control portion 220 is known to those skilled in the art and will not be further described herein.

[0073] Reference again Figure 1-4 Operation of the diagnostic analyzer 100 in the depicted embodiment may involve inserting the diagnostic cartridge 106, 306 into the cartridge receiver 102P to engage the diagnostic sensor assembly 103, 303 of the diagnostic cartridge 106, 306 with the controller 105 via electrical connections between the diagnostic sensor assembly 103, 303 and the controller 105. The sample inlet 109 receives a sample 111 (e.g., contained in a syringe or other delivery vehicle), and the sample 111 flows into the sample passage 210, 310 to deliver the sample 111 to the second portion 210B, 310B containing the primary oxygen sensor 214 and the modified oxygen sensor 216, 316, as well as to any additional sensors 217 that may be included in the sample passage 210, 310. When the sample 111 contacts the oxygen sensor 214, 216, 316, a signal related to the oxygen level sensed by the oxygen sensor 214, 216, 316 is provided to the controller 105. The controller 105 may then process the various received sensor signals to generate a sensed level of the sensed analyte, state, or component (e.g., O2) for each sensor 214, 216, 316, and 217. Based on the sensed signals from the primary oxygen sensor 214 and the modified oxygen sensors 216, 316, the controller 105 may be operable to detect the level of hemolysis in the sample 111.

[0074] After passing through the second portion 210B, 310B, waste sample fluid can flow into the waste passage 219 for storage. For example, the waste passage 219 can be formed as a groove, recess, or similar structure within the cartridge body 108 of the diagnostic cartridge 106, 306, and in some embodiments, can have a serpentine or other non-straight shape. The waste passage 219 should be long enough to accommodate waste fluid (e.g., the sample 111 after passing through the second portion 210B, 310B and possible control fluid from the calibration operation of the control portion 220).

[0075] Reference again Figures 3A-3C , the diagnostic box 106, 306 may include one or more electrodes 225 for each sensor 214, 216, 217, 316 and 354, and each electrode is electrically connected to the electrical contact 218. Figure 2 For simplicity, only one electrical contact 218 is shown, but it should be understood that each sensor 214, 216, 217, 316, 354 has one or more dedicated electrical contacts for providing a signal, such as a voltage or current. Any potential or current changes resulting from the operation of the sensor can then be detected. If the sensor type is designed as a potentiometric type, a baseline voltage can be provided, and changes in this baseline voltage can be detected. For example, a low voltage from 1 mV to 500 mV or even from 1 mV to 50 mV can be provided as a baseline voltage in the measurement circuit. Similarly, if the sensor measurement system is an amperometric type, a baseline current can be provided, based on which changes can be detected. For example, a low current from 1 nA to 500 nA or even 1 nA to 50 nA can be provided as a baseline current in the measurement circuit. Other suitable voltage or current baseline values ​​can also be used. Any conventional circuit that enables measurement of voltage and / or current can be used.

[0076] like Figure 4 As shown in , for example, each of the one or more electrodes of the primary oxygen sensor 214, the modified oxygen sensor 216, 316, the additional sensor or ground 217, and the reference sensor 354 may have a conductive path extending to the controller 105. For example, in some embodiments, there may be a conductive path from and to a corresponding electrical contact 218 configured on a surface of the cartridge body 108, 308 of the diagnostic cartridge 106, 306. For example, as shown in Figure 4As shown in FIG, the conductive path may include connections to each of the electrical contacts 218A, 218B (only a few are labeled). The electrical contacts 218 are, in turn, contacted by the mating contacts 221 (e.g., mating contacts 221A, 221B, only a few are labeled) of the electrical connector 458 for connection to the controller 105. Similarly, the ground 217 (if used) may be connected to the contact 218G, which is contacted by the mating contact 221G of the electrical connector 458. These signals from the primary oxygen sensor 214 and the modified oxygen sensors 216, 316, as well as any optional additional sensors 217 and the reference sensor 354, can be effectively received by the controller 105. The electrical connection between the electrical contacts 218 and the mating contacts 221 can be established when the diagnostic cartridge 106, 306 is coupled to the cartridge receiver 102P.

