CN112415206B - Application of CD171 protein in exosome as tumor metastasis diagnosis marker - Google Patents
Application of CD171 protein in exosome as tumor metastasis diagnosis marker Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57496—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving intracellular compounds
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
- G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
Abstract
The invention relates to the field of medical diagnostics, in particular to application of CD171 protein in exosomes as a tumor metastasis diagnosis marker. The present invention provides a method for predicting tumor metastasis and its metastatic sites. The CD171 protein in the exosomes specifically related to the invention can be used as a tumor metastasis prediction marker, and detection of the CD171 protein in the exosomes from a tissue-specific source can be used for predicting tumor metastasis sites. The method provided by the invention can be applied to the prediction of tumor metastasis, and also provides an important reference for tumor prognosis evaluation and the selection of tumor treatment schemes.
Description
Technical Field
The invention relates to the field of medical diagnostics, in particular to application of CD171 protein in exosomes as a tumor metastasis diagnosis marker.
Background
Tumor occurrence is a multifactorial, multi-stage evolutionary process. In recent years, the expression level of CD171 in cancer and the metastasis capability are positively correlated, and CD171 protein can increase the migration of tumor cells. Metastatic tumor cells lacking CD171 can reach and drill capillaries through the blood circulation, but lose the ability to migrate along the outer wall of the blood vessel and grow into tumors. Normal cancer cells move along capillaries and can squeeze off perivascular cells along the way, but after CD171 is knocked out, cancer cells do not squeeze off perivascular cells; in contrast, in a cancer cell that underexpresses CD171, over-expressing CD171 increases the ability of the cancer cell to migrate along the vascular network. CD171 protein is critical for tumor implantation.
There is a strong need in the art for methods that allow for the prediction of effective tumor metastasis, prognosis evaluation of tumors, and selection of tumor treatment regimens.
Disclosure of Invention
The invention relates to an application of a detection agent in preparation of a kit;
the detection agent comprises a quantitative detection agent of CD171 protein in exosomes;
the kit is used for predicting whether the tumor in the subject is metastasized, predicting the metastatic site, judging the prognosis of the tumor treatment and selecting at least one of tumor treatment schemes;
the exosomes are derived from tissue-specific exosomes in a biological sample;
the tissue-specific exosome is at least one selected from intestinal tissue-specific exosome, gastric tissue-specific exosome, liver tissue-specific exosome, lung tissue-specific exosome, and lymphoid tissue-specific exosome.
Optionally, for use as described above, the detection agent further comprises at least one qualitative detection agent selected from the group consisting of:
intestinal tissue specific marker MS4a12, gastric tissue specific marker SLC9A4, liver tissue specific marker SLCO1B3, lung tissue specific marker agrer, and lymphoid tissue specific marker CD8A.
Optionally, the use of the detection agent as described above is for performing any one of the following methods:
biological mass spectrometry, electrophoresis, chromatography, enzyme-linked immunosorbent assay, immunofluorescence, immunochromatography, immunonephelometry, immunoblotting and dot blotting.
Alternatively, for use as described above, the detection agent is a specific antibody directed against the substance to be detected.
Optionally, the biological sample is selected from at least one of cell culture supernatant, whole blood, serum, plasma, ascites, cerebrospinal fluid, bone marrow aspirate, bronchoalveolar lavage, urine, semen, vaginal secretion, mucus, saliva, sputum, and clarified lysate obtained from a biological tissue sample, for use as described above.
The invention also contemplates a kit comprising an exosome enrichment purification reagent and a detection reagent as defined above;
the kit is used for at least one of predicting whether a tumor in a subject metastasizes, predicting a metastatic site, prognosis of tumor treatment, and selecting a tumor treatment regimen.
Optionally, the kit as described above further comprises at least one of a solid support, a protease inhibitor, a blocking solution, a color reagent, and a wash buffer.
Alternatively, the kit as described above, the solid support is a chemiluminescent plate.
The invention also relates to a system, which comprises an information acquisition module and a data analysis reporting module;
the information acquisition module has the functions of: acquiring the concentration of CD171 protein in exosomes and providing the results to the data analysis module;
the functions of the data analysis reporting module include: judging whether the concentration is more than or equal to 150pg/mL, if so, outputting that the risk of tumor metastasis is high; if not, the output is low in risk of tumor metastasis.
