Sample analyzer for analyzing urine sample, system, and method

Abstract

The present disclosure provides a sample analyzer for analyzing a urine sample, a system, and a method. The sample analyzer comprises: a sample preparation device; an optical detection device, comprising an optical detector; and a controller, configured to: control the sample preparation device to prepare a first test solution from a urine sample to be tested and a colorimetric reagent, and control the optical detector to test the first test solution so as to obtain a first urine total protein test result; control the sample preparation device to prepare a second test solution from said urine sample and a turbidimetric reagent, and control the optical detector to test the second test solution so as to obtain a second urine total protein test result; when said urine sample is proteinuria, on the basis of the first urine total protein test result and the second urine total protein test result, calculate a kidney injury marking parameter used for characterizing the kidney injury state of a subject; and output the kidney injury marking parameter.

Description

Sample analyzers, systems, and methods for analyzing urine samples

[0001] Cross-reference to Related Applications

[0002] The present disclosure is based on and claims priority to the application with Chinese Application No. 202410903021.8, filed on July 5, 2024, the disclosure of which is hereby incorporated by reference in its entirety into the present disclosure. TECHNICAL FIELD

[0003] The present disclosure relates to the field of in vitro diagnostics, in particular to a sample analyzer for analyzing urine samples, a system for obtaining a kidney damage marker parameter, and a method for analyzing a urine sample. BACKGROUND

[0004] Protein is one of the most important items in urine chemical component examination. Under normal circumstances, due to the pore size barrier and charge barrier of the glomerular filtration membrane, the albumin and globulin of medium and high relative molecular mass in plasma cannot pass through the glomerular filtration membrane; while the proteins with small relative molecular mass, such as β2 microglobulin, α1 microglobulin, lysozyme, etc., can freely pass through the glomerular filtration membrane, but their filtration volume is low, and 95% of them are reabsorbed in the proximal tubule. Therefore, the protein content in the final urine is very small, only 30-130 mg / 24h. The protein in random urine is 0-80 mg / L, and the urine protein qualitative test is negative. When the protein in urine exceeds 150 mg / 24h (or exceeds 100 mg / L), the protein qualitative test of the urine is positive, which is called proteinuria.

[0005] Pathological proteinuria is further divided into glomerular proteinuria and tubular proteinuria.

[0006] Glomerular proteinuria is the most common type of pathological proteinuria under pathological conditions. The glomerulus is damaged by inflammation, toxins and other factors, the charge barrier or (and) pore size barrier of the glomerular filtration membrane is abnormal, and the permeability is increased, so that more medium and large molecular weight plasma proteins are filtered out, which exceeds the reabsorption capacity of the renal tubule and forms proteinuria. The urinary protein excretion of this type of proteinuria can reach more than 2g / 24h, and the urinary protein molecular weight is mostly 70-100kD, mainly albumin, accounting for about 70%-80%. Diseases related to this type of proteinuria include nephrotic syndrome, minimal change nephritis, focal glomerulosclerosis (early stage), membranoproliferative glomerulonephritis (early stage), proliferative glomerulonephritis, diabetic nephropathy stage IV, etc.

[0007] Tubular proteinuria: when the function of renal tubule is impaired, the reabsorption capacity of renal tubule to protein decreases, which can cause the loss of low molecular weight protein. The protein quantification in urine of this type of proteinuria is not more than 1-1.5 g / 24 h, the molecular weight of urine protein is mostly 10-40 kD, and the main components are lysozyme, β2 microglobulin, etc.; if the protein quantification is >2.0 g / 24 h and the molecular weight is >70 kD, glomerular proteinuria should be considered. Diseases related to this type of proteinuria include interstitial nephritis, pyelonephritis, post-transplant rejection, and renal tubular toxicity, etc.

[0008] The total urine protein quantification is located in the upstream position in the diagnosis path of kidney diseases, involves more clinical application scenarios, and has poor specificity. When the total urine protein quantification is abnormal, the conventional method cannot reflect the source of urine protein, and the clinical diagnosis information provided is limited, and usually other test indexes need to be combined to judge the disease condition. Especially, some cases with relatively low total urine protein quantification but with renal tubular damage are missed, which delays the treatment opportunity. SUMMARY

[0009] In order to at least partially solve the above technical problems, the task of the present disclosure is to provide a sample analyzer for analyzing a urine sample, a system for obtaining a kidney damage marker parameter, and an analysis method of a urine sample, which can comprehensively analyze multiple total urine protein quantification results to obtain more valuable clinical information, and especially can help analyze the kidney damage state of a patient.

[0010] In order to achieve the above task of the present disclosure, the first aspect of the present disclosure provides a sample analyzer for analyzing a urine sample, comprising:

[0011] A sample preparation device, comprising a sample carrying part, a sample dispensing part, a reagent carrying part, a reagent dispensing part, and a reaction part, the sample carrying part is used to carry a sample container, the sample container contains a to-be-tested urine sample of a subject, the sample dispensing part is used to suck the to-be-tested urine sample of the subject from the sample carrying part and discharge it into a reaction container to be added, the reagent carrying part is used to carry a detection reagent, the detection reagent includes a colorimetric reagent and a turbidimetric reagent, the reagent dispensing part is used to suck the detection reagent from the reagent carrying part and discharge it into a reaction container to be added with the reagent, and the reaction part is used to place the reaction container so as to incubate a determination test solution obtained by the reaction of the to-be-tested urine sample and the detection reagent in the reaction container, the determination test solution includes a first determination test solution and a second determination test solution, the first determination test solution is obtained by the reaction of the to-be-tested urine sample and the colorimetric reagent, and the second determination test solution is obtained by the reaction of the to-be-tested urine sample and the turbidimetric reagent;

[0012] An optical detection device, comprising a detector for detecting the light signal of the determination test solution; and

[0013] a controller configured to:

[0014] control the sample preparation device to prepare the first assay solution, and control the light detector to detect the first assay solution, so as to obtain a first urine total protein detection result based on a light signal of the first assay solution;

[0015] control the sample preparation device to prepare the second assay solution, and control the light detector to detect the second assay solution, so as to obtain a second urine total protein detection result based on a light signal of the second assay solution;

[0016] in a case where the urine sample to be tested is proteinuria, calculate a kidney injury marker parameter based on the first urine total protein detection result and the second urine total protein detection result, and output the kidney injury marker parameter; and

[0017] when the kidney injury marker parameter is within a first threshold range, it is prompted that the kidney of the subject may be in a clinical pathological state of injury.

[0018] To achieve the above-mentioned tasks of the present disclosure, the second aspect of the present disclosure further provides a system for obtaining a kidney injury marker parameter, comprising:

[0019] a first urine detection module for performing colorimetric determination and turbidimetric determination on a urine sample to be tested of a subject respectively, so as to obtain a first urine total protein detection result based on colorimetric determination and a second urine total protein detection result based on turbidimetric determination; and

[0020] a data analysis module for obtaining the first urine total protein detection result and the second urine total protein detection result; in a case where the urine sample to be tested is proteinuria, calculating a kidney injury marker parameter based on the first urine total protein detection result and the second urine total protein detection result, wherein the kidney injury marker parameter reflects the difference between the first urine total protein detection result and the second urine total protein detection result; and outputting the kidney injury marker parameter.

[0021] To achieve the above-mentioned tasks of the present disclosure, the third aspect of the present disclosure further provides an analysis method of a urine sample, comprising:

[0022] obtaining a first urine total protein detection result of a subject and a second urine total protein detection result of the subject, wherein the first urine total protein detection result is obtained by performing colorimetric determination on a urine sample to be tested of the subject, and the second urine total protein detection result is obtained by performing turbidimetric determination on the urine sample to be tested;

[0023] In a case where the to-be-tested urine sample is a proteinuria sample, a kidney injury marker parameter is calculated according to the first urine total protein detection result and the second urine total protein detection result, wherein the kidney injury marker parameter reflects the difference between the first urine total protein detection result and the second urine total protein detection result; and

[0024] The kidney injury marker parameter is outputted.

[0025] To achieve the above-mentioned tasks of the present disclosure, the fourth aspect of the present disclosure further provides a computer-readable storage medium, wherein a computer program is stored on the computer-readable storage medium, and the computer program is executed by a processor to implement the method according to the third aspect of the present disclosure.

[0026] To achieve the above-mentioned tasks of the present disclosure, the fifth aspect of the present disclosure further provides a sample analyzer for analyzing a urine sample, comprising:

[0027] A sample preparation device, comprising a sample bearing portion, a sample dispensing portion, a reagent bearing portion, a reagent dispensing portion, and a reaction portion, the sample bearing portion is configured to bear a sample container, the sample container contains a to-be-tested urine sample of a subject, the sample dispensing portion is configured to suck the to-be-tested urine sample of the subject from the sample bearing portion and discharge it into a reaction container to be added, the reagent bearing portion is configured to bear a detection reagent, the detection reagent comprises a colorimetric reagent and a turbidimetric reagent, the reagent dispensing portion is configured to suck the detection reagent from the reagent bearing portion and discharge it into a reaction container to be added with the reagent, and the reaction portion is configured to place a reaction container, so as to incubate a determination test solution obtained by the reaction of the to-be-tested urine sample and the detection reagent in the reaction container, the determination test solution comprises a first determination test solution and a second determination test solution, the first determination test solution is obtained by the reaction of the to-be-tested urine sample and the colorimetric reagent, and the second determination test solution is obtained by the reaction of the to-be-tested urine sample and the turbidimetric reagent;

[0028] An optical detection device, comprising an optical detector configured to detect an optical signal of the determination test solution; and

[0029] A controller configured to:

[0030] Control the sample preparation device to prepare the first determination test solution, and control the optical detector to detect the first determination test solution, so as to obtain a first urine total protein detection result based on an optical signal of the first determination test solution;

[0031] Control the sample preparation device to prepare the second determination test solution, and control the optical detector to detect the second determination test solution, so as to obtain a second urine total protein detection result based on an optical signal of the second determination test solution; and

[0032] outputting the first urine total protein test result and the second urine total protein test result.