[0077] Reference again Figure 4 The diagnostic analyzer 100 may also include a hemolysis detection module 430 within the controller 105. The hemolysis detection module 430 may be stored as programming code within the memory 431 and executed by the processor 432. The processor 432 may control the operation of the sensors 214, 216, 316, 354, 217, the memory 431, and / or the display 104D. The signals from each sensor 214, 216, 217 may be manipulated to provide a value related to the sensed analyte. In the case of the primary oxygen sensor 214 and the modified oxygen sensors 216, 316, the signals therefrom are manipulated to obtain a value related to their oxygen levels. Based on these signal values, the hemolysis detection module 430 may determine the level of hemolysis in the sample 111 based on the difference between them. This difference may be calculated using a difference finding program code. A larger difference found by the difference finding routine of the hemolysis detection module 430 indicates that more oxygen is being released due to the oxidation reaction with the oxidant 324 occurring in the vicinity of the modified oxygen sensor 216, 316, and the difference is correlated with the level of hemolysis (extracellular hemoglobin) present in the sample 111. The larger the difference determined or calculated by the difference finding routine, the greater the level or amount of hemolysis present in the sample 111.

[0078] Processor 432 may be any suitable computing resource, such as, but not limited to, a microprocessor, a microcontroller, an embedded microcontroller, a digital signal processor (DSP), a field programmable gate array (FPGA) configured to perform as a microcontroller, and the like.

[0079] Memory 431 may be any suitable type of memory, such as, but not limited to, one or more of volatile memory, non-volatile memory, or a combination thereof. Volatile memory may include, but is not limited to, static random access memory (SRAM) or dynamic random access memory (DRAM). Non-volatile memory may include, but is not limited to, electrically programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), flash memory, and the like. Memory 431 may store a plurality of instructions that, when executed by processor 432, cause processor 432 to perform various actions specified by one or more of the stored plurality of instructions (including the difference finding routine).

[0080] The user interface may include one or more display screens (e.g., display 104D). The user interface may be controlled by processor 432, and the functionality of the user interface may be implemented at least in part by computer-executable instructions (e.g., program code or software) stored in memory 431 and / or executed by processor 432 of diagnostic analyzer 100. In some embodiments, processor 432 may receive measurements from oxygen sensors 214, 216, 316, and (if one or more additional sensors, such as reference sensor 354, are included) additional sensor 217, process the measurements to generate calculations, and present the calculations and / or other information, such as patient information, via display 104D of the user interface. For example, the user interface and display 104D may be configured to present one or more measurements and / or calculations of hemolysis, and possibly other analyte and / or condition measurements, to a user of diagnostic analyzer 100. These results may also be transmitted to HIS 101.

[0081] Figure 5 An alternative embodiment of a diagnostic sensor assembly 503 that may be included in a diagnostic cartridge 506 is illustrated. This embodiment includes a sample pathway 510 similar to the previous embodiment, but the sample pathway 510 includes at least two bifurcations or branches, such as a first pathway 510B and a second pathway 510C. In some embodiments, the first pathway 510B and the second pathway 510C can optionally branch off from a main pathway 510A extending from the sample inlet 109. In this embodiment, the main oxygen sensor 214 can be located on the first pathway 510B and the modified oxygen sensor 516 can be provided on the second pathway 510C. For example, the main oxygen sensor 214 can be provided in the first pathway 510B branching off from the main pathway 510A and the modified oxygen sensor 516 can be provided in the second pathway 510C branching off from the main pathway 510A. For example, one or more optional additional sensors or grounds 217 of the same type described above can be provided on one or both of the first pathway 510B and the second pathway 510C. Other sensor types may be included alternatively or additionally. Likewise, in the embodiments described herein, Figure 5 In each embodiment, the reference sensor may be included anywhere along one or both of the first and second pathways 510B, 510C, or as part of the oxygen sensor 214, 214B. Again, for ease of illustration, the electrical contact 518 is shown as a single circle, but each of the main oxygen sensor 214, the oxygen sensor 214B, the additional sensor and / or ground 217, and the reference sensor 354 may have its own electrical contact. The diagnostic box 506 including the diagnostic sensor assembly 503 may be configured as described above. Figure 4 1 and connected to the diagnostic analyzer 100 in the same manner as shown in FIG.

[0082] Because the sample passage 510 is bifurcated and the two oxygen sensors 214, 516 are not located in series within the same single sample passage, there is advantageously reduced or no risk that the oxidant 524 associated with modifying the oxygen sensor 516 will alter the sample exposed to the primary oxygen sensor 214. Similar to before, the diagnostic cartridge 506 may include a control portion 220 and a waste passage 219 extending from each of the first passage 510B and the second passage 510C.

