Nucleic acid aptamer targeting klm-1 and application thereof

Abstract

The application discloses a nucleic acid aptamer for targeting KIM-1 and application thereof. The nucleic acid aptamer for targeting KIM-1 is at least one of a nucleic acid aptamer K3 with a nucleic acid sequence as shown in SEQ ID NO. 1 and a nucleic acid aptamer K34 with a nucleic acid sequence as shown in SEQ ID NO. 2. Compared with a KIM-1 monoclonal antibody, the nucleic acid aptamer has high specificity and high affinity, and has the advantages of weak immunogenicity, small side effect, large-batch artificial synthesis, short cycle, low cost, strictly controllable quality and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to a nucleic acid aptamer targeting KIM-1 and application thereof. BACKGROUND

[0002] However, it is difficult for clinical medical workers to diagnose, intervene and treat early kidney disease in actual medical work, mainly because the clinical symptoms of patients with early kidney damage are not significant, leading to misdiagnosis and missed diagnosis, and it is easy to delay the disease (Xiao Z, Huang Q, Yang Y, et al. Emerging early diagnostic methods for acute kidney injury. Theranostics. 2022; 12 (6), 2963-2986.).

[0003] Several indicators such as urine routine, osmotic pressure, blood creatinine, urea nitrogen and endogenous creatinine clearance rate are used to judge kidney damage clinically, but in most cases, when these indicators increase, many kidney damages are already very serious or have produced irreversible damage (Oh DJ. A long journey for acute kidney injury biomarkers. Ren Fail. 2020 Nov; 42 (1), 154-165.). The best method to treat kidney damage is prevention. Early kidney damage is reversible, and timely discovery, early intervention and treatment are beneficial to prevent the occurrence and control the progression of the disease. Therefore, a large number of scientists continue to study methods or biomarkers that can be used for early kidney damage diagnosis.

[0004] KIM-1 (also known as HAVCR1 or TIM-1) is a glycosylated transmembrane protein, which is not expressed or lowly expressed in normal kidney, but highly expressed in proximal tubular cells when kidney injury occurs (Karmakova capital TeCAC, Sergeeva NS, Kanukoev capital Ka C, Alekseev BY Kaprin capital A C. Kidney Injury Molecule 1 (KIM-1): a Multifunctional Glycoprotein and Biological Marker (Review). Sovrem Tekhnologii Med. 2021; 13(3), 64-78.). The expression of KIM-1 in proximal tubular cells is significantly increased in kidney tissue biopsy of patients with acute kidney injury (Yin W, Kumar T, Lai Z, et al. Kidney injury molecule-1, asensitive and specific marker for identifying acute proximal tubular injury, can be used to predict renal functional recovery in native renal biopsies. Int Urol Nephrol. 2019 Dec; 51(12), 2255-2265.). Koyner JL et al. studied the expression of KIM-1 in various kidney diseases in humans, and analyzed whether the concentration of KIM-1 in urine was correlated with the expression of KIM-1 in kidney, kidney tissue injury, kidney function and proteinuria. The results showed that the expression of KIM-1 was increased in all kidney diseases except for minimal change nephropathy, which was positively correlated with serum creatinine and negatively correlated with creatinine clearance rate, and had no correlation with urinary protein (Koyner JL, Vaidya VS, Bennett MR, et al. Urinary biomarkers in the clinical prognosis and early detection of acute kidney injury. Clin J Am Soc Nephrol. 2010 Dec; 5(12), 2154-65.).KIM-1 is extremely stable in urine, and the level of urine KIM-1 is consistent with the degree of pathological injury, which indicates that urine KIM-1 has high value for early diagnosis of kidney injury (Vaidya VS, Ford GM, Waikar SS, et al. A rapid urine test for early detection of kidney injury. Kidney Int. 2009 Jul; 76 (1), 108-14.).