[0033] In the technical solution provided in the aspects of the present disclosure, the first urine total protein test result based on the colorimetric method and the second urine total protein test result based on the turbidimetric method are comprehensively used to calculate a kidney injury marker parameter, which can reflect the difference between the first urine total protein test result and the second urine total protein test result, thereby being used to evaluate the kidney injury state of a subject.

[0034] The technical solution of the present disclosure will be further described in detail below with the aid of the accompanying drawings and embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0035] In order to more clearly illustrate the technical solutions in the embodiments of the present disclosure or the prior art, the accompanying drawings required to be used in the embodiments or prior art description will be briefly introduced. Obviously, the accompanying drawings in the following description are only some embodiments of the present disclosure, and other accompanying drawings can be obtained by those skilled in the art without any creative effort on the basis of these accompanying drawings.

[0036] FIG. 1 is a structural schematic diagram of a sample analyzer according to some embodiments of the present disclosure.

[0037] FIG. 2 is a structural schematic diagram of a system for obtaining a kidney injury marker parameter according to some embodiments of the present disclosure.

[0038] FIG. 3 is a flow schematic diagram of an analysis method of a urine sample according to some embodiments of the present disclosure.

[0039] FIG. 4 is a schematic diagram of a standard curve of reaction degree and concentration according to some embodiments of the present disclosure.

[0040] FIG. 5 is a schematic diagram of a standard curve of reaction degree and concentration according to some other embodiments of the present disclosure.

[0041] FIG. 6 is a schematic diagram of a relative deviation result according to some embodiments of the present disclosure.

[0042] FIG. 7 is a schematic diagram of a ROC curve according to some embodiments of the present disclosure.

[0043] FIG. 8 is a schematic diagram of a relative deviation result according to some other embodiments of the present disclosure.

[0044] FIG. 9 is a schematic diagram of a relative deviation result according to some further embodiments of the present disclosure.

[0045] FIG. 10 is a schematic diagram of a ROC curve according to some other embodiments of the present disclosure.

[0046] FIG. 11 is a schematic diagram of a ROC curve according to some further embodiments of the present disclosure.

[0047] FIG. 12 is a schematic diagram of results of total urine protein quantification according to some embodiments of the present disclosure.

[0048] FIG. 13 is a schematic diagram of results of relative bias according to yet some embodiments of the present disclosure.

[0049] FIG. 14 is a schematic diagram of grading of tubular type kidney injury according to some embodiments of the present disclosure. DETAILED DESCRIPTION

[0050] The technical solutions in the embodiments of the present disclosure will be apparently and completely described below with the accompanying drawings in the embodiments of the present disclosure. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, rather than all the embodiments of the present disclosure. Based on the embodiments in the present disclosure, all other embodiments obtained by a person of ordinary skill in the art without creative labor fall within the scope of protection of the present disclosure.

[0051] The relative arrangement, numerical expressions, and numerical values of the components and steps set forth in the embodiments are not limiting the scope of the present disclosure, unless otherwise specifically stated.

[0052] Meanwhile, it should be understood that the sizes of the various parts shown in the drawings are not necessarily drawn to scale in order to emphasize certain features of the embodiments.

[0053] The techniques, methods, and devices known to those of ordinary skill in the relevant art can not be discussed in detail, but should be considered as part of the specification where appropriate.

[0054] In all examples shown and discussed herein, any specific values should be interpreted as merely exemplary and not as a limitation. Thus, other examples of the exemplary embodiments can have different values.

[0055] It should be noted that like reference numerals and letters refer to like items in the following drawings, and thus, once an item is defined in one drawing, it need not be discussed further in subsequent drawings.

[0056] In addition, in the description of the present disclosure, the terms "first", "second", "third", and the like are used only for descriptive purposes, and should not be construed as indicating or implying relative importance and sequence. Similarly, although the operations are depicted in a particular order in the drawings, this should not be understood as requiring the operations to be performed in the particular order shown or in a sequential order, or requiring all of the illustrated operations to be performed to achieve the desired result. In some cases, multi-task processing and parallel processing can be advantageous.

[0057] As those skilled in the art will appreciate, unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art in the field of the disclosure.

[0058] ROC curve (receiver operator characteristic curve): receiver operator characteristic curve, is a curve drawn according to a series of different binary classification methods (threshold), with the true positive rate as the vertical coordinate and the false positive rate as the horizontal coordinate, and ROC_AUC (area under the curve) represents the area enclosed by the ROC curve and the horizontal coordinate axis.

[0059] In the related art, in the automatic biochemical analyzer, there are mainly two methods of colorimetry and turbidimetry for detecting proteins in the sample, such as total protein in urine.

[0060] The basic principle of colorimetry is that the dye (or pigment, chromogen) binds with the protein, such as total protein, to produce a color change under certain conditions, and the protein, such as total protein in urine, is detected by measuring the absorbance of light at a certain wavelength or within a certain wavelength range (or by measuring the intensity of transmitted light) of the solution.

[0061] The basic principle of turbidimetry is that turbidimetry reagent binds with protein, such as total protein, to form turbidity, and protein, such as total protein in urine, is detected by measuring the intensity of transmitted light or scattered light. Turbidimetry includes turbidimetry and nephelometry. In turbidimetry, the intensity of light transmitted through the sample is measured; while in nephelometry, the intensity of scattered light is measured.

[0062] Whether it is colorimetry or turbidimetry, a single test of urine total protein provides limited clinical information. For example, when testing a urine sample using a single method, it can only be determined whether the urine sample is proteinuria, but it cannot be determined what type of proteinuria it is.

[0063] Applicants have found through research that, compared with colorimetry, turbidimetry significantly underestimates the measurement of small molecule proteins (such as alpha 1-microglobulin and beta 2-microglobulin, which are small molecule proteins) due to the low turbidity formed after denaturation of small molecule proteins, and such small molecule proteins are markers of renal tubular dysfunction.

[0064] Based on this, the embodiments of the present disclosure propose a technical solution capable of determining urine total protein based on turbidimetry and colorimetry for the same urine sample to obtain more valuable clinical information related to kidney damage.

[0065] Fig. 1 shows a sample analyzer 100 according to an embodiment of the present disclosure, which is configured as a biochemical analyzer.

[0066] As shown in Fig. 1, the sample analyzer 100 comprises a sample preparation device, an optical detection device 160, and a controller 170. The sample preparation device is configured to prepare a test solution, the optical detection device 160 is configured to perform optical detection on the test solution to obtain an optical signal, and the controller 170 is configured to control the sample preparation device and the optical detection device 160 and process the optical signal.

[0067] As shown in Fig. 1, the sample detection device comprises a sample carrying part 110, a sample dispensing part 120, a reagent carrying part 130, a reagent dispensing part 140, and a reaction part 150.

[0068] The sample carrying part 110 is configured to carry a sample container 10, which contains a urine sample to be tested of a subject. For example, the sample carrying part 110 can be configured as a sample tray, which comprises a plurality of sample sites for placing the sample container 10, and the sample tray can be rotated to dispatch the sample container 10 containing the urine sample to be tested to a corresponding position, for example, to a position for the sample dispensing part 120 to aspirate the urine sample to be tested.

[0069] The sample dispensing part 120 is configured to aspirate the urine sample to be tested of the subject from the sample carrying part 110 and discharge it into a reaction container to be added. For example, the sample dispensing part 120 can comprise a sample needle, which can be moved in two or three dimensions in space by a two or three dimensional driving mechanism, so that the sample needle can be moved to a position for aspirating the urine sample to be tested and to a reaction container to be added, and discharge the aspirated urine sample to be tested into the reaction container.

[0070] The reagent carrying part 130 is configured to carry a detection reagent, for example, a colorimetric reagent for measuring the total protein level of urine and a turbidimetric reagent for measuring the total protein level of urine. For example, the reagent carrying part 130 can be configured as a reagent tray having a disc-like structure, which comprises a plurality of positions for carrying reagent containers, and the reagent carrying part 130 can be rotated to rotate the reagent containers carried thereby to a specific position, for example, a position for the reagent dispensing part 140 to aspirate the reagent. The number of reagent carrying parts 130 can be one or more.