[0083] The difference of this embodiment of the modified oxygen sensor 516 is that the oxidant 524 of the modified oxygen sensor 516 is provided at a position in the second passage 510C so that the sample 111 can flow through the oxidant 524 and convert the extracellular hemoglobin from a ferrous state (Fe +2 ) is oxidized to the ferric state (Fe +3 ), which subsequently releases oxygen that can be sensed by oxygen sensor 214B. Oxygen sensor 214B may be identical to main oxygen sensor 214 located in first passage 510B and may have conventional construction. As shown, modified oxygen sensor 516 is comprised of or includes an oxidizer 524 and oxygen sensor 214B, where oxygen sensor 214B may be conventional, like main oxygen sensor 214. Oxidizer 524 may be positioned and located upstream of oxygen sensor 214B.

[0084] In an alternative embodiment, the modified oxygen sensor 516 may differ from the primary oxygen sensor 214 in the same manner as described above with respect to the modified oxygen sensors 216, 316, with the oxidant 324 contained within its membrane 316M. Figure 2 The embodiment of the present invention differs in that the modified oxygen sensor 516 can be located in a different branch of the sample path 510 (e.g., the second path 510C), and thus the first measurement result and the second measurement result can be obtained in any order. Figure 5In this alternative embodiment, the first and second measurements may be obtained simultaneously (or in any other order) depending on the position of the oxygen sensors in their respective pathways. The use of a bifurcated pathway can be used to reduce or minimize the risk of the modified oxygen sensor 516 altering the sample exposed to the primary oxygen sensor 214 and eliminate the requirement that the sample must flow through the primary sensor before flowing through the modified sensor.

[0085] In another embodiment, the oxidant 524 may be provided upstream of the oxygen sensor 214B in the second passage 510C and may also be provided in the membrane of the oxygen sensor 214B. The amount at each location may be adjusted to achieve the desired conversion of hemoglobin to methemoglobin (Fe 2+ to Fe 3+ ) for maximum oxidation.

[0086] Figure 6 and Figure 7 The diagram shows the Figure 5 6 and 7-7. The diagnostic box 506 can be constructed of multiple layers adhered together as before. Each of the first passage 510B and the second passage 510C can include an oxygen sensor 214, 214B, both of which can be conventional oxygen sensors and are the same as the main oxygen sensor 214 described above. The second passage 510C can include an oxidant 524. The oxidant 524 can be located at a position upstream of the oxygen sensor 514B. The oxidant 524 can be provided in any suitable form, such as in the form of a split cylinder or other shaped member, which can be sprayed on the passage portion of one or both of the top layer 108T and the bottom layer 208B or even on the middle layer 308I thereof and dried.

[0087] In this embodiment, the oxidant 524 may be provided with or without a binder material, such as a soluble water-based polymer material, such as polyvinyl alcohol, polyacrylic acid, or the like. The oxidant 524 may be mixed with the soluble water-based polymer material. The soluble water-based polymer material may be dissolved upon contact with the sample 111. The binder material may optionally be an insoluble material (e.g., cross-linked), but may be sufficiently wettable or porous to allow sufficient interaction with the sample 111, thereby facilitating oxidation of any extracellular hemoglobin present. In the case of an insoluble binder, this may allow for a reusable configuration, rather than a single-use application. In this configuration, the oxidant 524 may also be potassium ferricyanide. However, other suitable oxidant materials capable of sufficiently oxidizing hemoglobin may also be used.

[0088] The oxidizing agent 524 may be provided at approximately 0.01-99 wt % based on the combined weight of the binder and oxidizing agent 524. It may be desirable to have a very low amount of binder relative to the oxidizing agent 524. For example, some binders may be used at approximately 1 wt % to approximately 3 wt % as a means of simply "holding" the oxidizing agent 524 in place until dissolved by the sample 111. The thickness and length of the cylindrical or other shaped member including the oxidizing agent 524 may be of sufficient length and thickness or size to achieve a suitable measurable change in the oxygen level near the oxygen sensor 214B compared to the oxygen level at the primary oxygen sensor 214 when free extracellular hemoglobin is present.

[0089] In some embodiments, the oxidant 524 may be disposed in a well in the bottom layer 208B upstream of the oxygen sensor 214B, such as Figure 3B In the modified oxygen sensor 316, a well including the membrane 316M is provided. However, in this embodiment, the well need not include the electrode 225. The oxidant 524 may be incorporated into the membrane formed in the well or provided in some other form, for example, a form that can be dissolved by the sample as described above.