[0005] In view of the clinical detection needs of KIM-1, different commercial companies have developed monoclonal antibody-based detection kits for detecting KIM-1 protein in serum and urine samples, but we have purchased almost all KIM-1 detection kits on the market, and found that most of them are only used for scientific research, and have the problems of low specificity and sensitivity. We also tried to prepare monoclonal antibodies, and found that the effect was general, mainly because KIM-1 protein is a membrane protein, and is a highly glycosylated protein, resulting in a weak immunogen type, and no matter how to increase the amount or change the type of immunoadjuvant, the effect is not good. SUMMARY

[0006] The primary purpose of the present application is to overcome the shortcomings and deficiencies of the prior art, and to provide a nucleic acid aptamer targeted to bind KIM-1.

[0007] Another purpose of the present application is to provide the use of the above-mentioned nucleic acid aptamer targeted to bind KIM-1.

[0008] The purpose of the present application is achieved by the following technical solutions:

[0009] A nucleic acid aptamer targeted to bind KIM-1 is at least one of a nucleic acid aptamer K3 with a nucleic acid sequence as shown in SEQ ID NO. 1 and a nucleic acid aptamer K34 with a nucleic acid sequence as shown in SEQ ID NO. 2.

[0010] K3: 5'-CACGTCCGTCCCCCTCACCGCTGCGTCCCCCTCATC-3';

[0011] K34: 5'-CCCCTCCCGCCGATCCCTCGCTCTTTAGCCTGACCT-3'.

[0012] The above-mentioned nucleic acid aptamer targeted to bind KIM-1 is a labeled nucleic acid aptamer.

[0013] The labeled substance includes colloidal gold, biotin and fluorescent molecules.

[0014] The fluorescent molecules are preferably at least one of FAM and Cy3.

[0015] The nucleic acid aptamer targeting KIM-1 is used for preparing a KIM-1 detection reagent.

[0016] The present application has the following advantages and effects compared with the prior art:

[0017] The DNA nucleic acid aptamer provided by the present application for recognizing and binding KIM-1 protein has high specificity and high affinity compared with the monoclonal antibody developed by us in the early stage, and the DNA nucleic acid aptamer has weak immunogenicity and small side effects. The nucleic acid aptamer can be artificially synthesized in large quantities, has short cycle and low cost, and the quality is strictly controllable. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 is a target cell identification result graph.

[0019] Figure 2 is a nucleic acid aptamer screening efficiency identification result graph.

[0020] Figure 3 is a flow cytometry screening result graph of the nucleic acid aptamer capable of binding to the target cell.

[0021] Figure 4 is a flow cytometry detection result graph of the binding of different concentrations of FAM-labeled nucleic acid aptamer to the target cell.

[0022] Figure 5 is a nucleic acid aptamer Kd value detection result graph.

[0023] Figure 6 is a Cy3-labeled nucleic acid aptamer labeled target cell and control cell fluorescence photo graph. DETAILED DESCRIPTION

[0024] The present application will be further described in detail below in combination with examples and drawings, but the embodiments of the present application are not limited thereto.

[0025] Example 1

[0026] The screening is performed by using the CELL-SELEX technology, and the steps include: in the first step, the nucleic acid aptamer library is incubated with reverse screening cells not expressing or lowly expressing KIM-1, and the unbound nucleic acid aptamer is reserved; in the second step, the nucleic acid aptamer reserved in the first step is incubated with target cells highly expressing KIM-1, the unbound oligonucleotide is removed, and the nucleotide bound on the target cells is collected; in the third step, the nucleic acid aptamer capable of binding the target cells obtained in the second step is subjected to PCR amplification, single-stranded is prepared by asymmetric PCR, and the library available for re-screening is obtained; the above is one screening cycle, through the above cycle steps, several rounds of screening are performed, so that the nucleic acid aptamer with high affinity is enriched, finally the nucleic acid is sequenced, ranked according to the enrichment abundance, then the top 70 ranked ones are selected for verification and identification, and finally the nucleic acid aptamer targeting KIM-1 is obtained. The specific screening steps used in the present application are as follows:

[0027] I. Library information:

[0028] All the nucleic acid sequences (such as libraries, primers, etc.) involved in the present application are synthesized by the Shengong Bioengineering (Shanghai) Co., Ltd. as shown in Table 1.

