196 results about "Immunoadsorption" patented technology

The invention relates to the technical field of biology, in particular to an anti-B7-H3 monoclonal antibody, a hybridoma cell strain and application thereof. The invention provides a monoclonal antibody. Amino acid sequences of three CDR regions of a heavy chain of the monoclonal antibody respectively have amino acid sequences shown as SEQ ID NO: 11, 12 and 13; the amino acid sequences of the three CDR regions of the light chain of the gene respectively have the amino acid sequences shown as SEQ ID NO: 6, 7 and 8. The anti-B7-H3 monoclonal antibody provided by the invention can be used for specifically recognizing a B7-H3 recombinant protein expressed by eukaryotic expression and a B7-H3 protein expressed on a cell membrane. A B7-H3 detection kit constructed by the anti-B7-H3 monoclonal antibody is high in sensitivity and good in specificity, and has a better application effect in various detections including enzyme-linked immunosorbent assay, flow cytometry, immunofluorescence and the like.

The invention discloses an anti-Ebola virus VP40 protein nano antibody and an application of the anti-Ebola virus VP40 protein nano antibody in Ebola virus detection. The anti-Ebola virus VP40 protein nano antibody comprises a 3E07 antibody and a 3F06 antibody. The anti-Ebola virus VP40 protein nano antibody prepared by the invention has the characteristics of high sensitivity, wide binding spectrum and the like. A double-antibody sandwich enzyme-linked immunosorbent assay method constructed by the 3E07 and 3F06 antibodies can be used for efficiently detecting VP40 proteins and virus-like particles of various subtype Ebola viruses such as Zaire, Sudan, Bandong and the like, and the detection sensitivity for the VP40 proteins of the Zaire type Ebola viruses can reach 0.039 ng / mL; the antibody combination has a wide application prospect in diagnosis and pathogen detection of Ebola virus diseases.

The invention belongs to the field of immunological detection, and provides a double-antibody sandwich ELISA kit for detecting the content of human beta-NGF. The enzyme-linked immunosorbent assay kit constructed by the invention is based on a double-antibody sandwich method, and is combined with a biotin-streptavidin signal amplification system and an enzyme developing amplification system, so that the detection sensitivity and specificity are greatly improved, and low-content samples can be accurately and quickly detected. The kit shows excellent stability and specificity and can be used for quantitatively detecting the content of beta-NGF in samples such as human serum, cell supernatant or tissue homogenate, so that an objective detection basis is provided for diagnosis, curative effect evaluation and pathogenesis research of related diseases.

The invention relates to a hybridoma cell strain VAND8, an anti-vanillin monoclonal antibody generated by the hybridoma cell strain VAND8 and application, and belongs to the field of immunochemistry. The hybridoma cell strain VAND8 provided by the invention can be used for preparing a high-titer vanillin-resistant monoclonal antibody, and the titer measured by an enzyme-linked immunosorbent assay (ELISA) method can reach 3.2 * 10 < 5 >; the vanillin-resistant monoclonal antibody provided by the invention is high in sensitivity and good in specificity, the 50% inhibition concentration IC50 on vanillin is 22.1 mu g / mL, and the cross reaction rate with vanillin structural analogues is low or even no cross reaction exists; the method can be used for measuring the content of vanillin.

The invention relates to a porcine reproductive and respiratory syndrome virus (PRRSV) dimer nucleocapsid (N) protein (PRRSV N-dimer) and application thereof. The PRRSV N-dimer is formed by connecting N protein amino acid sequences in series through (GGGGS) 3linker. According to the invention, a GGGGS 3linker nucleotide sequence is connected in series with a PRRSV NADC30-like strain N protein gene sequence to construct a prokaryotic expression vector, and a recombinant PRRSV N-dimer is expressed in escherichia coli. Through SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and Western-blot detection, the recombinant PRRSV N-dimer obtains soluble expression and can be subjected to a specific reaction with PRRSV positive serum; through detection by a freeze thawing method and an ELISA (Enzyme-Linked Immunosorbent Assay) method, compared with a PRRSV monomer N protein (PRRSV N-monomer), the PRRSV N-dimer protein can be stably preserved, and the immunoreactivity is better. A PRRSV antibody indirect enzyme-linked immunosorbent assay (iELISA) detection method is established on the basis of a PRRSV N-dimer, and can be applied to diagnosis and monitoring of PRRSV infection.