The beneficial effects of the invention are as follows:
the present invention provides a method for predicting tumor metastasis and its metastatic sites. The CD171 protein in the exosomes specifically related to the invention can be used as a tumor metastasis prediction marker, and detection of the CD171 protein in the exosomes from a tissue-specific source can be used for predicting tumor metastasis sites. The method provided by the invention can be applied to the prediction of tumor metastasis, and also provides an important reference for tumor prognosis evaluation and the selection of tumor treatment schemes.
Drawings
In order to more clearly illustrate the embodiments of the present invention 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 invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the results of total exosome CD171 protein detection in metastatic and non-metastatic cancer samples according to one embodiment of the present invention;
FIG. 2 shows the results of detecting CD171 protein of tissue-specific source exosomes of a sample of a patient with liver metastasis from gastric cancer in one embodiment of the present invention;
FIG. 3 shows the results of detecting CD171 protein of exosomes of specific origin in a sample of patients with liver metastasis of intestinal cancer according to one embodiment of the present invention;
FIG. 4 shows the results of detecting CD171 protein of tissue-specific source exosomes of liver cancer lung metastasis patient samples according to one embodiment of the present invention.
Detailed Description
Reference now will be made in detail to embodiments of the invention, one or more examples of which are described below. Each example is provided by way of explanation, not limitation, of the invention. Indeed, it will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the scope or spirit of the invention. For example, features illustrated or described as part of one embodiment can be used on another embodiment to yield still a further embodiment.
Accordingly, it is intended that the present invention cover such modifications and variations as fall within the scope of the appended claims and their equivalents. Other objects, features and aspects of the present invention will be disclosed in or be apparent from the following detailed description. It is to be understood by one of ordinary skill in the art that the present discussion is a description of exemplary embodiments only, and is not intended as limiting the broader aspects of the present invention.
The invention relates to an application of a detection agent in preparation of a kit;
the detection agent comprises a quantitative detection agent of CD171 protein in exosomes;
the kit is used for predicting whether the tumor in the subject is metastasized, predicting the metastatic site, judging the prognosis of the tumor treatment and selecting at least one of tumor treatment schemes;
the exosomes are derived from tissue-specific exosomes in a biological sample;
the tissue-specific exosome is at least one selected from intestinal tissue-specific exosome, gastric tissue-specific exosome, liver tissue-specific exosome, lung tissue-specific exosome, and lymphoid tissue-specific exosome.
The present invention provides a novel marker for diagnosis: CD171 protein in exosomes.
Wherein an elevated level of CD171 protein in the subject's exosomes as compared to a control level from a subject without the tumor metastasis indicates that the subject is likely to have tumor metastasis, and an elevated level of CD171 in the tissue-specific source exosomes in the subject indicates a corresponding tumor metastasis site.
The term "marker" as used herein refers to a molecule to be used as a target for analysis of a patient experimental sample. Examples of such molecular targets are proteins or polypeptides. Proteins or polypeptides used as markers in the present invention are intended to include naturally occurring variants of said proteins as well as fragments, in particular immunologically detectable fragments, of said proteins or said variants. The immunologically detectable fragment preferably comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 15 or 20 consecutive amino acids of the marker polypeptide. Those skilled in the art will recognize that proteins released by cells or present in the extracellular matrix may be damaged (e.g., during inflammation) and may be degraded or cleaved into such fragments. Certain markers are synthesized in inactive form, which can be subsequently activated by proteolysis. As will be appreciated by the skilled artisan, the protein or fragment thereof may also be present as part of a complex. Such complexes can also be used as markers in the sense of the present invention. In addition, or in the alternative, the marker polypeptide or variant thereof may carry a post-translational modification. Non-limiting examples of post-translational modifications are glycosylation, acylation, and/or phosphorylation.
In some embodiments, the tumor is a solid tumor.
In some embodiments, the quantitative detection agent further comprises at least one detection agent selected from the group consisting of:
intestinal tissue specific marker MS4a12, gastric tissue specific marker SLC9A4, liver tissue specific marker SLCO1B3, lung tissue specific marker agrer, and lymphoid tissue specific marker CD8A.