[0029] Table 1

[0030]

[0031] II. Construction of target cells:

[0032] Human renal tubular epithelial cells HK-2 (purchased from Shanghai Fuheng Biotechnology Co., Ltd.) are inoculated in F12 culture medium containing 10% v / v fetal bovine serum, cultured in vitro for three generations, and after stabilization, 10, 20, 30, 40 and 50 μg / mL vancomycin-containing F12 culture solution is added respectively for further culture for 24 h. Part of the cells are taken to detect the expression of KIM-1 protein by Western Blot and the expression of KIM-1 mRNA by qPCR.

[0033] The results are shown in Figure 1 It can be seen that the expression amount of KIM-1 is positively correlated with the action concentration of vancomycin. Since vancomycin causes certain damage to cells and affects the growth of cells, the HK-2 cells treated by 30 μg / mL vancomycin are selected for clonal growth to obtain target cells.

[0034] III. Related experimental system

[0035] The composition of the QPCR mixture solution is shown as follows: ddH2O 866 μL, 10x enzyme buffer 100 μL, dNTP mixture (10 mM) 20 μL, Lib1 S1 (100 μM) 5 μL, Lib2 A2 (100 μM) 5 μL, Taq Plus enzyme 4 μL (50 U), EvaGreen 40 μL.

[0036] The composition of the PCR mixture solution for preparing single-stranded DNA (ssDNA) is shown as follows: ddH2O 866 μL, 10x enzyme buffer 100 μL, dNTP mixture (10 mM) 20 μL, Lib1 S1-FAM (100 μM) 5 μL, Lib2 A2-polyA (100 μM) 5 μL, Taq Plus enzyme 4 μL (50 U).

[0037] Four, specific screening process

[0038] 1. Resuscitation of HK-2 and target cells prepared in step two (i.e. HK-2 cells incubated with 30 μg / mL vancomycin for 24 h):

[0039] (1) Prepare F12 complete medium: inactivate fetal bovine serum (PAN, Argentina) at 65°C for 30 minutes, and then add the inactivated fetal bovine serum to F12 medium in an ultraclean workbench to a final concentration of 10% v / v. The prepared medium is stored in a refrigerator at 4°C for standby;

[0040] (2) Quickly take the frozen HK-2 and target cells from the -80°C refrigerator, quickly put them into a 37°C water bath to melt the cells, and carefully shake the frozen tube to accelerate cell lysis; in an ultraclean workbench, transfer the melted cells to a 15 mL centrifuge tube pre-added with 2 mL F12 complete medium, and carefully blow and mix;

[0041] (3) After centrifugation at 1000 rpm in a centrifuge at room temperature for 5 min, discard the supernatant in an ultraclean workbench, resuspend the cells with 1 mL F12 complete medium, and then transfer them to a 60 mm culture dish and supplement the medium to 4 mL;

[0042] (4) Label the cell name, date, operator and cell generation number on the medium;

[0043] (5) Observe the cells under an inverted microscope, and transfer the culture dish containing the cells to a 37°C, 5% CO2 incubator for culture.

[0044] 2. Cell passage:

[0045] (1) Take out the target cells and counter-screening cells (untreated HK-2 cells) from the constant temperature incubator, and observe the state of the cells under an inverted microscope. The cells are normal.

[0046] (2) Transfer the cells to a sterile 15 mL centrifuge tube in a super-clean bench, and centrifuge at 1000 rpm for 5 min at room temperature.

[0047] (3) Transfer the centrifuge tube to the super-clean bench, discard the cell supernatant, resuspend the cells with 1 mL of fresh complete medium, then transfer the cells to a culture dish, and add fresh medium to 4 mL.

[0048] (4) Label the cell name, date, operator, and cell passage number on the culture dish.

[0049] (5) Observe the cells under an inverted microscope, and transfer the culture dish containing the cells to a 37°C, 5% CO2 constant temperature incubator for further culture. Observe the cells daily, and passage the cells regularly. From the start of the experiment with P3 target cells (30 μg / mL vancomycin treatment for 24 h) and counter-screening cells (PBS control), each passage is used in turn for the next round of screening. After trypsin digestion of the target cells and counter-screening cells, count the cells on a cell counter, and uniformly count to 3 × 10 5 / mL, then start the screening experiment.