The invention discloses a KLHL11 antibody high-precision detection method based on an enzyme-linked immunosorbent assay. The detection method comprises the following steps: preparing a microwell plate pre-coated with an anti-human KLHL11 antibody, dividing micropores into a plurality of detection groups, and adding KLHL11 protein or peptide fragment solutions with different concentrations, a to-be-detected solution with unknown concentration and a contrast solution into each detection group. The detection precision of a current detection instrument, the operation capability level of an operator and the reliability of the current detection instrument and the operation capability level are evaluated by executing enzyme-linked immunosorbent assay operation steps including sample filling, incubation, washing, color development and optical density measurement, obtaining the optical density value of a detection group and calculating the mean value, the standard deviation and the variation index of each group. By establishing a standard curve of optical density and concentration, the concentration of an unknown sample can be calculated, and the performance of an instrument in different concentration ranges can be verified. In addition, the problems of drifting and cross contamination of the instrument are further detected by setting a detection group with a relatively large time interval and a high-low concentration gradient experiment.

The invention discloses a novel diagnostic kit for evaluating the autophagy function of a mitochondrial network, which is used for detecting the content of autophagy related proteins NADH, ROMO1, PINK1 and TOM40 of four mitochondrial networks of a human body sample by utilizing an enzyme-linked immunosorbent assay (ELISA) and comprehensively judging the functional state of the mitochondrial of the human body. According to the kit, a microplate reader is used for measuring the absorbance (OD value) of a sample, and the concentration of the sample is calculated through a standard curve. Results show that the kit can accurately detect the content of four proteins, comprehensively judge the mitochondrial network autophagy ability of a human body sample and understand the mitochondrial function state. The kit for diagnosing the mitochondrial function of the double-antibody sandwich method combined with four proteins can conveniently and accurately evaluate the autophagy ability of the mitochondrial network in a human body specimen. The invention discloses preparation of a diagnostic kit for evaluating the autophagy ability of a mitochondrial network. The diagnostic kit can be used for evaluating cell mitochondrial states in cell mitochondrial functions, related disease diagnosis and treatment indexes and scientific research.

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The invention discloses a high-affinity monoclonal antibody for recognizing group B streptococcus SIP (Session Initiation Protocol) protein or an antigen binding fragment thereof. The amino acid sequence of a heavy chain variable region of the antibody is as shown in SEQ ID NO.1, and the amino acid sequence of a light chain variable region of the antibody is as shown in SEQ ID NO.5. The lowest detection limit of the group B streptococcus SIP protein double-antibody sandwich enzyme-linked immunosorbent assay established by taking the monoclonal antibody or the antigen binding fragment thereof as a capture antibody on the SIP protein is 0.32 ng / mL. The lowest detection limit of a group B streptococcus SIP protein double-antibody sandwich fluorescence immunochromatography assay method established by taking the monoclonal antibody or the antigen binding fragment thereof as a capture antibody on SIP protein is 0.08 ng / mL, and the lowest detection limit on GBS bacterial liquid reaches 1 * 10 < 3 > CFU / mL, which is obviously higher than the lowest detection limit of the existing detection reagent and literature report; the method can be used for qualitative and quantitative detection of the group B streptococcus SIP protein.

1. The name of the design product: Fully automatic enzyme-linked immunosorbent assay workstation. 2. The use of the design product: For enzyme-linked immunosorbent assay or detection. 3. The design key points of the design product: Lies in the shape. 4. The picture or photo that best shows the design key points: Perspective view.

The invention discloses a paralichthys olivaceus rhabdovirus G protein tandem antigen epitope peptide and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO: 1. The preparation method comprises the following steps: (1) firstly, analyzing structural characteristics of HIRRV-G protein, predicting B cell antigen epitopes of the HIRRV-G protein, screening the antigen epitopes with advantages on the basis of a prediction result, and synthesizing the antigen epitopes; (2) screening a candidate peptide fragment with high affinity through an enzyme-linked immunosorbent assay; and (3) sequentially connecting the high-affinity peptide fragment sequences meeting the requirements by using a GPGPG connexon, cloning the connected sequences into a pET-28a prokaryotic expression vector, and performing induced expression to obtain the tandem antigen epitope peptide. Compared with a full-length G protein, the tandem antigen epitope peptide is smaller in molecular weight, higher in stability and higher in hydrophilicity; a large number of specific antibodies can be induced in fish bodies, and the death rate of the paralichthys olivaceus infected by viruses is remarkably reduced. The method can be used for HIRRV diagnosis detection reagent and subunit vaccine development.