[0071] The reagent dispensing part 140 is configured to aspirate the detection reagent from the reagent carrying part 130 and discharge it into a reaction container to be added with the reagent. For example, the reagent dispensing part 140 can comprise a reagent needle, which can be moved in two or three dimensions in space by a two or three dimensional driving mechanism, so that the reagent needle can be moved to a position for aspirating the reagent and to a reaction container to be added with the reagent, and discharge the aspirated reagent into the reaction container.

[0072] The reaction part 150 is configured to place the reaction container, so as to incubate the determination solution in the reaction container, which is obtained by the reaction of the to-be-tested urine sample and the detection reagent. For example, the determination solution includes a first determination solution and a second determination solution, the first determination solution is obtained by the reaction of the to-be-tested urine sample and a first detection reagent, and the second determination solution is obtained by the reaction of the to-be-tested urine sample and a second detection reagent.

[0073] For example, the reaction part 150 can be configured as a reaction disc having a disc-shaped structure, the reaction disc has one or more placement positions for placing the reaction container, and the reaction disc is capable of rotating and driving the reaction container in the placement position to move, so as to schedule the reaction container in the reaction disc and incubate the determination solution in the reaction container.

[0074] The optical detection device 160 includes an optical detector configured to detect the light signal of the determination solution. For example, the optical detection device 160 is arranged outside the reaction part 150, and the reaction part 150 rotates to drive the reaction container containing the determination solution to move to the optical detection device 160 for detection.

[0075] The controller 170 is configured to control the sample preparation device to prepare the first determination solution, i.e., to prepare the first determination solution from the to-be-tested urine sample and the first detection reagent, and to control the optical detector to detect the first determination solution, so as to obtain the first urine total protein detection result based on the light signal of the first determination solution; and to control the sample preparation device to prepare the second determination solution, i.e., to prepare the second determination solution from the to-be-tested urine sample and the second detection reagent, and to control the optical detector to detect the second determination solution, so as to obtain the second urine total protein detection result based on the light signal of the second determination solution. In this application, the optical detector can refer to a single component, or a combination of multiple components forming a set of components capable of detecting the signal of the determination solution.

[0076] Therefore, the sample analyzer 100 provided by the embodiments of the present disclosure can obtain the first urine total protein detection result and the second urine total protein detection result for the same to-be-tested urine sample.

[0077] In the embodiments of the present disclosure, the first urine total protein detection result and the second urine total protein detection result are both urine total protein quantitative detection results.

[0078] In the embodiments of the present disclosure, the reaction principle of the reaction of the first detection reagent with the protein in the urine sample is different from the reaction principle of the reaction of the second detection reagent with the protein in the urine sample.

[0079] In some embodiments, the first detection reagent is a colorimetric reagent, and the second detection reagent is a turbidimetric reagent.

[0080] In one example, a specific detection procedure for detecting total urine protein in a urine sample to be tested using the sample analyzer 100 is as follows: the sample dispensing unit 120 sucks the urine sample to be tested from the sample carrier 110 and adds it to a reaction container, the light detector of the optical detection device 160 records the absorbance of the urine sample to be tested at this time (denoted as A1); the reagent dispensing unit 140 sucks the colorimetric reagent (or turbidimetric reagent) from the reagent carrier 130 and adds it to the reaction container to which the urine sample to be tested has been added, so as to mix with the urine sample to be tested; then the reaction container is placed in the reaction unit 150 for reaction and incubation to obtain a determination solution; then the light detector of the optical detection device 160 records the absorbance of the determination solution at this time (denoted as A2), so as to calculate the concentration level of total urine protein by the measured A1 and A2.

[0081] In some embodiments, the controller 170 is further configured to output the first total urine protein detection result and the second total urine protein detection result.

[0082] In some alternative or additional embodiments, the controller 170 is further configured to, in the case that the urine sample to be tested is proteinuria, calculate a kidney injury marker parameter for characterizing the kidney injury state of the subject according to the first total urine protein detection result and the second total urine protein detection result; and output the kidney injury marker parameter.

[0083] In this way, the kidney injury marker parameter for characterizing the kidney injury state of the subject can be obtained based on the difference between the first total urine protein detection result and the second total urine protein detection result, thereby helping to judge different types of proteinuria.

[0084] Here, the kidney injury marker parameter obtained by the embodiments of the present disclosure can be used to evaluate the kidney injury state of the subject, for example, can be used to prompt whether the kidney of the subject is in a clinical pathological state of injury, to identify different types of kidney injury, to predict the risk of kidney injury, and to manage the risk grading of kidney injury.

[0085] In some embodiments, the optical detection device 160 comprises a transmittance light detector for detecting the transmittance light signal of the first assay solution and the second assay solution. Accordingly, the controller 170 controls the light detector to detect the first assay solution so as to obtain the first urine total protein test result based on the light signal of the first assay solution, including: the controller 170 controls the transmittance light detector to detect the first assay solution so as to obtain the first urine total protein test result based on the transmittance light signal of the first assay solution. The controller 170 controls the light detector to detect the second assay solution so as to obtain the second urine total protein test result based on the light signal of the second assay solution, including: the controller 170 controls the transmittance light detector to detect the second assay solution so as to obtain the second urine total protein test result based on the transmittance light signal of the second assay solution. That is, the same transmittance light detector is used to perform transmittance light assay on the first assay solution and the second assay solution.

[0086] In other embodiments, the optical detection device 160 comprises a transmittance light detector for detecting the transmittance light signal of the first assay solution and a scattering light detector for detecting the scattering light signal of the second assay solution. Accordingly, the controller 170 controls the light detector to detect the first assay solution so as to obtain the first urine total protein test result based on the light signal of the first assay solution, including: the controller 170 controls the transmittance light detector to detect the first assay solution so as to obtain the first urine total protein test result based on the transmittance light signal of the first assay solution. The controller 170 controls the light detector to detect the second assay solution so as to obtain the second urine total protein test result based on the light signal of the second assay solution, including: the controller 170 controls the scattering light detector to detect the second assay solution so as to obtain the second urine total protein test result based on the scattering light signal of the second assay solution. That is, different light detectors are used to perform light assay on the first assay solution and the second assay solution, respectively.

[0087] In some embodiments, the controller 170 is further configured to: when the kidney injury marker parameter is within the first threshold range, then prompt that the kidney of the subject is likely to be in a clinical pathologic state of injury.

[0088] In some embodiments, the controller 170 is further configured to: differentiate glomerular type of kidney injury and tubular type of kidney injury of the subject according to the kidney injury marker parameter. That is, whether the subject has glomerular type of kidney injury or tubular type of kidney injury can be differentiated by the kidney injury marker parameter.

[0089] As some implementations, the kidney injury marker parameter is used to differentiate between glomerular and tubular kidney injury in the subject, including: identifying the subject as having glomerular kidney injury when the kidney injury marker parameter is within a second threshold range; and identifying the subject as having tubular kidney injury when the kidney injury marker parameter is outside the second threshold range.

[0090] For example, the subject is identified as having glomerular kidney injury when the kidney injury marker parameter is greater than a preset threshold; and the subject is identified as having tubular kidney injury when the kidney injury marker parameter is less than the preset threshold.

[0091] That is, in the case of judging the urine sample of the subject as proteinuria, especially pathological proteinuria, if the value of the kidney injury marker parameter is greater than a cutoff value, the subject is judged as having glomerular kidney injury, and vice versa.

[0092] In some embodiments, the controller 170 is further configured to predict the risk of tubular kidney injury in the subject according to the kidney injury marker parameter, and output the corresponding prediction result. That is, whether the subject has the risk of tubular kidney injury can be predicted by the kidney injury marker parameter.

[0093] For example, the controller 170 is further configured to predict that the subject has the risk of tubular kidney injury when the kidney injury marker parameter is within a preset threshold range.

[0094] In some embodiments, the controller 170 is further configured to classify the risk of tubular kidney injury in the subject according to the kidney injury marker parameter, and output the risk classification. In particular, the controller 170 is further configured to classify the risk of tubular kidney injury in the subject according to the kidney injury marker parameter, in the case of predicting that the subject has the risk of tubular kidney injury according to the kidney injury marker parameter.

[0095] As some implementations, the controller 170 is further configured to classify the risk of tubular kidney injury in the subject into low risk, medium risk and high risk according to the kidney injury marker parameter.

[0096] It should be understood that low risk means that the subject has a low possibility of having tubular kidney injury, medium risk means that the subject has a moderate possibility of having tubular kidney injury (i.e., the possibility of the subject having tubular kidney injury in the case of medium risk is greater than that in the case of low risk), and high risk means that the subject has a high possibility of having tubular kidney injury (i.e., the possibility of the subject having tubular kidney injury in the case of high risk is greater than that in the case of medium risk).

[0097] As some implementations, classifying the risk of tubular kidney injury of the subject according to the kidney injury marker parameter as low risk, medium risk and high risk includes: outputting a prompt indicating that the tubular kidney injury of the subject is low risk when the kidney injury marker parameter is within a third threshold range; outputting a prompt indicating that the tubular kidney injury of the subject is medium risk when the kidney injury marker parameter is within a fourth threshold range and outside the third threshold range; and outputting a prompt indicating that the tubular kidney injury of the subject is high risk when the kidney injury marker parameter is within a fifth threshold range and outside the third threshold range and the fourth threshold range.