[0050] 3. Screening process: first round of screening

[0051] (1) Take one branch of the synthesized lib2-76nt library dry powder, centrifuge at 14000 g for 10 min, add 137 μL DPBS, vortex to dissolve the dry powder, then centrifuge at 14000 g for 10 min, and distribute to PCR tubes.

[0052] (2) Place in a PCR instrument for denaturation and renaturation. The program is: 95°C for 10 min, immediately ice water bath for 5 min, and balance to room temperature.

[0053] (3) Add the denatured and renatured library to the counter-screening cells, mix gently with a pipette gun, and incubate on ice for 60 min.

[0054] (4) After centrifugation at 1000 rpm for 5 min at room temperature, aspirate the supernatant with a gun head and mark it as pool-;

[0055] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is marked as wash1-;

[0056] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is marked as wash2-;

[0057] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is recorded as wash3-;

[0058] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is recorded as wash4-;

[0059] Add 200 μL ultrapure water to the cells, boil in a water bath for 10 min, centrifuge at 10000 rpm for 2 min at room temperature, and the supernatant is recorded as Elution-.

[0060] (5) Add pool- to the target cells, mix slowly with a pipette gun, and incubate on ice for 60 min on a shaker;

[0061] After centrifugation at 1000 rpm for 5 min at room temperature, aspirate the supernatant with a gun head and record it as pool+.

[0062] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is recorded as wash1+;

[0063] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is recorded as wash2+;

[0064] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is recorded as wash3+;

[0065] Rinse the cells with 200 μL DPBS, centrifuge at 1000 rpm for 5 min at room temperature, and the supernatant is recorded as wash4+;

[0066] Add 200 μL ultrapure water to the cells, boil in a water bath for 10 min, centrifuge at 10000 rpm for 2 min at room temperature, and the supernatant is recorded as Elution+.

[0067] (6) Take Roche 8-tube PCR tubes, add 30 μL QPCR mixed solution to each well, and then add 2 μL of Elution- and Elution+ respectively, and perform fluorescent quantitative PCR according to the following procedure: 95°C for 2 min; 95°C for 0.5 min, 60°C for 0.5 min, 72°C for 0.5 min, 25 cycles.

[0068] 4. Preparation of single-stranded DNA:

[0069] (1) Take 2 mL of PCR mixed solution for preparing single-stranded DNA from -20°C, add it to the remaining Elution+, and mix well in a 50 mL centrifuge tube. Add 8 mL of EM90 oil and place it on a high-power vortex shaker to prepare an emulsion.

[0070] (2) Aliquot the emulsion into PCR tubes, 90 μL per tube, and perform PCR for 25 cycles. The program is: 95℃ for 2 min; 95℃ for 1 min, 60℃ for 1 min, 72℃ for 1 min, for 25 cycles.

[0071] (3) Recover PCR products.

[0072] (4) Concentrate the PCR product with n-butanol. Transfer the PCR product to a 10 mL centrifuge tube, fill it with n-butanol, mix well, and centrifuge at 10000 g for 10 min. After centrifugation, separate the layers, remove the clear upper layer, and transfer the lower layer of amplification product to a small EP tube (approximately 100 μL). Take 90 μL and transfer it to the small centrifuge tube, add 100 μL of urea loading buffer, mix well, and heat the PCR instrument at 95 °C for 10 min.

[0073] (5) Denaturing PAGE electrophoresis was used to separate single strands, the gel was cut, boiled, and ssDNA was concentrated with n-butanol; then ssDNA was dialyzed overnight with DPBS in a 3.5KD dialysis bag and the concentration was determined by micro-UV.

[0074] In rounds 2, 3, 4, 5 and 6, the selection criteria change in each round, as shown in Table 2, while the other steps remain the same.

[0075] Table 2

[0076]

[0077] V. Document retention rate:

[0078] The retention rates of the six-round library are shown in Table 3 below:

[0079] Table 3

[0080]

[0081] Analysis of retention rates in each round of screening, as follows: Figure 2 As shown, from Figure 2 It can be seen that each round of screening can effectively obtain the corresponding nucleic acid aptamers. As the number of rounds increases, the nucleic acid aptamers that bind to the target cells gradually become richer, while the nucleic acid aptamers that do not bind or have low binding power gradually decrease.

