CN116298323B - Biomarker for diagnosing lupus nephritis and application thereof - Google Patents
Biomarker for diagnosing lupus nephritis and application thereof Download PDFInfo
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Abstract
The application relates to a biomarker for diagnosing lupus nephritis and application thereof, belonging to the field of biological medicine. The application provides a biomarker for diagnosing lupus nephritis, which comprises the following steps: an L1-cell adhesion molecule, an extracellular 5 '-nucleotidase, a nucleic acid encoding an L1-cell adhesion molecule, and/or a nucleic acid encoding an extracellular 5' -nucleotidase; the biomarker has strong correlation with lupus nephritis, and diagnosis of the lupus nephritis by using the biomarker has the advantages of high sensitivity and strong specificity.
Description
Technical Field
The application relates to the field of biological medicine, in particular to a biomarker for diagnosing lupus nephritis and application thereof.
Background
Systemic lupus erythematosus (systemic lupus erythematosus, SLE) is a type of multi-system autoimmune disease characterized by persistent inflammation and abnormal formation of autoantibodies, the global overall incidence of systemic lupus erythematosus ranges from 1.5 to 11/10 ten thousand years, and the global prevalence ranges from 13 to 7713.5/10 ten thousand years. One of the severely affected organs in systemic lupus erythematosus is the kidney, lupus Nephritis (LN), which is a major factor in the progression and death of SLE patients, can manifest as abnormalities in the urinary tract, such as hematuria, leukocytosis, cytopenia, and mild proteinuria, or more pronounced manifestations, including nephrotic syndrome or acute nephrotic syndrome or rapid progressive renal failure.
Therefore, timely and accurate diagnosis and treatment is particularly important for improving survival rate of lupus nephritis patients. Diagnosis and therapeutic guidelines for lupus nephritis are typically made by renal biopsy. Due to the invasive techniques used in obtaining tissue samples, complications are numerous, including acute bleeding, infection, and the like. Currently, laboratory markers of lupus nephritis such as proteinuria, creatinine clearance, anti-double stranded DNA autoantibodies (anti-dsDNA) and low plasma complement levels (mainly C3 and C4) have been used to routinely monitor lupus nephritis activity in everyday clinical settings. But these clinical parameters lack sensitivity and specificity reflecting real-time kidney immunopathological activity and chronic kidney tissue damage.
It is clearly necessary to explore novel biomarkers that can distinguish lupus nephritis activity from its severity, predict kidney attacks, monitor therapeutic efficacy and disease progression. Exosomes are microvesicles with the size of about 40-100 nm, are one of the main modes of intercellular communication, and after cells are damaged or become malignant, damaged cells release more exosomes than normal cells, and a large amount of specific information of cells from sources such as proteins, RNA, DNA and the like can be carried in the exosomes; these specific biomarkers can be used as important indicators for disease determination.
The prior art has disclosed that miRNA transmitted by exosomes is used as a diagnosis biomarker of autoimmune diseases, and in addition, exosome-derived proteins can reflect pathological processes related to the diseases, however, most of the markers have the problem of low sensitivity or poor specificity in lupus nephritis diagnosis (see documents: wei Xian, you Yan dance. Research progress of exosomes in lupus nephritis diagnosis [ J ]. The university of medical science of Right river, 2022,44 (06): 914-918.), so new biomarkers with high sensitivity and high specificity are needed to diagnose lupus nephritis at present.
Disclosure of Invention
In order to solve the problems of low sensitivity or poor specificity of the biomarker in the lupus nephritis diagnosis in the prior art, the application provides a biomarker for diagnosing the lupus nephritis, wherein the biomarker comprises an L1-cell adhesion molecule, an extracellular 5 '-nucleotidase, a nucleic acid encoding the L1-cell adhesion molecule and/or a nucleic acid encoding the extracellular 5' -nucleotidase.
Wherein, the NCBI accession number of the L1-cell adhesion molecule (L1 CAM protein, also called CAML1; CD171; HSAS; NCAM-L1; S10; SPG 1) is NP-001265045.1, and the accession number of the nucleic acid encoding the L1-cell adhesion molecule is NG-009645; NCBI accession number for extracellular 5 '-nucleotidase (NT 5E protein, also known as CALJA, CD73, E5 NT) is NP-002517.1 and accession number for nucleic acid encoding extracellular 5' -nucleotidase is NG-028214.