[0098] For example, outputting a prompt indicating that the tubular kidney injury of the subject is low risk when the kidney injury marker parameter is greater than a first threshold value; outputting a prompt indicating that the tubular kidney injury of the subject is medium risk when the kidney injury marker parameter is less than or equal to the first threshold value and greater than or equal to a second threshold value; and outputting a prompt indicating that the tubular kidney injury of the subject is high risk when the kidney injury marker parameter is less than the second threshold value.

[0099] In some embodiments, the controller 170 is further configured to determine whether the urine sample of the subject is proteinuria according to the first total urine protein detection result; or receive input information and determine whether the urine sample of the subject is proteinuria based on the input information.

[0100] As some implementations, the input information includes urine routine information of the subject.

[0101] As other implementations, the input information includes department information of the subject.

[0102] As some implementations, the user can input the input information (e.g., urine routine information) through a human-computer interaction interface, or the sample analyzer can read the input information from a storage medium, or the sample analyzer can obtain the urine routine information from a urine routine detection device, such as a urine analyzer based on dry chemical test paper or a urine sediment analyzer. It should be understood that the manner of obtaining the input information is not limited thereto.

[0103] In some embodiments, the controller 170 is further configured to prepare the first assay solution and the second assay solution by:

[0104] controlling the sample preparation device to mix a portion of the urine sample to be tested with a first detection reagent, such as a colorimetric reagent, to prepare the first assay solution; and controlling the sample preparation device to mix at least a portion of the first assay solution with a second detection reagent, such as a turbidimetric reagent, to prepare the second assay solution.

[0105] That is, the first assay test solution can be prepared first, and then a portion of the first assay test solution is mixed with the second detection reagent, e.g. the turbidimetric reagent, to prepare the second assay test solution.

[0106] In one specific example, the controller 170 is further configured to prepare the first assay test solution by controlling the sample dispensing portion 120 to aspirate the urine sample to be measured from the sample container in the sample carrier portion 110 and add it to the first reaction container, controlling the reagent dispensing portion 140 to aspirate the colorimetric reagent from the reagent carrier portion 130 and add it to the first reaction container to which the urine sample to be measured has been added, so as to mix with the urine sample to be measured, and controlling the first reaction container to be placed in the reaction portion 150 for reaction and incubation to obtain the first assay test solution. Correspondingly, the controller 170 is further configured to prepare the second assay test solution by controlling the reagent dispensing portion 140 to aspirate the turbidimetric reagent from the reagent carrier portion 130 and add it to the first reaction container, so as to mix with the first assay test solution to obtain the second assay test solution, or controlling the sample dispensing portion 120 to aspirate a portion of the first assay test solution from the first reaction container and add it to the second reaction container, controlling the reagent dispensing portion 140 to aspirate the turbidimetric reagent from the reagent carrier portion 130 and add it to the second reaction container to which the portion of the first assay test solution has been added, so as to mix with the portion of the first assay test solution, and controlling the second reaction container to be placed in the reaction portion 150 for reaction and incubation to obtain the second assay test solution.

[0107] In other embodiments, the controller 170 is further configured to prepare the first assay test solution and the second assay test solution by:

[0108] controlling the sample preparation device to mix a first portion of the urine sample to be measured with a first detection reagent, e.g. the colorimetric reagent, to prepare the first assay test solution, and controlling the sample preparation device to mix a second portion of the urine sample to be measured with a second detection reagent, e.g. the turbidimetric reagent, to prepare the second assay test solution.

[0109] It should be understood that the first portion and the second portion are non-overlapping portions.

[0110] In one specific example, the controller 170 is further configured to prepare the first assay test solution by: controlling the sample dispensing portion 120 to draw a first portion of the urine sample under test from the sample container in the sample carrier 110 and add it to the first reaction container; controlling the reagent dispensing portion 140 to draw the colorimetric reagent from the reagent carrier 130 and add it to the first reaction container to which the first portion of the urine sample under test has been added, so as to mix with the first portion of the urine sample under test; and controlling the first reaction container to be placed into the reaction portion 150 for reaction, incubation to obtain the first assay test solution. Correspondingly, the controller 170 is further configured to prepare the second assay test solution by: controlling the sample dispensing portion 120 to draw a second portion of the urine sample under test from the sample container and add it to the second reaction container; controlling the reagent dispensing portion 140 to draw the turbidimetric reagent from the reagent carrier 130 and add it to the second reaction container to which the second portion of the urine sample under test has been added, so as to mix with the second portion of the urine sample under test; and controlling the second reaction container to be placed into the reaction portion 150 for reaction, incubation to obtain the second assay test solution.

[0111] In some embodiments, the controller 170 is further configured to calculate the kidney injury marker parameter according to a first algorithm or according to a second algorithm.

[0112] Here, the first algorithm includes any one of a relative deviation, an absolute deviation, a relative mean deviation and a weighting of the first total urine protein detection result and the second total urine protein detection result, and the second algorithm includes any one of a relative deviation, an absolute deviation and a weighting of a logarithm of the first total urine protein detection result and a logarithm of the second total urine protein detection result.

[0113] In some embodiments, the calculation algorithm is selected so as to eliminate interference, such as interference of total protein concentration, generated in the process of comparing the first total urine protein detection result and the second total urine protein detection result, for example by highlighting a deviation of the first total urine protein detection result from the second total urine protein detection result.

[0114] As some implementations, the controller 170 is further configured to calculate the kidney injury marker parameter by the calculation formula of Table 1, wherein C1 is the first total urine protein detection result and C2 is the second total urine protein detection result.

[0115] Table 1

[0116] In some embodiments, the reagent carrier 130 is configured to place a reagent storage device. Here, the first detection reagent, such as the colorimetric reagent, and the second detection reagent, such as the turbidimetric reagent, are stored in the same reagent storage device. For example, the first detection reagent and the second detection reagent are stored in the same reagent cartridge.

[0117] In some embodiments, the first detection reagent is a colorimetric reagent, including a chromogenic reagent. A chromogenic reagent refers to a substance capable of reacting with a protein to produce a change in spectrum.

[0118] In some embodiments, the colorimetric protein quantification method includes the ortho-phenanthroline red protein quantification method, the bicinchoninic acid (BCA) protein quantification method, the biuret protein quantification method, and the Lowery protein quantification method, etc.

[0119] In some embodiments, the second detection reagent is a turbidimetric reagent, for example including a protein precipitation reagent. A protein precipitation reagent refers to a substance capable of precipitating a protein to produce turbidity. For example, the protein precipitation reagent is sulfosalicylic acid or trichloroacetic acid, etc.

[0120] In some embodiments, the turbidimetric protein quantification method includes the benzethonium chloride protein quantification method, the benzalkonium chloride protein quantification method, and the sulfosalicylic acid protein quantification method, etc.

[0121] In some embodiments, the same type of protein quantification method also has different affinities for different proteins due to different reagent formulations. For example, adding a specific surfactant in the ortho-phenanthroline red protein quantification method can improve its affinity for globulin.

[0122] Of course, the present disclosure is not limited thereto, and in theory, the combination of two different total protein quantification methods falls within the protection scope of the present disclosure.

[0123] In some embodiments, the controller 170 is further configured to: control the sample preparation device to prepare the first assay reagent and control the light detector to detect the first assay reagent to obtain the first urine total protein detection result, and control the sample preparation device to prepare the second assay reagent and control the light detector to detect the second assay reagent to obtain the second urine total protein detection result, regardless of whether the urine sample to be tested is proteinuria.

[0124] That is, in these embodiments, for any urine sample of a subject, whether it is proteinuria or not, the urine sample is subjected to colorimetric determination and turbidimetric determination to obtain the first urine total protein detection result and the second urine total protein detection result. And only in the case of judging that the urine sample of the subject is proteinuria, the kidney injury marker parameter is calculated based on the first urine total protein detection result and the second urine total protein detection result.

[0125] In other embodiments, the controller 170 is further configured to: control the sample preparation device to prepare the first assay reagent and control the light detector to detect the first assay reagent to obtain the first urine total protein detection result, and control the sample preparation device to prepare the second assay reagent and control the light detector to detect the second assay reagent to obtain the second urine total protein detection result, only in the case of judging that the urine sample to be tested is proteinuria.

[0126] For example, whether the urine sample to be tested is proteinuria can be determined by the input information in the foregoing embodiments, such as the urine routine information of the subject.

[0127] That is, in the case where the urine sample of the subject is determined to be proteinuria, the colorimetric determination and the turbidimetric determination are performed on the urine sample to obtain the first urine total protein detection result and the second urine total protein detection result, and then the kidney injury marker parameter is calculated based on the first urine total protein detection result and the second urine total protein detection result.

[0128] In some embodiments, the controller 170 is further configured to: control the sample preparation device to prepare the first determination reagent and control the light detector to detect the first determination reagent to obtain the first urine total protein detection result; and control the sample preparation device to prepare the second determination reagent and control the light detector to detect the second determination reagent to obtain the second urine total protein detection result only in the case where the urine sample to be tested is determined to be proteinuria.

[0129] Here, preferably, whether the urine sample to be tested is proteinuria is determined according to the first urine total protein detection result. Alternatively, whether the urine sample to be tested is proteinuria can be determined by the input information in the foregoing embodiments, such as the urine routine information of the subject.