[0082] VI. Flow cytometry FC-MS detection of library affinity:

[0083] 1. Sample preparation: P10 generation target cells were digested with trypsin and divided into 6 groups. Cells were counted using a cell counter, with each group counted at a uniform rate of 3 × 10⁻⁶. 5 / mL, control 1 (PBS) and 50 μL of control 2 (pool 0), pool 1, pool 4, pool 5 and pool 6 (each at a concentration of 400 nM) were added to the HK-2 cells, mixed, and incubated on ice for 40 min;

[0084] 2. Turn on the flow cytometer and clean the pipeline with ultrapure water for 10 min.

[0085] 3. Load the samples onto the flow cytometer in the order listed above. Clean the pipeline with PBS solution after each sample is tested. Use PBS solution to perform blank correction. Control 1 and control 2 are used as negative controls.

[0086] 4. After testing is complete, turn off the flow cytometer and analyze the experimental data.

[0087] Seven, high-throughput sequencing sample preparation and sequencing results:

[0088] 1. Add single-stranded template (library after each round of screening) to 400 μL of PCR mixture solution according to (0.5 μM * 10 μL) and divide into eight PCR tubes, 100 μL per well. Perform PCR amplification for 25 cycles using a Bio-Rad instrument. The PCR program is as follows: 95°C pre-denaturation for 1 min; 95°C for 60 s, 60°C for 60 s, and 72°C for 60 s for 25 cycles.

[0089] 3. After PCR is complete, take 20 μL of each sample for storage and electrophoresis detection. The remaining mixture is divided into two 15 mL centrifuge tubes and concentrated with n-butanol to a volume of about 100 μL.

[0090] 4. Use the μNIQ-10 oligonucleotide purification kit to recover the double-stranded DNA.

[0091] Eight, sequencing result arrangement and analysis

[0092] 1. Extract the sequences using the analysis software according to the tag primer from the obtained raw sequencing results, a total of 1048576 sequences. Note: The extracted results are random sequences, i.e., the sequences of the two end primers are removed. When synthesizing the aptamer, the primer region sequences need to be added before and after the extracted sequences.

[0093] 2. Analyze the obtained sequences, select the sequences to be sequenced, and synthesize monoclonal antibodies. The library is a closed-loop library, and when synthesizing monoclonal antibodies, the open-loop region at both ends of the library is removed. The sequencing results are obtained and ranked and sorted according to the sequence richness.

[0094] Nine, the verification of specific binding KIM-1 DNA aptamer:

[0095] According to the abundance of nucleic acid sequence ranking, then entrusts the biological technology limited company to synthesize 5' label fluorescent FAM nucleic acid aptamer to carry on the subsequent verification work, for example, the first 10 nucleic acid sequences as shown in the following table 4:

[0096] Table 4

[0097]

[0098] In order to reduce the workload and unnecessary waste, screening high binding force ssDNA to do further dissociation constant determination:

[0099] 1. According to the screening sequencing results, the top 10 sequences were synthesized, and the FAM labeled ssDNA nucleic acid aptamer was synthesized.

[0100] 2. 100 pM of FAM labeled nucleic acid aptamer was mixed gently with 200 μL of PBS solution containing 1 × 10 6 Target cells, incubated at 37℃ for 30 min in the dark, and 20 μL of 3% BSA was added; each group was set up three repeat groups.

[0101] 3. 4℃, 350g centrifugation for 5min, discard the supernatant, and wash twice with 2-3mL PBS to remove the unbound and weakly bound ssDNA.

[0102] 4. 4℃, 350g centrifugation for 5min, discard the supernatant, resuspend the cell pellet with 200 μL of pre-cooled PBS, and filter the cells with filter membrane and collect into flow tube.

[0103] 5. The fluorescence intensity of nucleic acid aptamer binding was detected by flow cytometry, and the data obtained by FCM were statistically analyzed to sort the 10 ssDNA sequences.

[0104] The results are shown in Figure 3 It can be seen that K1-K10 can effectively bind to target cells, which can be used for flow cytometry detection of target cells or screening of target cells. According to the method, K11-K70 is screened, and the nucleic acid with strong binding force is further detected for dissociation constant.