In some embodiments, the biomarker consists of an L1-cell adhesion molecule and an extracellular 5 '-nucleotidase, or the biomarker consists of a nucleic acid encoding an L1-cell adhesion molecule and a nucleic acid encoding an extracellular 5' -nucleotidase.
In some embodiments, the biomarker is derived from an exosome in urine or blood.
In some embodiments, the exosomes comprise exosomes extracted by ultracentrifugation, density gradient centrifugation, ultrafiltration, column chromatography, immunomagnetic bead microsphere methods, or precipitation methods.
The application also provides any one of the following applications of the detection reagent for detecting the level of the biomarker:
a1 Use of a composition for the manufacture of a product for diagnosing lupus nephritis in a subject; or (b)
A2 Use in the preparation of a product for screening a subject for lupus nephritis; or (b)
A3 Use of a composition for the manufacture of a product for assessing the risk of lupus nephritis in a subject; or (b)
A4 Use in the preparation of a product for prognosis evaluation of lupus nephritis.
In some embodiments, the detection reagent comprises a reagent for detecting the expression level of an L1-cell adhesion molecule, a reagent for detecting the expression level of an extracellular 5 '-nucleotidase, a reagent for detecting the expression level of a nucleic acid encoding an L1-cell adhesion molecule, and/or a detection reagent for detecting the expression level of a nucleic acid encoding an extracellular 5' -nucleotidase.
In some embodiments, the detection reagent consists of a reagent for detecting the expression level of an L1-cell adhesion molecule and a reagent for detecting the expression level of an extracellular 5 '-nucleotidase, or the detection reagent consists of a reagent for detecting the expression level of a nucleic acid encoding an L1-cell adhesion molecule and a detection reagent for detecting the expression level of a nucleic acid encoding an extracellular 5' -nucleotidase.
In some embodiments, the detection reagent comprises a detection reagent that detects the biomarker by flow cytometry, western blot, immunofluorescence, enzyme-linked immunosorbent assay, mass spectrometry, near infrared spectrometry, immunochromatography, capillary gel electrophoresis, or colloidal gold immunoassay.
In some embodiments, the detection reagent comprises an antibody that binds to an L1-cell adhesion molecule, an antibody that binds to an extracellular 5 '-nucleotidase, a primer for specifically amplifying a nucleic acid encoding an L1-cell adhesion molecule, and/or a primer for specifically amplifying a nucleic acid encoding an extracellular 5' -nucleotidase.
The application also provides a kit comprising a detection reagent for detecting the biomarker, wherein the kit has at least one of the following purposes:
b1 Use of a composition for the manufacture of a product for diagnosing lupus nephritis in a subject; or (b)
B2 Use in the preparation of a product for screening a subject for lupus nephritis; or (b)
B3 Use of a composition for the manufacture of a product for assessing the risk of lupus nephritis in a subject; or (b)
B4 Use in the preparation of a product for prognosis evaluation of lupus nephritis.
In some embodiments, the detection reagent comprises a reagent for detecting the expression level of an L1-cell adhesion molecule, a reagent for detecting the expression level of an extracellular 5 '-nucleotidase, a reagent for detecting the expression level of a nucleic acid encoding an L1-cell adhesion molecule, and/or a detection reagent for detecting the expression level of a nucleic acid encoding an extracellular 5' -nucleotidase.
In some embodiments, the detection reagent consists of a reagent for detecting the expression level of an L1-cell adhesion molecule and a reagent for detecting the expression level of an extracellular 5 '-nucleotidase, or the detection reagent consists of a reagent for detecting the expression level of a nucleic acid encoding an L1-cell adhesion molecule and a detection reagent for detecting the expression level of a nucleic acid encoding an extracellular 5' -nucleotidase.
In some embodiments, the detection reagent comprises a detection reagent that detects the biomarker by flow cytometry, western blot, immunofluorescence, enzyme-linked immunosorbent assay, mass spectrometry, near infrared spectrometry, immunochromatography, capillary gel electrophoresis, or colloidal gold immunoassay.
In some embodiments, the detection reagent comprises an antibody that binds to an L1-cell adhesion molecule, an antibody that binds to an extracellular 5 '-nucleotidase, a primer for specifically amplifying a nucleic acid encoding an L1-cell adhesion molecule, and/or a primer for specifically amplifying a nucleic acid encoding an extracellular 5' -nucleotidase.