[0090] Figure 8 A flow chart is illustrated of one example of a method 800 for determining hemolysis of a sample 111 using a diagnostic sensor assembly 103, 303, 503 and a diagnostic analyzer 100 according to an embodiment of the present disclosure. The method 800 begins at block 802 by coupling a diagnostic cartridge 106, 306, 506 to a cartridge receiver 102P of the diagnostic analyzer 100. For example, the diagnostic cartridge 106, 306, 506 may be coupled to (e.g., inserted into) the cartridge receiver 102P of the analyzer body 102, which establishes an electrical connection between the diagnostic sensor assembly 103, 303, 503 of the diagnostic cartridge 106, 306, 506 and the controller 105.

[0091] In the depicted embodiment, the diagnostic sensor assembly 103, 303, 503 of the diagnostic cartridge 106, 306, 506 includes a sample inlet 109 configured to receive a sample 111; a sample passage 210, 310, 510 extending from the sample inlet 109 into the cartridge body 108, 308, 508; a primary oxygen sensor 214 configured to contact the sample 111 in the sample passage 210, 310, 510; and a modified oxygen sensor 216, 316, 516 configured to contact the sample 111 in the sample passage 210, 310, 510, the modified oxygen sensor 216, 316, 516 including an oxidant 224, 324, 524 configured to oxidize iron of extracellular hemoglobin to methemoglobin, as described above. In some embodiments, before using the diagnostic analyzer 100, the display 104D and / or user interface may prompt the user to couple the diagnostic cartridge 106, 306, 506, enter a user identification, scan a name tag or other barcode, enter a password or otherwise verify their identity, and / or provide a sample 111 into the sample inlet 109.

[0092] Once the cartridge body 108, 308, 508 has been coupled to the cartridge receiver 102P of the diagnostic analyzer 100, at block 804, the method 800 may further include passing the sample 111 through the sample passage 210, 310, 510 and into contact with the primary oxygen sensor 214 and the modified oxygen sensor 216, 316, 516. The sample 111 may also contact any additional sensors or ground 217 positioned along the sample passage 210, 310, 510, as well as the reference sensor 534. For example, a syringe or other sample delivery device may be employed to interface with the sample inlet 109 of the cartridge body 108, 308, 508. The syringe or other device may move the sample 111 into the sample inlet 109, through the sample passage 210, 310, 510, and into the waste passage 219.

[0093] Method 800 also includes obtaining a first measurement of sample 111 from primary oxygen sensor 214 at block 806 and obtaining a second measurement of sample 111 from modified oxygen sensor 216, 316, or 516 at block 808. At block 810, method 800 includes providing a hemolysis level in the sample based on the first and second measurements. This information may be collected by controller 105, and the hemolysis level may be displayed on a user interface (e.g., display 104D) to communicate to a user of diagnostic analyzer 100. Additionally, any other test results and other information, along with the hemolysis level, may be transmitted to hospital information system (HIS) 101 via a hardwired connection or wirelessly. For example, Wi-Fi or hardwired LAN communication may be used.

[0094] The sample 111 used for testing may be up to 100 μL or less, or in some embodiments even less than 50 μL or less, although other sample 111 amounts may also be used. As should now be apparent, the diagnostic sensor assembly 103, 303, 503 implemented in the diagnostic cartridge 106, 306, 506 may be used in the diagnostic analyzer 100 to perform multiple oxygen measurements to detect the level of hemolysis in the sample 111. As should also be appreciated, the diagnostic sensor assembly 103, 303, 503 may be included in the diagnostic analyzer rather than being implemented in the diagnostic cartridge and may include a cleaning system interfaced therewith to facilitate reuse of the diagnostic sensor assembly 103, 303, 503.

[0095] In some embodiments, the sensor array 212, 312 may be included solely as a disposable cartridge that can be coupled to a diagnostic analyzer, and the diagnostic analyzer itself may include a sample inlet and passage, a waste passage, controls, and a cleaning system that enables cleaning of the sensor array 212, 312 after use. Thus, the diagnostic cartridge may include an inlet (e.g., inlet 351) and an outlet (e.g., outlet 352) that sealingly connect to the inlet and waste passages of the diagnostic analyzer when the cartridge is coupled to the analyzer. Thus, in this case, the diagnostic cartridge includes the primary and modified oxygen sensors and electrical contacts as described herein, allowing the cartridge to be removed / detached from the diagnostic analyzer after multiple uses.