[0130] That is, in these embodiments, for any urine sample of a subject, whether it is proteinuria or not, the colorimetric determination is performed on the urine sample to obtain the first urine total protein detection result. In the case where the urine sample of the subject is determined to be proteinuria (for example, according to the first urine total protein detection result), the turbidimetric determination is performed on the urine sample to obtain the second urine total protein detection result, and then the kidney injury marker parameter is calculated based on the first urine total protein detection result and the second urine total protein detection result.

[0131] The present disclosure also proposes a system for obtaining a kidney injury marker parameter.

[0132] FIG. 2 is a structural schematic diagram of a system 200 for obtaining a kidney injury marker parameter according to some embodiments of the present disclosure.

[0133] As shown in FIG. 2, the system 200 for obtaining a kidney injury marker parameter includes a first urine detection module 201 and a data analysis module 202.

[0134] The first urine detection module 201 is configured to perform colorimetric determination and turbidimetric determination on a urine sample to be tested of a subject, respectively, to obtain a first urine total protein detection result based on the colorimetric determination and a second urine total protein detection result based on the turbidimetric determination.

[0135] The data analysis module 202 is configured to obtain the first urine total protein test result and the second urine total protein test result; in the case that the to-be-tested urine sample is proteinuria, calculate a kidney injury marker parameter according to the first urine total protein test result and the second urine total protein test result; and output the kidney injury marker parameter. Here, the kidney injury marker parameter reflects the difference between the first urine total protein test result and the second urine total protein test result.

[0136] In some embodiments, the data analysis module 202 is further configured to: when the kidney injury marker parameter is within a first threshold range, prompt that the subject's kidney is likely to be in a clinical pathological state of injury; and / or identify glomerular type kidney injury and tubular type kidney injury of the subject according to the kidney injury marker parameter; and / or predict the risk of tubular type kidney injury of the subject according to the kidney injury marker parameter, and output the corresponding prediction result; and / or risk stratify the tubular type kidney injury of the subject according to the kidney injury marker parameter, and output the risk stratification.

[0137] In some embodiments, identifying glomerular type kidney injury and tubular type kidney injury of the subject according to the kidney injury marker parameter comprises: when the kidney injury marker parameter is within a second threshold range, identifying the subject as having glomerular type kidney injury; and when the kidney injury marker parameter is outside the second threshold range, identifying the subject as having tubular type kidney injury.

[0138] For example, when the kidney injury marker parameter is greater than a preset threshold, identifying the subject as having glomerular type kidney injury; and when the kidney injury marker parameter is less than the preset threshold, identifying the subject as having tubular type kidney injury.

[0139] Alternatively or additionally, predicting the risk of tubular type kidney injury of the subject according to the kidney injury marker parameter, and outputting the corresponding prediction result, comprises: when the kidney injury marker parameter is within a preset threshold range, predicting that the subject has a risk of tubular type kidney injury.

[0140] Alternatively or additionally, risk stratifying the tubular type kidney injury of the subject according to the kidney injury marker parameter, and outputting the risk stratification, comprises: when the kidney injury marker parameter is within a third threshold range, outputting a prompt indicating that the tubular type kidney injury of the subject is low risk; when the kidney injury marker parameter is within a fourth threshold range and outside the third threshold range, outputting a prompt indicating that the tubular type kidney injury of the subject is medium risk; and when the kidney injury marker parameter is within a fifth threshold range and outside the third threshold range and the fourth threshold range, outputting a prompt indicating that the tubular type kidney injury of the subject is high risk.

[0141] For example, when the kidney injury marker parameter is greater than the first threshold value, a prompt indicating that the tubular type kidney injury of the subject is low risk is output; when the kidney injury marker parameter is less than or equal to the first threshold value and greater than or equal to the second threshold value, a prompt indicating that the tubular type kidney injury of the subject is medium risk is output; and when the kidney injury marker parameter is less than the second threshold value, a prompt indicating that the tubular type kidney injury of the subject is high risk is output.

[0142] In some embodiments, the data analysis module 202 is further configured to determine whether the urine sample of the subject is proteinuria according to the first urine total protein detection result.

[0143] In some embodiments, the data analysis module 202 is further configured to receive input information and determine whether the urine sample of the subject is proteinuria based on the input information.

[0144] As some implementations, the system 200 for obtaining the kidney injury marker parameter further comprises a second urine detection device for obtaining urine routine information of the subject, and the data analysis module 202 is further configured to obtain the urine routine information and determine whether the urine sample of the subject is proteinuria based on the urine routine information.

[0145] In some embodiments, the first urine detection module 210 comprises a biochemical analyzer, for example, the sample analyzer 100 according to the above embodiments, and the data analysis module 202 is integrated in the controller of the biochemical analyzer in the form of software or hardware or integrated in the computer connected with the biochemical analyzer.

[0146] In some embodiments, the second urine detection device comprises at least one of a urine analyzer based on dry chemical test paper method and a urine sediment analyzer.

[0147] The present disclosure also proposes an analysis method of a urine sample.

[0148] FIG. 3 is a flow diagram of an analysis method 300 of a urine sample according to some embodiments of the present disclosure.

[0149] In step S302, a first urine total protein detection result of the subject and a second urine total protein detection result of the subject are obtained. Here, the first urine total protein detection result is obtained by colorimetric determination on the to-be-tested urine sample of the subject, and the second urine total protein detection result is obtained by turbidimetric determination on the to-be-tested urine sample.

[0150] In step 304, in the case that the to-be-tested urine sample is proteinuria, a kidney injury marker parameter is calculated according to the first urine total protein detection result and the second urine total protein detection result. Here, the kidney injury marker parameter reflects the difference between the first urine total protein detection result and the second urine total protein detection result.

[0151] At step 306, outputting the renal injury marker parameter.

[0152] In some embodiments, the analysis method 300 further comprises: when the renal injury marker parameter is within a first threshold range, then prompting that the subject’s kidney is likely in a clinical pathologic state of injury; and / or, differentiating glomerular-type renal injury and tubular-type renal injury of the subject according to the renal injury marker parameter; and / or, predicting the risk of tubular-type renal injury of the subject according to the renal injury marker parameter, and outputting a corresponding prediction result; and / or, risk stratifying the tubular-type renal injury of the subject according to the renal injury marker parameter, and outputting a risk stratification.

[0153] In some embodiments, differentiating glomerular-type renal injury and tubular-type renal injury of the subject according to the renal injury marker parameter comprises: when the renal injury marker parameter is within a second threshold range, differentiating the subject as having glomerular-type renal injury; and / or, when the renal injury marker parameter is outside the second threshold range, differentiating the subject as having tubular-type renal injury.

[0154] For example, when the renal injury marker parameter is greater than a preset threshold, differentiating the subject as having glomerular-type renal injury; and / or, when the renal injury marker parameter is less than the preset threshold, differentiating the subject as having tubular-type renal injury.

[0155] Alternatively or additionally, predicting the risk of tubular-type renal injury of the subject according to the renal injury marker parameter, and outputting a corresponding prediction result, comprises: when the renal injury marker parameter is within a preset threshold range, predicting that the subject has a risk of tubular-type renal injury.

[0156] Alternatively or additionally, risk stratifying the tubular-type renal injury of the subject according to the renal injury marker parameter, and outputting a risk stratification, comprises: when the renal injury marker parameter is within a third threshold range, outputting a prompt indicating that the tubular-type renal injury of the subject is at a low risk; when the renal injury marker parameter is within a fourth threshold range and outside the third threshold range, outputting a prompt indicating that the tubular-type renal injury of the subject is at a medium risk; and / or, when the renal injury marker parameter is within a fifth threshold range and outside the third threshold range and the fourth threshold range, outputting a prompt indicating that the tubular-type renal injury of the subject is at a high risk.

[0157] For example, when the renal injury marker parameter is greater than a first threshold, outputting a prompt indicating that the tubular-type renal injury of the subject is at a low risk; when the renal injury marker parameter is less than or equal to the first threshold and greater than or equal to a second threshold, outputting a prompt indicating that the tubular-type renal injury of the subject is at a medium risk; and / or, when the renal injury marker parameter is less than the second threshold, outputting a prompt indicating that the tubular-type renal injury of the subject is at a high risk.

[0158] The embodiment of the present disclosure further provides a computer readable storage medium comprising computer program instructions, which, when executed by a processor, implement the method of any one of the above-mentioned embodiments.

[0159] Those skilled in the art should understand that each module or each step of the embodiment of the present disclosure can be implemented by a general computing device. The general computing device generally comprises a processor and a memory for storing instructions, which, when executed by the processor, cause the computing device to perform each step or each program module of the embodiment of the present disclosure.

[0160] In the embodiment of the present disclosure, the memory is used for storing computer programs or instructions, and the memory can be: a volatile memory such as a random access memory (RAM), or a non-volatile memory such as a read only memory (ROM), a flash memory, a hard disk drive (HDD) or a solid state drive (SSD), or a combination of the above kinds of memories.