In some embodiments, the quantitative detection agent is used to perform any one of the following methods:
biological mass spectrometry, electrophoresis, chromatography, enzyme-linked immunosorbent assay, immunofluorescence, immunochromatography, immunonephelometry, immunoblotting and dot blotting.
Quantitative detection agents are typically reagents that specifically detect CD171 protein in exosomes and optionally tissue-specific markers, e.g., lectins that specifically bind to one of them, aptamers that specifically bind to one of them, or antibodies and antibody fragments that specifically bind to one of them. Specific binding agents having at least 10 for their corresponding target molecules 7 l/mol affinity. The specific binding agent preferably has a binding agent of 10 for its target molecule 8 l/mol, or more preferably 10 9 l/mol affinity.
In some embodiments, the quantitative detection agent is a specific antibody directed against the analyte.
In some embodiments, the specific antibody is a monoclonal antibody or a polyclonal antibody.
In some embodiments, the biological sample is selected from at least one of cell culture supernatant, whole blood, serum, plasma, ascites, cerebrospinal fluid, bone marrow aspirate, bronchoalveolar lavage, urine, semen, vaginal secretion, mucus, saliva, sputum, and clarified lysate obtained from a biological tissue sample.
The invention also provides a kit comprising an exosome enrichment purification reagent and a quantitative detection reagent as defined above;
the kit is used for at least one of predicting whether a tumor in a subject metastasizes, predicting a metastatic site, prognosis of tumor treatment, and selecting a tumor treatment regimen.
In some embodiments, the kit further comprises at least one of a solid support, a protease inhibitor, a blocking solution, a color reagent, and a wash buffer.
The protease inhibitor can be one or more of Indinavir, saquinavir, ritonavir, nelfinavir, amprenavir.
The blocking solution can be one or more of BSA, bovine serum, skimmed milk, TBST, etc.
The color development solution can be determined based on the substance labeled on the antibody, for example, when the labeled substance is horseradish peroxidase, the color development agent can be luminol.
The wash buffer may be PBS, TBS, or the like.
The protease inhibitor, the blocking solution, the color development solution and the washing buffer may be packaged in the form of working concentrations in the kit, or may be packaged in the form of concentrated mother solutions thereof (e.g., mother solutions concentrated 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, and 50 times).
The solid phase carrier is usually used for coating the antibody, and the solid phase carrier material used for coating the antibody can be polystyrene, cellulose, polyacrylamide, polyethylene polypropylene, sephadex, glass, silicone rubber, agarose gel and other materials, and the carrier can be in the form of a test tube, an EP tube, a porous plate (especially a chemiluminescent plate), a concave hole of a micro-reaction plate, a small bead (especially a magnetic bead), a small disc and the like.
The preferred solid support is a chemiluminescent plate. It may contain holes of 16, 32, 48, 64, 96 or more.
According to yet another aspect of the present invention, there is also provided a system including an information acquisition module and a data analysis reporting module;
the information acquisition module has the functions of: acquiring the concentration of CD171 protein in exosomes and providing the results to the data analysis module;
the functions of the data analysis reporting module include: judging whether the concentration is more than or equal to 150pg/mL, if so, outputting that the risk of tumor metastasis is high; if not, the output is low in risk of tumor metastasis.
In some embodiments, the exosomes are derived from a cancer tissue (designated as a) that has been diagnosed.
In some embodiments, the exosomes may also be derived from tissue (denoted B) in which metastasis may occur; further, if the concentration of CD171 protein of B is not less than 150pg/mL, it is indicated that the risk of metastasis of tumor cells of A to B is high.
The invention also relates to a method of diagnosing whether a tumor metastasizes, the method comprising:
a) Enriching and purifying exosomes;
b) Detecting the concentration of CD171 protein in the exosomes;
c) If the concentration is not less than 150pg/mL, the probability of transition is high.
In some embodiments, the enriched exosomes are isolated from the subject biological sample in step a) according to the principle of molecular sieve chromatography (e.g. using CL-6B packing).
In some embodiments, step b) simultaneously detects a marker of at least one of an intestinal tissue-specific exosome, a gastric tissue-specific exosome, a liver tissue-specific exosome, a lung tissue-specific exosome, a lymphoid tissue-specific exosome.
In some embodiments, the marker is selected from at least one of the following: intestinal tissue specific marker MS4a12, gastric tissue specific marker SLC9A4, liver tissue specific marker SLCO1B3, lung tissue specific marker agrer, and lymphoid tissue specific marker CD8A.