[0105] Ten, flow cytometry (FCM) determination of dissociation constant of single nucleic acid aptamer

[0106] 1. According to the results of step nine, select nucleic acid aptamer with strong binding force for dissociation constant determination, the method is as follows:

[0107] 2. Set different concentrations of FAM-labeled aptamer (0, 0.25, 0.50, 1.0, 1.25, 1.50, 1.75, 2.0, 2.25 μM nine ssDNA concentration gradients), and 1 x 10 6 HK-2 cells were incubated at 37°C in the dark for 30 min, and 20 μL of 3% w / v BSA solution was added (three replicate groups were set for each group).

[0108] 3. Centrifuge at 350g for 5 min at 4°C, discard the supernatant, and wash twice with 2-3 mL of PBS to remove unbound and weakly bound ssDNA.

[0109] 4. Centrifuge at 350g for 5 min at 4°C, try to remove the supernatant, resuspend the cell pellet with 200 μL of pre-cooled PBS, and filter the cells with a filter membrane and collect them into a flow tube.

[0110] 5. Use a flow cytometer to detect the fluorescence intensity of aptamer binding, and statistically analyze the data obtained by FCM to calculate the dissociation constant (Kd value) of aptamer.

[0111] The results are shown in Figure 4 The binding rate of aptamer to cells increased with the increase of aptamer concentration, and reached a plateau at a certain concentration, and then did not increase. According to formula (1) and GraPHPad Prism 9.0, the dissociation constant Kd value of aptamer was calculated.

[0112] Y = Bmax X / (Kd+X) (1);

[0113] Where Y represents the average absorbance value, Bmax represents the maximum absorbance value measured, Kd is the dissociation constant of aptamer, and X is the concentration of individual aptamer.

[0114] In the detection of single aptamer binding capacity, as Figure 5 shown: FAM-labeled aptamer K3 and K34 have good affinity to target cells (Kd value reaches nM level), indicating that the aptamer has high affinity to KIM-1 cells.

[0115] Eleven. Fluorescence microscope observation of Cy3-labeled aptamer on KIM-1 cells

[0116] 1. According to the previous screening results, Cy3-labeled ssDNA aptamer Cy3-K3 and Cy3-K34 were synthesized by Genescript Biotech Co., Ltd.

[0117] 2. 3 x 10 5HK-2 cells were seeded into 12-well cell culture plates and divided into groups 1, 2, 3, and 4, with 3 wells in each group. Groups 1 and 2 were treated with culture medium containing 30 μg / mL vancomycin for 24 hours, while groups 3 and 4 were treated with PBS as a control.

[0118] 3. After 24 hours, wash three times with PBS. Add 200 μL of PBS containing 100 pM Cy3-K3 to groups 1 and 3 and incubate in the dark for 30 min. Add 200 μL of PBS containing 100 pM Cy3-K34 to groups 2 and incubate in the dark for 30 min.

[0119] 4. After incubation, wash three times with PBS and add 100μL of PBS to keep moist.

[0120] 5. Use a fluorescence microscope for observation and photography.

[0121] The results are as follows Figure 6 As shown, the Cy3-labeled nucleic acid aptamers we screened can be used to label target cells for fluorescence microscopy detection.

[0122] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A nucleic acid aptamer that binds to KIM-1 with high affinity, characterized in that: The nucleic acid aptamer targeting KIM-1 is at least one of a nucleic acid aptamer K3 with a nucleic acid sequence as shown in SEQ ID NO. 1 and a nucleic acid aptamer K34 with a nucleic acid sequence as shown in SEQ ID NO.

2.

2. The nucleic acid aptamer targeted to bind KIM-1 according to claim 1, characterized in that: The nucleic acid aptamer targeting KIM-1 is a labeled nucleic acid aptamer.

3. The nucleic acid aptamer targeting KIM-1 according to claim 2, wherein: The labeled substance is at least one of colloidal gold, biotin and a fluorescent molecule.

4. The nucleic acid aptamer that binds KIM-1 according to claim 3, characterized in that: The fluorescent molecule is at least one of FAM and Cy3.

5. Use of the nucleic acid aptamer targeting KIM-1 according to any one of claims 1 to 4 in the preparation of a KIM-1 detection reagent.

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