The invention discloses a detection method based on combination of anti-NF155 antibody detection and enzyme-linked immunosorbent assay. According to the detection method, an analysis model based on an XGBoost model algorithm is established by adopting anti-NF155 antibody detection of a patient and detection data of an enzyme-linked immunosorbent assay in combination with autologous clinical data of the patient, so that CIDP subtype classification is performed on the patient, and personalized treatment scheme suggestions are provided in combination with illness state dynamic data of the patient. Meanwhile, the invention provides a corresponding detection system which comprises a laboratory, a data storage library and an analysis unit, the anti-NF155 antibody detection is executed through laboratory equipment, and detection data and patient autologous clinical data are stored in the data storage library. The analysis unit establishes an analysis model based on an XGBoost model algorithm based on a machine learning algorithm.

The invention provides a human metapneumovirus monoclonal antibody and application thereof, and belongs to the technical field of immunity. The monoclonal antibody F720-1 provided by the invention only has binding activity to conformation F protein before human metapneumovirus fusion, and the monoclonal antibody F390-6 has binding activity to conformation F protein before human metapneumovirus fusion and conformation F protein after human metapneumovirus fusion. Through pairing, it is determined that F720-1 and F390-6 can conduct proportion detection on Pre-F and Post-F in the human metapneumovirus F protein through an enzyme-linked immunosorbent assay double-antibody sandwich method and conduct quantification on Pre-F. The double-antibody sandwich ELISA method has a good linear relation and detection limit when used for detecting the proportion of the Pre-F protein to the Post-F protein in the F protein. The method is expected to be used for accurate evaluation in human metapneumovirus protein experimental vaccine stock solution and vaccine verification.

The invention discloses application of hydrogen-rich water in preparation of a drug for treating Duchenne type muscular dystrophy, and particularly relates to the technical field of medicines.The random control experiment is adopted, homologous c57 mice serve as a normal control group, mdx mice are randomly divided into a model group and a hydrogen-rich water group, and the exercise ability is evaluated through a four-limb holding power test and a rotating bar test; detecting mouse creatine kinase by an enzyme-linked immunosorbent assay; carrying out hematoxylin-eosin, NADH-tetrazole reductase and improved Goori three-color dyeing, and carrying out cytochrome C oxidase, succinate dehydrogenase and SDH-COX combined dyeing to observe the pathological change of the skeletal muscle tissue; the invention discloses that the muscle injury condition of a Duchenne type muscular dystrophy model mouse is improved by the hydrogen-rich water through a mitochondrial protection mechanism, and the hydrogen-rich water has a relatively high application prospect and a relatively high economic value.

The invention discloses an anti-phenacetin monoclonal antibody hybridoma cell strain AD, a preparation method and application thereof, and relates to the technical field of food safety immunodetection. The monoclonal antibody hybridoma cell strain is named monoclonal cell strain AD and the number CGMCC19681. The Phe-BA obtained by the hydrolysis of the reaction product of the phenacetin metabolite acetaminophen and ethyl 4-bromobutyrate is used as the hapten, and the hapten is coupled with the carrier protein to prepare the immunogen Phe-BA-BSA. After the mice were immunized with the immunogen Phe-BA-BSA, they were fused with myeloma cells by PEG method, screened by indirect competitive enzyme-linked immunosorbent assay and subcloned five times to obtain hybridoma cell lines. The monoclonal antibody secreted by the cell line can be made into a phenacetin detection kit, which has good affinity and detection sensitivity for phenacetin, and can be used for immunodetection of phenacetin residues in food.

The application discloses a tandem antigen epitope peptide of G protein of Paralichthys olivaceus rhabdovirus and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO:1. The preparation method of the application comprises the following steps: (1) first, analyzing the structural characteristics of HIRRV-G protein and predicting B cell antigen epitopes, and synthesizing the antigen epitopes with advantages based on the prediction results; (2) then, screening candidate peptide segments with high affinity through enzyme-linked immunosorbent assay; (3) sequentially connecting the high-affinity peptide segment sequences meeting the requirements by using a GPGPG linker, and cloning the connected sequences into a pET-28a prokaryotic expression vector, so that the tandem antigen epitope peptide is obtained after induced expression. Compared with the full-length G protein, the tandem antigen epitope peptide has smaller molecular weight, stronger stability and stronger hydrophilicity; the tandem antigen epitope peptide can induce a large amount of specific antibodies in fish bodies, and significantly reduces the mortality of fish infected by the virus. The tandem antigen epitope peptide can be used for the development of HIRRV diagnostic reagents and subunit vaccines.