[0161] Those skilled in the art should understand that the embodiment of the present disclosure can be provided as a method, a system or a computer program product. Therefore, the embodiment of the present disclosure can be in the form of a hardware embodiment, a software embodiment or an embodiment combining software and hardware aspects. Moreover, the embodiment of the present disclosure can be in the form of a computer program product implemented on one or more computer usable storage media (including magnetic disk storage and optical storage, etc.) containing computer usable program code.

[0162] It should be understood that the above-mentioned various embodiments of the sample analyzer 100 proposed by the present disclosure and the advantages thereof are also applicable to the system 200, the method 300 and the computer readable storage medium proposed by the present disclosure.

[0163] The technical solutions of the present disclosure are verified by means of some embodiments as follows.

[0164] Embodiment one: Differentiation of glomerular injury and urinary tubular injury based on self-prepared reagent

[0165] 1. Collecting urine samples and patient information of patients and screening into samples

[0166] 1) Patient information: gender, age, department, diagnosis information, and total urine protein quantitative measurement value

[0167] 2) Screening of samples: urine samples from patients over 18 years old, urine color is light yellow, and the patients are diagnosed by clinicians to have glomerular or tubular damage, and the urine total protein concentration is between 150-2000 mg / L (i.e. proteinuria).

[0168] The specific case sample conditions are shown in Table 2.

[0169] Table 2

[0170] 2. Test the urine samples by using two total protein quantification methods (i.e. colorimetric method and transmission turbidimetry method) and calculate the kidney damage marker parameters

[0171] 1) Reagent preparation

[0172] Colorimetric reagent A: o-phenanthroline red 27 mg / L sodium molybdate 30 mg / L

[0173] Turbidimetric reagent B: sulfosalicylic acid 30 g / L

[0174] 2) Detection method: add 3 μL of the urine sample to be tested to 100 μL of the colorimetric reagent A to prepare a first test solution, incubate the first test solution at 37°C for 5 minutes, record the change in absorbance at 605 nm before and after adding the urine sample to be tested, the absorbance before adding the sample is A1, and the absorbance after adding the sample is A2; add 100 μL of the turbidimetric reagent B to the first test solution, record the change in absorbance at 380 nm before and after adding the turbidimetric reagent B, the absorbance before adding the reagent is A'1, and the absorbance after adding the reagent is A'2; calculate the reaction degree R1 of the colorimetric reagent A from the change in absorbance: R1 = A2-100 / (100+3) x A1; calculate the reaction degree R2 of the turbidimetric reagent B from the change in absorbance: R2 = A'2-100 / (100+3) x A'1.

[0175] Using the above detection method, a standard curve S1 of the reaction degree R1 and the concentration (see Figure 4) and a standard curve S2 of the reaction degree R2 and the concentration (see Figure 5) are established using albumin. The linear range of the colorimetric reagent A and the turbidimetric reagent B is 100 mg / L to 3200 mg / L. In pathological proteinuria, there are many types of proteins and complex protein composition, and albumin is the most abundant protein in pathological proteinuria, so the urine total protein quantification reagent generally uses albumin to establish a standard curve.

[0176] The enrolled test urine samples were stored at -80°C, and the test urine samples were left to thaw completely at room temperature before detection. The thawed test urine samples were centrifuged at 5000xg for 10 min, and the supernatant was taken for detection. The supernatant was transferred to a disposable collection tube, and each enrolled test urine sample was tested for urine total protein concentration using the Mindray BS-2800 biochemical analyzer according to the above detection method. The first urine total protein detection result (hereinafter referred to as the first urine total protein concentration) was obtained based on the reaction rate R1 and the standard curve S1 of each enrolled sample; the second urine total protein detection result (hereinafter referred to as the second urine total protein concentration) was obtained based on the reaction rate R2 and the standard curve S2 of each enrolled sample; and the relative deviation of the first urine total protein concentration and the second urine total protein concentration was calculated as a kidney injury marker parameter: (second urine total protein concentration / first urine total protein concentration-1) x 100%. The first urine total protein concentration, the second urine total protein concentration and the relative deviation of each enrolled sample are shown in Table 3 and Fig. 6, wherein the urine total protein concentration is in mg / L.

[0177] Table 3

[0178] According to the Welch's t test analysis, a two-tailed test was used without assuming that the standard deviations of the two types of samples are the same. The mean relative deviation of the glomerular injury samples was -34.53%, while that of the tubular injury samples was -61.24%. The P value of the two groups of data was <0.0001, which had a significant difference, as shown in Table 4.

[0179] Table 4

[0180] 3. ROC curve and cutoff value establishment

[0181] The cutoff value was established according to the relative deviation of the first urine total protein concentration and the second urine total protein concentration. The calculation method of the Youden index was: sensitivity + specificity - 1. The Youden index integrated the sensitivity and specificity, and the performance difference caused by different cutoff values was as follows, as shown in Table 5. The maximum Youden index was 0.5356, and the corresponding optimal diagnostic cutoff value was -27.7%. Under this cutoff value, the sensitivity of the method was 91.8%, and the specificity was 61.76%.

[0182] Table 5

[0183] The ROC curve established according to the relative deviation of the first total urine protein concentration and the second total urine protein concentration is shown in FIG. 7, the area under the curve (AUC) is 0.7932, and the P value is less than 0.0001, which has a significant difference. The AUC is between 0.7 and 0.9, indicating that the renal injury marker parameter proposed in the embodiment of the present disclosure has high diagnostic efficiency for the identification of glomerular injury and urinary tubular injury.

[0184] 4. AUC comparison of different algorithms

[0185] The renal injury marker parameters were calculated based on each algorithm of Table 1 mentioned above, and the corresponding ROC curves were established, and the area under the curve (AUC) shown in Table 6 was obtained.

[0186] Table 6

[0187] Generally, the larger the AUC, the better the discrimination ability of the prediction model. In the art, an AUC value greater than 0.75 is considered to have good discrimination ability. In the embodiment of the present disclosure, the AUC of the renal injury marker parameter obtained according to different algorithms is greater than 0.75, which proves that the model according to the embodiment of the present disclosure has good discrimination degree for the population of glomerular injury and the population of urinary tubular injury, indicating that the model has predictive value for the type of renal injury.

[0188] Example Two: Discrimination of glomerular injury and urinary tubular injury prediction based on commercial reagents

[0189] 1. Collect urine samples and patient information of patients and screen the enrolled samples

[0190] The same as Example One

[0191] 2. Test the urine samples using commercial total protein quantification kits and calculate the renal injury marker parameters

[0192] The commercial kits and methodologies used in the experiment are shown in Table 7.

[0193] Table 7

[0194] The enrolled urine samples to be tested were tested according to the instructions using commercial kits on the corresponding biochemical analyzers. When performing the biuret method test, the sample amount was appropriately increased to ensure the accuracy of the test.

[0195] The test results of different methodologies are shown in Table 8, wherein the total protein concentration unit is mg / L.

[0196] Table 8

[0197] The relative deviation of the total urine protein concentration according to different methodologies is calculated as a kidney injury marker parameter, wherein the relative deviation of the benzothiazolium method and the benzethonium chloride method is (the benzethonium chloride measured value / the benzothiazolium measured value-1) x 100%, and the relative deviation of the biuret method and the benzethonium chloride method is (the benzethonium chloride measured value / the biuret measured value-1) x 100%. The relative deviation results of the different method values of the glomerular and tubular injury urine samples are shown in FIGS. 8 and 9.

[0198] According to the Welch's t test analysis, a two-tailed test is used without assuming that the standard deviations of the two types of samples are the same. In the case of comparison of the two methodologies, the P values of the two groups of data are less than 0.0001, which are significantly different, as shown in Table 9.

[0199] Table 9

[0200] 3. ROC curve establishment

[0201] The ROC curve is established in the same manner as in Example 1. According to the relative deviation algorithm, the ROC curves are shown in FIGS. 10 and 11. The area under the curve (Area) of the benzothiazolium method and the benzethonium chloride method is 0.7359, and the P value is less than 0.0001, which is significantly different. The area under the curve (Area) of the biuret method and the benzethonium chloride method is 0.7329, and the P value is less than 0.0001, which is significantly different. The AUC of the kidney injury marker parameters obtained by using the commercial reagents is between 0.7 and 0.9, which indicates that the kidney injury marker parameters proposed in the present disclosure have high diagnostic efficiency for the identification of glomerular injury and urinary tubular injury in the case of using commercial kits of different methodologies.

[0202] Example Three: Tubular injury risk prediction and risk grading

[0203] 1. Collect urine samples and patient information and screen the enrolled samples

[0204] 1) Patient information: gender, age, department

[0205] 2) Screening of enrolled samples: urine samples from patients over 18 years old, urine is light yellow, and the total urine protein concentration is between 150-2000 mg / L.

[0206] 3) 398 cases of final enrolled urine samples

[0207] 2. Test the urine samples by using two total protein quantification methods and calculate the kidney injury marker parameters

[0208] The same as in Example 1

[0209] 3. Urine sample tubular injury biomarker test, and tubular injury prediction and injury grading The urine samples were tested for the presence of a1-MG, b2-MG and RBP (tubular injury biomarkers) using commercial in vitro diagnostic kits, and the total urine protein was quantified using a commercial colorimetric kit. According to the reference range, the items exceeding the reference range were marked as positive, and those within the reference range were marked as negative. If all three tubular injury biomarkers in the urine sample were negative, it was considered to be non-positive, if one tubular injury biomarker was positive, it was considered to be single-positive, if two tubular injury biomarkers were positive, it was considered to be double-positive, and if three tubular injury biomarkers were positive, it was considered to be triple-positive. The results of the total urine protein concentration analysis of the four groups are shown in Table 10 and Figure 12.