In some embodiments, the test object of the test agent is urine or blood. When the kit is used, urine or blood is collected for exosome extraction, and then the level of the biomarker on the extracted exosome is detected. The extraction method of exosomes comprises the following steps: ultracentrifugation, density gradient centrifugation, ultrafiltration, column chromatography, immunomagnetic bead microsphere or precipitation.
The application also provides application of the biomarker serving as a target in preparing a medicament for treating or preventing lupus nephritis.
The application has the beneficial effects that:
1. the present application provides a biomarker for diagnosing lupus nephritis, comprising an L1-cell adhesion molecule from an exosome, an extracellular 5 '-nucleotidase, a nucleic acid encoding the L1-cell adhesion molecule, and/or a nucleic acid encoding the extracellular 5' -nucleotidase. The research shows that the expression level of the L1-cell adhesion molecule and the extracellular 5' -nucleotidase in urine exosomes of lupus nephritis patients is obviously higher than that of normal people. The L1-cell adhesion molecule and/or the extracellular 5 '-nucleotidase have strong correlation with lupus nephritis, so that the diagnosis of the lupus nephritis is carried out by taking the L1-cell adhesion molecule and/or the extracellular 5' -nucleotidase as detection objects, and the sensitivity of independently detecting the L1-cell adhesion molecule is 80 percent, and the specificity is 73.33 percent; the sensitivity of detecting the extracellular 5' -nucleotidase alone is 76.67% and the specificity is 80%; the sensitivity of the detection of the L1-cell adhesion molecule and the extracellular 5' -nucleotidase which are simultaneously used as biomarkers is 90 percent, and the specificity is 93.33 percent; in addition, the area under the curve of the subjects with lupus nephritis is identified by using the L1-cell adhesion molecules alone (AUC, which is used for measuring the performance of the classifier, the AUC value is closer to 1, which indicates that the performance of the classifier is better, whereas the AUC value is closer to 0, which indicates that the performance of the classifier is poorer.) is 0.832, the AUC of lupus nephritis is identified by using the extracellular 5' -nucleotidase alone is 0.868, the AUC of lupus nephritis is identified by using the combination of the L1-cell adhesion molecules and the extracellular 5' -nucleotidase, the AUC of the three results is better, and the AUC effect of the combination of the L1-cell adhesion molecules and the extracellular 5' -nucleotidase is optimal, so that the biomarker provided by the application has good application prospect in the aspect of diagnosing the lupus nephritis.
2. The application provides a kit for diagnosing lupus nephritis, which comprises a detection reagent for detecting the biomarker, and has at least one of the following purposes:
b1 Use of a composition for the manufacture of a product for diagnosing lupus nephritis in a subject; or (b)
B2 Use in the preparation of a product for screening a subject for lupus nephritis; or (b)
B3 Use of a composition for the manufacture of a product for assessing the risk of lupus nephritis in a subject; or (b)
B4 Use in the preparation of a product for prognosis evaluation of lupus nephritis;
the detection object of the kit has strong correlation with lupus nephritis, so the diagnosis of the lupus nephritis is carried out by using the kit, the sensitivity of the kit for independently detecting the L1-cell adhesion molecule is 80 percent, and the specificity is 73.33 percent; the sensitivity of detecting the extracellular 5' -nucleotidase alone is 76.67% and the specificity is 80%; the sensitivity of the detection of the L1-cell adhesion molecule and the extracellular 5' -nucleotidase which are simultaneously used as biomarkers is 90 percent, and the specificity is 93.33 percent; in addition, the AUC of the lupus nephritis is identified by using the L1-cell adhesion molecule alone and is 0.832, the AUC of the lupus nephritis is identified by using the extracellular 5' -nucleotidase alone and is 0.868, the AUC of the lupus nephritis is identified by using the L1-cell adhesion molecule and the extracellular 5' -nucleotidase in combination and is 0.947, the AUC of the three results are good, and the AUC effect of the lupus nephritis is identified by using the L1-cell adhesion molecule and the extracellular 5' -nucleotidase in combination, so that the kit provided by the application has good application prospect in the aspect of diagnosing the lupus nephritis.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present application, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a scatter plot of mass spectra of healthy and lupus nephritis patient groups, wherein-Log 10P values represent Log values of P, log2 (lupus nephritis/healthy) represents Log values of fold change in protein fold change, up-right-most is protein expressing significantly up-regulated protein, up-left is protein expressing significantly down-regulated protein, and middle part is protein with insignificant and relatively stable expression difference.