The subject is typically a mammal, preferably a primate, more preferably a human.
It should be noted that the ideal scenario for diagnosis is a situation in which a single event or process causes various diseases. In all other cases, correct diagnosis can be very difficult, especially when the etiology of the disease is not fully understood, as in the case of many cancer types. As will be appreciated by the skilled artisan, for a given multifactorial disease, diagnosis without biochemical markers is 100% specific and as sensitive as 100%. Conversely, biochemical markers (e.g., CD171 protein concentration in exosomes as demonstrated herein) may be used to assess, for example, the presence or absence or severity of a disease with a certain probability or predictive value. Thus, in routine clinical diagnosis, various clinical symptoms and biological markers are often taken into consideration in combination to diagnose, treat and manage underlying diseases.
Embodiments of the present invention will be described in detail below with reference to examples.
EXAMPLE 1 isolation of exosomes of serum tissue specific origin
Plasma was removed from the-80 ℃ freezer and 2mL of each plasma sample was quickly thawed before 0.6mL of thromboplastin-D was added and incubated for 60 minutes at room temperature. 1.4mL of phosphate buffer containing a protease inhibitor cocktail was added. Centrifuge at 3000g for 20 min at 4 ℃. After centrifugation, the supernatant was taken and the enriched exosomes were isolated using CL-6B packing and concentrated to a volume of 2mL. 200uL of each exosome solution was taken, 2uL of biotinylated antibody to the tissue specific marker was added, and 50uL of 3% BSA was added and placed in a refrigerator at 4℃for 1 hour. After that, 25uL of streptavidin agarose was added, and 50uL of 3% BSA was added thereto, and the mixture was left at 4℃for 30 minutes. Centrifugation at 400g for 10 min at 4℃was performed to remove the supernatant thoroughly, leaving the pellet. The precipitate was added with 50uL of 0.05M glycine-hydrochloric acid (ph=3), vortexing for 10 seconds. Resuspended tissue-specific derived exosome particles, stored in a-80℃refrigerator,
example 2 determination of exosome expression CD171 protein levels
The extracted exosome samples were removed from the-80℃refrigerator, thawed quickly, added with 0.5mL M-PER, and the pH of the solution was adjusted to 8.0 with 1M Tris-HCl pH 8.6. Incubated in a constant temperature water bath at 37 ℃ for 10 minutes, and vortexed for 10 seconds to completely lyse exosomes. And taking out the CD171 detection kit from the refrigerator, balancing to room temperature, respectively adding 100uL of calibrator and sample to be detected into the sample hole to be detected, and setting a compound hole. Incubate at 37℃for 1 hour. After washing the plate 3 times, 100uL of detection antibody was added and incubated in an incubator at 37℃for 1 hour. After washing the plate 3 times, 100uL of chemiluminescent substrate was added, and the luminescence value was read by a chemiluminescent instrument. And drawing a calibration curve according to the luminous value of the calibrator, and calculating the concentration of the sample from the calibration curve.
Example 3 Total exosome CD171 protein detection in metastatic and non-metastatic cancer samples
Collecting 50 cases of cancer metastasis plasma samples, including 10 cases of gastric cancer samples, 10 cases of intestinal cancer samples, 10 cases of lung cancer samples and 10 cases of liver cancer samples; and collecting cancer non-metastatic plasma samples, wherein the cancer non-metastatic plasma samples comprise 10 gastric cancer samples, 10 intestinal cancer samples, 10 lung cancer samples and 10 liver cancer samples. After isolation of the enriched exosomes, the surface marker CD171 protein expression level was cleaved and detected. As can be seen from FIG. 1, the CD171 expression level of the non-metastatic cancer sample is mostly lower than 150pg/mL, and the CD171 expression level of the metastatic cancer sample is mostly higher than 150pg/mL.