[0210] As shown in Table 10 and Figure 12, there was no significant difference in the total urine protein quantification of the four groups of urine samples, indicating that the total urine protein results determined by the existing urine protein kit cannot reflect the degree of tubular injury.

[0211] Table 10

[0212] The present inventors unexpectedly found that the difference between the total urine protein results determined by the colorimetric and turbidimetric methods has clinical relevance to tubular injury.

[0213] The turbidimetric and colorimetric methods were used to determine the concentration of the urine samples, and the relative deviation was calculated.

[0214] The relative deviation calculation formula is: (turbidimetric quantification result / colorimetric quantification result - 1) x 100%.

[0215] The results are shown in Table 11 and Figure 13. There was a significant difference between the non-positive group and the single-positive group (P = 0.001), a significant difference between the single-positive group and the double-positive group (P = 0.0051), and no significant difference between the double-positive group and the triple-positive group (P = 0.9686). This indicates that the difference between the colorimetric and turbidimetric total urine protein results can be used to a certain extent to predict the risk of tubular injury. In addition, there was no significant difference between the double-positive group and the triple-positive group, while there was a significant difference between the non-positive group and the single-positive group, and between the single-positive group and the double-positive group, indicating that the renal injury marker parameter proposed in the present disclosure can be used to a certain extent to predict the risk of early tubular injury.

[0216] Table 11

[0217] In addition, the risk of tubular injury can also be graded according to the relative deviation of the two urine total protein quantification method values. The no-positive group and the single-positive group were compared, and the best separation value of the two groups according to the Youden index was -43.05%; the single-positive group and the double-positive group were compared, and the best separation value of the two groups according to the Youden index was -62.30%, see Table 12. According to the separation value, different urine samples can be divided into three risk levels according to the relative deviation: low risk (relative deviation > -43.05%), medium risk (relative deviation between -43.05% and -62.30%), and high risk (relative deviation < -62.30%).

[0218] Table 12

[0219] By comparing the specific proportion of no-positive, single-positive, double-positive and triple-positive in urine samples of different risk levels, see Figure 14, it can be seen that as the risk level increases, the proportion of no-positive urine samples gradually decreases, and the proportion of triple-positive urine samples gradually increases, which illustrates the effectiveness of using the renal injury marker parameters proposed in the embodiments of the present disclosure for risk grading of tubular injury.

[0220] So far, the embodiments of the present disclosure have been described in detail. In order to avoid obscuring the concept of the present disclosure, some details known in the art are not described. Those skilled in the art can fully understand how to implement the technical solutions disclosed herein according to the above description.

[0221] Those skilled in the art should understand that the embodiments of the present disclosure can be provided as a method, a system, or a computer program product. Therefore, the present disclosure can take the form of an entirely hardware embodiment, an entirely software embodiment, or an embodiment combining software and hardware aspects. Moreover, the present disclosure can take the form of a computer program product implemented on one or more computer-usable non-transitory storage media (including, but not limited to, disk storage, CD-ROMs, optical storage, etc.) containing computer-usable program code.

[0222] The present disclosure is described with reference to flowcharts and / or block diagrams of methods, devices (systems), and computer program products according to embodiments of the present disclosure. It should be understood that the functions specified in one or more flows in the flowcharts and / or one or more blocks in the block diagrams can be implemented by computer program instructions. These computer program instructions can be provided to the processor of a general-purpose computer, a special-purpose computer, an embedded processor, or other programmable data processing apparatus to produce a machine, so that the instructions executed by the processor of the computer or other programmable data processing apparatus produce means for implementing the functions specified in one or more flows in the flowcharts and / or one or more blocks in the block diagrams.

[0223] These computer program instructions can also be stored in a computer readable memory that can direct a computer or other programmable data processing apparatus to function in a particular manner, such that the instructions stored in the computer readable memory produce an article of manufacture including instructions which implement the function specified in the flowchart or flowsheets and / or block or blocks of the block diagrams.

[0224] These computer program instructions can also be loaded onto a computer or other programmable data processing apparatus to cause a series of operational steps to be performed on the computer or other programmable apparatus to produce a computer implemented process such that the instructions which execute on the computer or other programmable apparatus provide steps for implementing the functions specified in the flowchart or flowsheets and / or block or blocks of the block diagrams.

[0225] Although some specific embodiments of the present disclosure have been described in detail above, those skilled in the art should understand that the above examples are only for illustration, and are not intended to limit the scope of the present disclosure. Those skilled in the art should understand that modifications can be made to the above embodiments or equivalent replacements can be made to part of the technical features without departing from the scope and spirit of the present disclosure. The scope of the present disclosure is defined by the appended claims.

Claims

1. A sample analyzer for analyzing a urine sample, comprising: a sample preparation device including a sample carrier portion for carrying a sample container containing a urine sample to be tested of a subject, a sample dispensing portion for aspirating the urine sample to be tested of the subject from the sample carrier portion and discharging it into a reaction container to be assayed, a reagent carrier portion for carrying a detection reagent including a colorimetric reagent and a turbidimetric reagent, a reagent dispensing portion for aspirating the detection reagent from the reagent carrier portion and discharging it into a reaction container to be assayed, and a reaction portion for placing a reaction container so as to incubate an assay solution obtained by reacting the urine sample to be tested with the detection reagent in the reaction container, the assay solution including a first assay solution obtained by reacting the urine sample to be tested with the colorimetric reagent and a second assay solution obtained by reacting the urine sample to be tested with the turbidimetric reagent; an optical detection device including a light detector for detecting a light signal of the assay solution; and a controller configured to: control the sample preparation device to prepare the first assay solution and control the light detector to detect the first assay solution so as to obtain a first urine total protein detection result based on a light signal of the first assay solution; control the sample preparation device to prepare the second assay solution and control the light detector to detect the second assay solution so as to obtain a second urine total protein detection result based on a light signal of the second assay solution; in a case where the urine sample to be tested is proteinuria, calculate a kidney injury marker parameter according to the first urine total protein detection result and the second urine total protein detection result and output the kidney injury marker parameter; and when the kidney injury marker parameter is within a first threshold range, prompt that the kidney of the subject is likely to be in a clinical pathological state of injury. The controller is further configured to differentiate glomerular kidney injury and tubular kidney injury of the subject according to the kidney injury marker parameter. The differentiation of glomerular kidney injury and tubular kidney injury of the subject according to the kidney injury marker parameter includes:

2. The sample analyzer of claim 1, wherein, when the kidney injury marker parameter is within a second threshold range, the subject is differentiated as glomerular kidney injury; and when the kidney injury marker parameter is outside the second threshold range, the subject is differentiated as tubular kidney injury.

3. The sample analyzer of claim 2, wherein, Preferably, when the kidney injury marker parameter is greater than a preset threshold, the subject is differentiated as glomerular kidney injury; and when the kidney injury marker parameter is less than the preset threshold, the subject is differentiated as tubular kidney injury. The controller is further configured to make a risk prediction of tubular kidney injury of the subject according to the kidney injury marker parameter and output a corresponding prediction result. ​ 4. The sample analyzer of any one of claims 1-3, wherein, ​ Preferably, the controller is further configured to predict the subject has a risk of tubular type kidney injury when the kidney injury marker parameter is within a preset threshold range.

5. The sample analyzer of any one of claims 1-4, wherein, The controller is further configured to risk grade the tubular type kidney injury of the subject according to the kidney injury marker parameter, and output the risk grade; preferably, the controller is further configured to risk grade the tubular type kidney injury of the subject into low risk, medium risk and high risk according to the kidney injury marker parameter.

6. The sample analyzer of claim 5, wherein, Risk grading the tubular type kidney injury of the subject into low risk, medium risk and high risk according to the kidney injury marker parameter comprises: outputting a prompt indicating that the tubular type kidney injury of the subject is low risk when the kidney injury marker parameter is within a third threshold range; outputting a prompt indicating that the tubular type kidney injury of the subject is medium risk when the kidney injury marker parameter is within a fourth threshold range and outside the third threshold range; outputting a prompt indicating that the tubular type kidney injury of the subject is high risk when the kidney injury marker parameter is within a fifth threshold range and outside the third threshold range; Preferably, outputting a prompt indicating that the tubular type kidney injury of the subject is low risk when the kidney injury marker parameter is greater than a first threshold; outputting a prompt indicating that the tubular type kidney injury of the subject is medium risk when the kidney injury marker parameter is less than or equal to the first threshold and greater than or equal to a second threshold; and outputting a prompt indicating that the tubular type kidney injury of the subject is high risk when the kidney injury marker parameter is less than the second threshold.

7. The sample analyzer of any one of claims 1-6, wherein, The controller is further configured to: determine whether the urine sample of the subject is proteinuria according to the first total urine protein detection result; or receive input information, and determine whether the urine sample of the subject is proteinuria based on the input information, preferably the input information comprises urine routine information of the subject.