FIG. 2 is a graph showing the comparison of the expression levels of L1CAM and NT5E, L CAM & NT5E in the lupus nephritis patient group and the healthy group, wherein L1CAM represents the result of L1-cell adhesion molecule detection of a sample, NT5E represents the result of extracellular 5 '-nucleotidase detection of a sample, L1CAM & NT5E represents the result of simultaneous L1-cell adhesion molecule and extracellular 5' -nucleotidase detection of a sample, the abscissa represents the type of subject, the healthy group and the lupus nephritis patient group are classified, and the ordinate represents the number of exosomes positive for the relevant biomarker in every 1 ten thousand exosomes.
FIG. 3 is an analysis of ROC curves of subjects for L1CAM, NT5E, L1CAM & NT5E index detection, plotted on the abscissa with 100% -specificity as a parameter and on the ordinate with sensitivity as a parameter.
Detailed Description
Experimental example 1: acquisition of molecular markers
1. Mass spectrometry detection
Collecting urine of healthy people and lupus nephritis patients, extracting exosomes (a sample is taken from Nanjing drummer Hospital), extracting proteins, quantifying, and performing mass spectrometry detection; among them, protein extraction and mass spectrometry detection are described in the literature: HOSHINO, AYUKO, KIM, HAN SANG, BOJMAR, LINDA, et al Extracellular Vesicle and Particle Biomarkers Define Multiple Human Cancers [ J ]. Cell,2020,182 (4): 1044- +. DOI 10.1016/J. Cell.2020.07.009.
FIG. 1 shows the results of mass spectrometry detection, wherein the L1CAM protein and the NT5E protein were selected from the lupus nephritis group up-regulation proteins.
2. Mass spectrometry data analysis
The mass spectrum results are subjected to qualitative and quantitative analysis of protein by a quality control machine, a filtering machine and a data normalization machine, the results are shown in a graph of comparing the expression quantity of L1CAM, NT5E, L1CAM & NT5E in lupus nephritis patient groups and healthy group pairing groups, wherein the abscissa is the type of a subject, the subjects are divided into healthy person groups and lupus nephritis patient groups, the ordinate is the number of the exosomes positive to the relevant biomarker in every 1 ten thousand exosomes, the L1CAM and NT5E proteins have significant differences between the lupus nephritis patient groups and the healthy groups, and the L1CAM & NT5E differences are particularly significant.
Experimental example 2: verification of molecular markers
1. Sample collection
Urine samples of healthy people who meet the standard and live lupus nephritis patients are collected by a Standard Operation Procedure (SOP), and complete clinical case information data are collected by the system. The collected samples were 30 healthy human samples (non-lupus patients, non-lupus nephritis patients), 30 lupus nephritis patient samples (lupus nephritis diagnosed by clinical gold standard), and all the collected healthy human samples were matched with the patients in terms of age and sex.
2. Extraction and purification of exosomes
Exosome extraction was performed using commercially available exosome extraction reagents (available from thermo, cat# 4484452) as follows:
(1) firstly, a sample (fresh or frozen sample, wherein the frozen sample needs to be thawed at room temperature of 25 ℃) is centrifuged for 10min at 4 ℃ and 3000 Xg;
(2) transferring the supernatant to a new centrifuge tube, centrifuging at 4 ℃ and 12000 Xg for 15 min, discarding the precipitate, and transferring the supernatant to the new centrifuge tube;
(3) placing the pretreated sample on ice, adding 1/4 volume of exosome extraction reagent, fully and uniformly mixing, and standing on ice for 20 minutes;
(4) after standing, centrifuging the sample at 4 ℃ for 60 min at 10,000Xg, discarding the supernatant, and collecting the precipitate rich in exosomes;
(5) the pellet was resuspended in PBS, centrifuged at 12000 Xg for 2 min at 4℃and the pellet was discarded and the supernatant, the exosome-rich solution, was retained.