[0096] The following is a list of non-limiting illustrative embodiments disclosed herein: 1. A diagnostic sensor assembly comprising: a sample inlet configured to receive a sample; a sample passage extending from the sample inlet; a primary oxygen sensor configured to contact the sample along the sample path; and A modified oxygen sensor is configured to contact the sample along the sample pathway, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin, thereby enabling quantification of hemolysis of the sample.

[0097] 2. The diagnostic sensor assembly of illustrative embodiment 1, wherein the diagnostic sensor assembly is configured as part of a cartridge body of a diagnostic cartridge configured for connection to a diagnostic analyzer.

[0098] 3. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the primary oxygen sensor is configured to provide a first measurement related to a partial pressure of oxygen in the sample.

[0099] 4. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the oxidant of the modified oxygen sensor comprises potassium ferricyanide.

[0100] 5. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the oxidizing agent is configured to convert the hemoglobin from a ferrous state (Fe +2 ) is oxidized to the ferric state (Fe +3 ).

[0101] 6. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the modified oxygen sensor is configured to provide a second measurement related to the partial pressure of oxygen in the sample.

[0102] 7. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the oxidant is disposed in a membrane of the modified oxygen sensor.

[0103] 8. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the oxidant of the modified oxygen sensor is disposed at a location in the sample passageway such that the sample can flow through the oxidant and convert extracellular hemoglobin from a ferrous state (Fe +2 ) is oxidized to the ferric state (Fe +3 ).

[0104] 9. The diagnostic sensor assembly of any one of the preceding illustrative embodiments, wherein the main oxygen sensor is disposed in a first passage branching from a main passage, and the modification oxygen sensor is disposed in a second passage branching from the main passage.

[0105] 10. A diagnostic analyzer comprising: an analyzer body including a cartridge receiver and an electrical connector; a diagnostic cartridge capable of being received in the cartridge receiver, the diagnostic cartridge comprising: a box body configured to be coupled to the box receiver, the box body comprising: electrical contacts that are coupleable to the electrical connector when the box body is received in the box receiver, a sample inlet configured to receive a sample; a sample passage extending from the sample inlet into the cartridge body and configured to receive the sample therein; a primary oxygen sensor configured to contact the sample in the sample passage and configured to provide a first measurement of the sample; a modified oxygen sensor configured to contact the sample in the sample passage, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin and configured to provide a second measurement of the sample; and A controller is coupled to the electrical connector, the controller configured to receive the first and second measurements of the sample and to provide a level of hemolysis in the sample based on the first and second measurements.

[0106] 11. The diagnostic analyzer of illustrative embodiment 10, wherein the first measurement is related to a first partial pressure measurement of oxygen in the sample.

[0107] 12. The diagnostic analyzer of any one of the preceding illustrative embodiments, wherein the second measurement is related to a second partial pressure measurement of oxygen in the sample.

[0108] 13. The diagnostic analyzer of any one of the preceding illustrative embodiments, wherein a second measurement that is elevated compared to the first measurement is quantitatively correlated to a level of hemolysis in the sample.

[0109] 14. The diagnostic analyzer of any one of the preceding illustrative embodiments, wherein the controller is configured to determine a difference between the first measurement and the second measurement.

[0110] 15. The diagnostic analyzer of any one of the preceding illustrative embodiments, wherein the difference is quantitatively correlated to the level of hemolysis in the sample.

[0111] 16. A method for determining hemolysis of a sample, comprising: coupling a diagnostic cartridge to a cartridge receptacle of a diagnostic analyzer, the diagnostic cartridge comprising a sample inlet configured to receive the sample, a sample passage extending from the sample inlet into a cartridge body, a primary oxygen sensor configured to contact the sample in the sample passage, and a modified oxygen sensor configured to contact the sample in the sample passage, the modified oxygen sensor comprising an oxidant configured to oxidize extracellular hemoglobin to methemoglobin; passing the sample through the sample passage and into contact with the primary oxygen sensor and the modified oxygen sensor; obtaining a first measurement of the sample from the primary oxygen sensor; obtaining a second measurement of the sample from the modified oxygen sensor; and A level of hemolysis in the sample is provided based on the first measurement and the second measurement.