8. The sample analyzer of any one of claims 1-7, wherein, The controller is further configured to prepare the first assay solution and the second assay solution by: controlling the sample preparation device to mix a portion of the urine sample to be tested with the colorimetric reagent to prepare the first assay solution; and controlling the sample preparation device to mix at least a portion of the first assay solution with the turbidimetric reagent to prepare the second assay solution. The controller is further configured to prepare the first assay solution and the second assay solution by:

9. The sample analyzer of any one of claims 1-7, wherein, controlling the sample preparation device to mix a first portion of the urine sample to be tested with the colorimetric reagent to prepare the first assay solution; and controlling the sample preparation device to mix a second portion of the urine sample to be tested with the turbidimetric reagent to prepare the second assay solution. The controller is further configured to: ​ 10. The sample analyzer of any one of claims 1-9, wherein, ​ The renal injury marker parameter is calculated according to a first algorithm, and the first algorithm includes any one of a relative deviation, an absolute deviation, a relative mean deviation, and a weighting of the relative mean deviation of the first urine total protein detection result and the second urine total protein detection result; or The renal injury marker parameter is calculated according to a second algorithm, and the second algorithm includes any one of a relative deviation, an absolute deviation, and a weighting of the relative deviation and the absolute deviation of the logarithm of the first urine total protein detection result and the logarithm of the second urine total protein detection result.

11. The sample analyzer of any of claims 1-10, wherein, The reagent carrying part is configured to place a reagent storage device, and the colorimetric reagent and the turbidimetric reagent are stored in the same reagent storage device.

12. The sample analyzer of any of claims 1-11, wherein, The controller is further configured to control the sample preparation device to prepare the first measurement solution and control the light detector to detect the first measurement solution to obtain the first urine total protein detection result, and control the sample preparation device to prepare the second measurement solution and control the light detector to detect the second measurement solution to obtain the second urine total protein detection result, regardless of whether the to-be-tested urine sample is proteinuria.

13. The sample analyzer of any one of claims 1-11, wherein, The controller is further configured to control the sample preparation device to prepare the first measurement solution and control the light detector to detect the first measurement solution to obtain the first urine total protein detection result, and control the sample preparation device to prepare the second measurement solution and control the light detector to detect the second measurement solution to obtain the second urine total protein detection result, only when it is determined that the to-be-tested urine sample is proteinuria.

14. The sample analyzer of any one of claims 1-11, wherein, The controller is further configured to: First, control the sample preparation device to prepare the first measurement solution and control the light detector to detect the first measurement solution to obtain the first urine total protein detection result; Second, control the sample preparation device to prepare the second measurement solution and control the light detector to detect the second measurement solution to obtain the second urine total protein detection result, only when it is determined that the to-be-tested urine sample is proteinuria, preferably when it is determined that the to-be-tested urine sample is proteinuria according to the first urine total protein detection result.

15. A system for obtaining a renal injury marker parameter, comprising: a first urine detection module configured to perform colorimetric determination and turbidimetric determination on a to-be-tested urine sample of a subject to obtain a first urine total protein detection result based on colorimetric determination and a second urine total protein detection result based on turbidimetric determination; and a data analysis module configured to obtain the first urine total protein detection result and the second urine total protein detection result; when the to-be-tested urine sample is proteinuria, calculate a renal injury marker parameter based on the first urine total protein detection result and the second urine total protein detection result, wherein the renal injury marker parameter reflects the difference between the first urine total protein detection result and the second urine total protein detection result; and output the renal injury marker parameter.

16. The system of claim 15, wherein, The data analysis module is further configured to: when the renal injury marker parameter is within a first threshold range, then the subject is prompted that the subject's kidney can be in a clinical pathologic state of injury; and / or distinguishing glomerular-type renal injury and tubular-type renal injury in the subject according to the renal injury marker parameter; and / or predicting the risk of tubular-type renal injury in the subject according to the renal injury marker parameter, and outputting the corresponding prediction result; and / or risk stratifying tubular-type renal injury in the subject according to the renal injury marker parameter, and outputting the risk stratification.

17. The system of claim 16, wherein, distinguishing glomerular-type renal injury and tubular-type renal injury in the subject according to the renal injury marker parameter, including: when the renal injury marker parameter is within a second threshold range, the subject is identified as having glomerular-type renal injury; and when the renal injury marker parameter is outside the second threshold range, the subject is identified as having tubular-type renal injury; preferably, when the renal injury marker parameter is greater than a preset threshold, the subject is identified as having glomerular-type renal injury; and when the renal injury marker parameter is less than the preset threshold, the subject is identified as having tubular-type renal injury; and / or, predicting the risk of tubular-type renal injury in the subject according to the renal injury marker parameter, and outputting the corresponding prediction result, including: when the renal injury marker parameter is within a preset threshold range, the subject is predicted to have the risk of tubular-type renal injury; and / or, risk stratifying tubular-type renal injury in the subject according to the renal injury marker parameter, and outputting the risk stratification, including: when the renal injury marker parameter is within a third threshold range, a prompt indicating that the subject's tubular-type renal injury is low risk is outputted; when the renal injury marker parameter is within a fourth threshold range and outside the third threshold range, a prompt indicating that the subject's tubular-type renal injury is medium risk is outputted; and when the renal injury marker parameter is within a fifth threshold range and outside the third threshold range and the fourth threshold range, a prompt indicating that the subject's tubular-type renal injury is high risk is outputted; preferably, when the renal injury marker parameter is greater than a first threshold, a prompt indicating that the subject's tubular-type renal injury is low risk is outputted; when the renal injury marker parameter is less than or equal to the first threshold and greater than or equal to a second threshold, a prompt indicating that the subject's tubular-type renal injury is medium risk is outputted; and when the renal injury marker parameter is less than the second threshold, a prompt indicating that the subject's tubular-type renal injury is high risk is outputted.

18. The system of any of claims 15-17, wherein, the data analysis module is further configured to: determine whether the subject's urine sample to be tested is proteinuria according to the first total urine protein detection result.

19. The system of any of claims 15-17, wherein, the data analysis module is further configured to receive input information and determine whether the subject's urine sample is proteinuria based on the input information; Preferably, the system further comprises a second urine testing device for obtaining urine routine information of the subject, and the data analysis module is further configured to obtain the urine routine information and determine whether the urine sample of the subject is proteinuria based on the urine routine information.

20. An analysis method of a urine sample, comprising: obtaining a first urine total protein test result of a subject and a second urine total protein test result of the subject, wherein the first urine total protein test result is obtained by performing a colorimetric method on a to-be-tested urine sample of the subject, and the second urine total protein test result is obtained by performing a turbidimetric method on the to-be-tested urine sample; in the case that the to-be-tested urine sample is proteinuria, calculating a kidney injury marker parameter according to the first urine total protein test result and the second urine total protein test result, wherein the kidney injury marker parameter reflects the difference between the first urine total protein test result and the second urine total protein test result; and outputting the kidney injury marker parameter.

21. The analysis method of a urine sample according to claim 20, further comprising: when the kidney injury marker parameter is within a first threshold range, prompting that the kidney of the subject is likely to be in a clinical pathological state of injury; and / or distinguishing glomerular kidney injury and tubular kidney injury of the subject according to the kidney injury marker parameter; and / or predicting the risk of tubular kidney injury of the subject according to the kidney injury marker parameter, and outputting a corresponding prediction result; and / or risk grading the tubular kidney injury of the subject according to the kidney injury marker parameter, and outputting the risk grading.

22. A computer readable storage medium having stored thereon a computer program, the computer program being executed by a processor to implement the method according to any one of claims 20-21.

23. A sample analyzer for analyzing a urine sample, comprising: a sample preparation device comprising a sample carrying part, a sample dispensing part, a reagent carrying part, a reagent dispensing part, and a reaction part, the sample carrying part is used to carry a sample container, the sample container contains a to-be-tested urine sample of a subject, the sample dispensing part is used to suck the to-be-tested urine sample of the subject from the sample carrying part and discharge it into a to-be-added reaction container, the reagent carrying part is used to carry a detection reagent, the detection reagent comprises a colorimetric reagent and a turbidimetric reagent, the reagent dispensing part is used to suck the detection reagent from the reagent carrying part and discharge it into a to-be-added reagent reaction container, and the reaction part is used to place a reaction container so as to incubate a determination test solution obtained by reaction of the to-be-tested urine sample and the detection reagent in the reaction container, the determination test solution comprises a first determination test solution and a second determination test solution, the first determination test solution is obtained by reaction of the to-be-tested urine sample and the colorimetric reagent, and the second determination test solution is obtained by reaction of the to-be-tested urine sample and the turbidimetric reagent; an optical detection device comprising an optical detector for detecting an optical signal of the determination test solution; and a controller configured to: ​ controlling the sample preparation device to prepare the first assay sample solution, and controlling the light detector to detect the first assay sample solution, so as to obtain a first urine total protein detection result based on a light signal of the first assay sample solution; controlling the sample preparation device to prepare the second assay sample solution, and controlling the light detector to detect the second assay sample solution, so as to obtain a second urine total protein detection result based on a light signal of the second assay sample solution; and outputting the first urine total protein detection result and the second urine total protein detection result.

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