TABLE 1 Total exosome CD171 protein detection for metastatic and non-metastatic cancer samples
Example 4 detection of CD171 protein in tissue-specific Source exosomes prediction of tumor metastasis sites
Plasma samples of 30 metastatic cancer patients were collected, including 10 patients with gastric cancer liver metastasis, 10 patients with intestinal cancer liver metastasis, and 10 patients with liver cancer lung metastasis. Exosomes of tissue-specific origin were isolated and captured, and the expression level of the surface marker CD171 protein was detected. FIG. 2 shows the results of detecting tissue-specific source exosome CD171 protein in gastric cancer liver metastasis patient, and it can be seen from the graph that the expression level of liver tissue-source exosome CD171 protein is obviously higher than that of other tissues. FIG. 3 shows the results of detecting tissue-specific source exosome CD171 protein in intestinal cancer liver metastasis patient, and the expression level of liver tissue-source exosome CD171 protein is obviously higher than that of other tissue. FIG. 4 shows the result of detecting the CD171 protein of the exosome of the specific source of the sample tissue of the liver cancer lung metastasis patient, and the expression level of the CD171 protein of the exosome of the lung tissue source is obviously higher than that of other tissues.
TABLE 2 detection of tissue-specific exosome CD171 protein in gastric cancer liver metastasis patient samples
TABLE 3 detection of tissue-specific exosome CD171 protein in sample of patient with intestinal cancer liver metastasis
TABLE 4 detection of tissue-specific exosome CD171 protein in liver cancer lung metastasis patient samples
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (10)
1. The application of the detection agent in the preparation of the kit,
the detection agent comprises a quantitative detection agent of CD171 protein in exosomes;
the kit is used for predicting at least one of tumor metastasis sites, prognosis of tumor treatment based on the tumor metastasis sites, and selecting a tumor treatment regimen based on the tumor metastasis sites;
the exosomes are derived from tissue-specific exosomes in a biological sample;
the tissue-specific exosome is selected from at least one of intestinal tissue-specific exosome, gastric tissue-specific exosome, liver tissue-specific exosome, lung tissue-specific exosome, and lymphoid tissue-specific exosome;
the tumor is at least one selected from intestinal cancer, liver cancer, gastric cancer and lung cancer;
the tumor transfer part is selected from at least one of intestinal cancer tissue, liver cancer tissue, gastric cancer tissue, lung cancer tissue and lymph tissue;
the biological sample includes a plasma sample.
2. The use according to claim 1, wherein the detection agent further comprises at least one qualitative detection agent selected from the group consisting of tissue-specific markers:
intestinal tissue-specific markers, gastric tissue-specific markers, liver tissue-specific markers, lung tissue-specific markers, and lymphoid tissue-specific markers.
3. The use according to claim 1 or 2, wherein the detection agent further comprises at least one qualitative detection agent selected from the group consisting of tissue-specific markers:
intestinal tissue specific marker MS4a12, gastric tissue specific marker SLC9A4, liver tissue specific marker SLCO1B3, lung tissue specific marker agrer, and lymphoid tissue specific marker CD8A.
4. The use according to any one of claims 1 to 3, wherein the detection agent is used to perform any one of the following methods:
biological mass spectrometry, electrophoresis, immunofluorescence, immunonephelometry, and dot blotting.
5. The use according to claim 4, wherein the detection agent is a specific antibody against the substance to be detected.
6. The use according to claim 1, wherein the tumour is a solid tumour.
7. The use according to any one of claims 1 to 6, wherein the kit further comprises a total exosome enrichment reagent.
8. The use according to claim 1, wherein the kit further comprises at least one of a solid support, a protease inhibitor, a blocking solution, a chromogenic agent and a wash buffer.
9. The use according to claim 8, wherein the solid support is a chemiluminescent plate.
10. A system comprising an information acquisition module and a data analysis reporting module;
the information acquisition module has the functions of: obtaining the concentration of CD171 protein in an exosome from a tissue-specific exosome in a biological sample, the biological sample comprising a plasma sample, and providing the result to the data analysis reporting module;
the tissue-specific exosome is selected from at least one of intestinal tissue-specific exosome, gastric tissue-specific exosome, liver tissue-specific exosome, lung tissue-specific exosome, and lymphoid tissue-specific exosome;
the functions of the data analysis reporting module include: and predicting a tumor metastasis site according to the concentration and the tissue corresponding to the exosome, wherein the tumor is selected from at least one of intestinal cancer, liver cancer, gastric cancer and lung cancer, and the tumor metastasis site is selected from at least one of intestinal cancer tissue, liver cancer tissue, gastric cancer tissue, lung cancer tissue and lymph tissue.
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