[0112] The foregoing description discloses only exemplary embodiments of the present disclosure. Modifications of the above-disclosed diagnostic cartridges, diagnostic analyzers, and methods for detecting hemolysis that fall within the scope of the present invention will be readily apparent to those skilled in the art. Therefore, it should be understood that other embodiments may also fall within the scope of the present invention, as defined by the following claims and their equivalents.

Claims

1. A diagnostic sensor assembly comprising: a sample inlet configured to receive a sample; a sample passage extending from the sample inlet; a primary oxygen sensor configured to contact the sample along the sample path; as well as A modified oxygen sensor is configured to contact the sample along the sample pathway, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin, thereby enabling quantification of hemolysis of the sample.

2. The diagnostic sensor assembly of claim 1, wherein: The diagnostic sensor assembly is configured as part of a cartridge body of a diagnostic cartridge configured for connection to a diagnostic analyzer.

3. The diagnostic sensor assembly of claim 1 , wherein: The primary oxygen sensor is configured to provide a first measurement related to a partial pressure of oxygen in the sample.

4. The diagnostic sensor assembly of claim 1 , wherein: The oxidant of the modified oxygen sensor includes potassium ferrocyanide.

5. The diagnostic sensor assembly of claim 1 , wherein: The oxidant is configured to convert the hemoglobin from a ferrous state (Fe +2 ) is oxidized to the ferric state (Fe +3 ).

6. The diagnostic sensor assembly of claim 1 , wherein: The modified oxygen sensor is configured to provide a second measurement related to a partial pressure of oxygen in the sample.

7. The diagnostic sensor assembly of claim 1 , wherein: The oxidant is disposed in a membrane of the modified oxygen sensor.

8. The diagnostic sensor assembly of claim 1 , wherein: The oxidant of the modified oxygen sensor is disposed at a position in the sample passageway such that the sample can flow through the oxidant and convert extracellular hemoglobin from a ferrous state (Fe +2 ) is oxidized to the ferric state (Fe +3 ).

9. The diagnostic sensor assembly of claim 8, wherein: The main oxygen sensor is provided in a first passage branching from a main passage, and the modified oxygen sensor is provided in a second passage branching from the main passage.

10. A diagnostic analyzer comprising: an analyzer body including a cartridge receiver and an electrical connector; a diagnostic cartridge capable of being received in the cartridge receiver, the diagnostic cartridge comprising: a box body configured to be coupled to the box receiver, the box body comprising: electrical contacts that are coupleable to the electrical connector when the box body is received in the box receiver, a sample inlet configured to receive a sample; a sample passage extending from the sample inlet into the cartridge body and configured to receive the sample therein; a primary oxygen sensor configured to contact the sample in the sample passage and configured to provide a first measurement of the sample; a modified oxygen sensor configured to contact the sample in the sample passage, the modified oxygen sensor including an oxidant configured to oxidize hemoglobin to methemoglobin and configured to provide a second measurement of the sample; and A controller is coupled to the electrical connector, the controller configured to receive the first and second measurements of the sample and to provide a level of hemolysis in the sample based on the first and second measurements.

11. The diagnostic analyzer of claim 10, wherein: The first measurement is related to a first partial pressure measurement of oxygen in the sample.

12. The diagnostic analyzer of claim 10, wherein: The second measurement is related to a second partial pressure measurement of oxygen in the sample.

13. The diagnostic analyzer of claim 12, wherein: A second measurement that is elevated compared to the first measurement is quantitatively correlated to the level of hemolysis in the sample.

14. The diagnostic analyzer of claim 10, wherein: The controller is configured to determine a difference between the first measurement and the second measurement.

15. The diagnostic analyzer of claim 14, wherein: The difference is quantitatively correlated to the level of hemolysis in the sample.

16. A method for determining hemolysis of a sample, comprising: coupling a diagnostic cartridge to a cartridge receptacle of a diagnostic analyzer, the diagnostic cartridge comprising a sample inlet configured to receive the sample, a sample passage extending from the sample inlet into a cartridge body, a primary oxygen sensor configured to contact the sample in the sample passage, and a modified oxygen sensor configured to contact the sample in the sample passage, the modified oxygen sensor comprising an oxidant configured to oxidize extracellular hemoglobin to methemoglobin; passing the sample through the sample passage and into contact with the primary oxygen sensor and the modified oxygen sensor; obtaining a first measurement of the sample from the primary oxygen sensor; obtaining a second measurement of the sample from the modified oxygen sensor; as well as A level of hemolysis in the sample is provided based on the first measurement and the second measurement.

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