3. Flow cytometry detection
(1) The supernatant obtained above was dissolved in an equal volume of PBS solution containing 2% (volume ratio 2:98) BSA, then 2. Mu.L of primary anti-L1CAM (anti-L1 CAM, available from abcam, cat. No. ab 270455) and primary anti-NT5E (anti-NT 5E, available from abcam, cat. No. ab 202122) were added respectively, mixed, incubated at room temperature (25 ℃) for 30 minutes, then a goat anti-mouse secondary anti-PE (goat anti-mouse, available from abcam, cat. No. ab 97024) was added to the L1CAM, and 1. Mu.L of FITC-labeled goat anti-rabbit secondary anti-rabbit (goat anti-rabit, available from abcam, cat. No. ab 102344) was added to the NT5E, and after mixing, incubated at room temperature (25 ℃) for 30 minutes in the dark, the incubated sample was obtained. All antibodies and reagents were titrated and diluted to prevent autofluorescence.
(2) Parameter correction for flow cytometry
A CytoFLEX S model flow cytometer from Beckman Coulter company, usa was used. Firstly, adjusting parameters of a flow cytometer to detect particles with the size of exosomes, calibrating the sizes of the particles by using microspheres, and specifically, establishing an FSC/VSSC scatter diagram; microspheres with the sizes of 100nm, 200nm, 300nm, 400nm and 500nm are used for starting the machine, and the door is set. Under the same condition, detecting the proportion or quantity of particles of the sample to be detected under the corresponding fluorescence, and quantifying the expression quantity of the positive exosome in the fixed quantity.
(3) Flow cytometry detection
And (3) detecting the incubated sample on a machine, and analyzing the detection result by using flow cytometry analysis software.
Referring to table 1, the detection results of different detection indexes are true positive if the indexes are positive and the diseases are negative, and the true negative if the indexes are negative and the diseases are negative; if the index negative corresponds to the disease positive, the result is false negative, and similarly if the index positive corresponds to the disease negative, the result is false positive.
TABLE 1 detection results of different detection indicators
The calculation method of the detection sensitivity and specificity is as follows:
sensitivity = true positive number/(true positive number + false negative number) ×100%.
Specificity = true negative population/(true negative population + false positive population) ×100%. Wherein, the true positive, false negative, true negative and false positive are all detection values.
Sensitivity of L1CAM protein: 24/(24+6) =80%;
specificity of the L1CAM protein: 22/(24+6) =73.33%;
sensitivity of NT5E protein: 23/(24+6) =76.67%;
specificity of NT5E protein: 24/(24+6) =80%;
sensitivity of L1CAM & NT5E protein: 27/(24+6) =90%;
specificity of L1CAM & NT5E protein: 28/(24+6) =93.33%.
Referring to Table 2 for comparison of the sensitivity and specificity of the different detection markers, it can be seen from Table 2 that the sensitivity of L1CAM protein alone was 80% and the specificity was 73.33%; the sensitivity of the single detection of NT5E protein was 76.67% and the specificity was 80%; the sensitivity of the combined detection of the L1CAM protein and the NT5E protein was 90% and the specificity was 93.33%. Therefore, the L1CAM or the NT5E can be detected independently to distinguish healthy people from lupus nephritis patients to a certain extent, but the combined detection effect of the healthy people and the lupus nephritis patients is better, which shows that the combined detection analysis and diagnosis effect of the L1CAM and the NT5E is obvious.
TABLE 2 sensitivity and specificity comparison of different detection indicators
The L1CAM & NT5E joint detection analysis shows that the biomarker from exosomes and/or the reagent for detecting the biomarker provided by the application has better sensitivity and specificity in the diagnosis aspect of lupus nephritis patients, and has good clinical application prospect.
In addition, in the graph of the working characteristic curve (Receiver Operating Characteristic Curve, ROC curve, see fig. 3) of the analysis subject, area refers to the Area under AUC (Area Under ROC Curve) ROC curve, AUC of the individual identification of the L1CAM protein in the urine exosome sample for lupus nephritis is 0.832, AUC of the individual identification of the NT5E protein in the urine exosome sample for lupus nephritis is 0.868, AUC of the individual identification of the L1CAM and NT5E proteins in the urine exosome sample for lupus nephritis is 0.947, AUC of the three results are good, and AUC effect of the individual identification of the L1CAM and NT5E proteins for lupus nephritis is optimal.
Similarly, based on the biomarker combination, the further developed detection reagent and detection kit have more accurate prompt effect on lupus nephritis with slightly difficult detection of clinical symptoms, and have extremely high medical and practical values.
Example 1: biomarker for diagnosing lupus nephritis
The present embodiment provides a biomarker for diagnosing lupus nephritis, the biomarker comprising: l1-cell adhesion molecules, extracellular 5 '-nucleotidase, nucleic acids encoding L1-cell adhesion molecules and/or nucleic acids encoding extracellular 5' -nucleotidase.
Example 2: kit for diagnosing lupus nephritis
The embodiment provides a kit for diagnosing lupus nephritis, which comprises a detection reagent for detecting the biomarker, and has at least one of the following purposes:
b1 Use of a composition for the manufacture of a product for diagnosing lupus nephritis in a subject; or (b)
B2 Use in the preparation of a product for screening a subject for lupus nephritis; or (b)
B3 Use of a composition for the manufacture of a product for assessing the risk of lupus nephritis in a subject; or (b)
B4 Use in the preparation of a product for prognosis evaluation of lupus nephritis.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the application.
Claims (7)
1. Use of any one of the following detection reagents for detecting the level of a biomarker:
a1 Use of a composition for the manufacture of a product for diagnosing lupus nephritis in a subject; or (b)
A2 Use in the preparation of a product for screening a subject for lupus nephritis; or (b)
A3 Use of a composition for the manufacture of a product for assessing the risk of lupus nephritis in a subject; or (b)
A4 Use in the preparation of a product for prognosis evaluation of lupus nephritis;
the biomarker is: an L1-cell adhesion molecule and/or a nucleic acid encoding an L1-cell adhesion molecule; or alternatively
The biomarker is: l1-cell adhesion molecules and/or nucleic acids encoding L1-cell adhesion molecules, and extracellular 5 '-nucleotidase and/or nucleic acids encoding extracellular 5' -nucleotidase;
the biomarker is derived from exosomes in urine.
2. The use according to claim 1, wherein the detection reagent comprises a reagent for detecting the expression level of an L1-cell adhesion molecule and/or a nucleic acid encoding an L1-cell adhesion molecule; or alternatively
Detection reagents for detecting the expression level of the L1-cell adhesion molecule and/or nucleic acid encoding the L1-cell adhesion molecule, and the extracellular 5 '-nucleotidase and/or nucleic acid encoding the extracellular 5' -nucleotidase.
3. The use according to claim 1, wherein the detection reagent comprises a detection reagent for detecting the level of the biomarker by flow cytometry, western immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay, mass spectrometry, near infrared spectroscopy, immunochemistry luminescence, capillary gel electrophoresis or colloidal gold immunoassay.
4. The use according to claim 1, wherein the detection reagent comprises an antibody that binds to an L1-cell adhesion molecule and/or a primer for specifically amplifying a nucleic acid encoding an L1-cell adhesion molecule; or alternatively
The detection reagent comprises: an antibody that binds to an L1-cell adhesion molecule and/or a primer for specifically amplifying a nucleic acid encoding an L1-cell adhesion molecule, and an antibody that binds to an extracellular 5 '-nucleotidase and/or a primer for specifically amplifying a nucleic acid encoding an extracellular 5' -nucleotidase.
5. Use of a detection reagent for detecting a biomarker in the preparation of a kit, the use comprising:
b1 Use of a composition for the manufacture of a product for diagnosing lupus nephritis in a subject; or (b)
B2 Use in the preparation of a product for screening a subject for lupus nephritis; or (b)
B3 Use of a composition for the manufacture of a product for assessing the risk of lupus nephritis in a subject; or (b)
B4 Use in the preparation of a product for prognosis evaluation of lupus nephritis;
the biomarker is: an L1-cell adhesion molecule and/or a nucleic acid encoding an L1-cell adhesion molecule; or alternatively
The biomarker is: l1-cell adhesion molecules and/or nucleic acids encoding L1-cell adhesion molecules, and extracellular 5 '-nucleotidase and/or nucleic acids encoding extracellular 5' -nucleotidase;
the biomarker is derived from exosomes in urine.
6. The use according to claim 5, wherein the detection reagent comprises a reagent for detecting the expression level of an L1-cell adhesion molecule and/or a nucleic acid encoding an L1-cell adhesion molecule;
or the detection reagent comprises: detection reagents for detecting the expression level of an L1-cell adhesion molecule and/or a nucleic acid encoding an L1-cell adhesion molecule, and a 5 '-nucleotidase and/or a nucleic acid encoding an extracellular 5' -nucleotidase.
Application of L1-cell adhesion molecules and/or nucleic acid encoding the L1-cell adhesion molecules as targets in preparing medicines for treating or preventing lupus nephritis.
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