Described herein are methods and kits for the detection of platelet-activating pathogenic antibodies in samples from patients suspected of having platelet-activating antibodies or non-HIT anti-PF4 antibody mediated syndromes using platelets and polyvinyl sulfonate or salts thereof. Upon platelet activation, thrombospondin-1 (TSP1), a platelet granule protein, is released and can be detected using a TSP1 antigen capture enzyme linked immunosorbent assay (ELISA) to accurately measure platelet activation.

The application discloses nanobodies against E protein of chikungunya virus and application in virus detection, and the nanobodies against E protein of chikungunya virus include N055 and 10G4 antibodies, and both have high affinity and binding activity with antigens. A double antibody sandwich enzyme-linked immunosorbent assay method constructed by the nanobodies of the N055 and 10G4 antibodies can efficiently detect E protein of chikungunya virus and virus-like particles, and the detection sensitivity can reach 49 pg / mL, and the antibody combination has wide application prospects in diagnosis and pathogen detection of chikungunya virus disease.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention relates to a preparation method of an aluminum hydroxide adjuvant. Specifically, the method comprises the following steps: simultaneously spraying an alkali solution and an aluminum chloride solution into a reaction pipeline in a vaporific form through two pump bodies by virtue of fluid pressure difference, or spraying the alkali solution into a flowing aluminum chloride solution reaction pipeline in a vaporific form by virtue of fluid pressure difference, and controlling the temperature in the pipeline to be 70-80 DEG C for reaction, the reaction liquid flows out through a pipeline within a certain time to obtain an aluminum hydroxide adjuvant crude product, and then the aluminum hydroxide adjuvant is obtained through centrifugation, ion removal, sterilization and packaging. The quality index of the aluminum hydroxide adjuvant obtained by the method is good, stable and uniform, and the immunoadsorption effect of the adjuvant and the quality, effectiveness and safety of a vaccine are ensured. The method is simple in technological process, the whole process is continuously carried out, the barrier of intermittent operation is broken through, operation is simple, and the method can be applied to large-scale production.

In accordance with at least one aspect of this disclosure, a method for preparing a patient for a diagnostic procedure and / or for receiving a therapy using a disease targeted drug is provided. The method can include the steps of a) identifying one or more soluble targets detectable in plasma of the patient; and b) extracorporeally removing the soluble targets from the plasma of the patient. Extracorporeally removing the soluble targets from the plasma of the patient can include using plasmapheresis or immunoadsorption to prepare a patient for a procedure using disease targeted treatment or diagnostics. Additionally, or alternatively, dose modulation can be used to proportionally increase the concertation of the diagnostic / treatment agent relative to the concentration of the soluble target in the blood, improving the outcome treatment and diagnostic procedures over conventional methods.

A method for detecting hepatitis B virus comprising a step of bringing a sample into contact with an antibody which binds to a surface antigen protein of hepatitis B virus, and a lectin. The surface antigen protein of hepatitis B virus may be an S protein, and the lectin may be wheat germ agglutinin. The detection method may involve an ELISA measurement and may be used for testing, prophylaxis, or planning treatment of hepatitis B virus, and for screening medicines.

The invention provides an adsorption column exhaust device and method, and relates to the technical field of blood purification. The adsorption column exhaust device comprises a flushing device, a vacuumizing device and a waste liquid tank, the flushing device is connected to one end of the adsorption column, the vacuumizing device is connected to the other end of the adsorption column, and the vacuumizing device is connected to the waste liquid tank; wherein the vacuumizing device can vacuumize the adsorption column, so that negative pressure is generated in the adsorption column, and bubbles in the adsorption column are expanded under the negative pressure; in the process of vacuumizing the adsorption column, when the pressure in the adsorption column meets the preset pressure condition, the flushing device can convey flushing fluid into the adsorption column, and conveying of the flushing fluid into the adsorption column is stopped after a preset duration; the vacuumizing device can also extract the flushing fluid in the adsorption column and convey the flushing fluid to the waste liquid tank, and meanwhile, the bubbles in the adsorption column are taken away. On the basis, the gas in the adsorption column can be effectively discharged, the adsorption efficiency of the adsorption column on pathogenic substances is improved, and the safety of immunoadsorption is ensured.

The invention provides a method for detecting new antibiotic pollutants in a water body sample by combining solid-phase extraction and enzyme-linked immunosorbent assay, which comprises the following steps: (a) selecting an organic solvent, so that R2 of a standard curve obtained by enzyme-linked immunosorbent assay of a standard substance containing the new antibiotic pollutants and an aqueous solution of the organic solvent is greater than 0.98; (b) determining the optimal concentration of the organic solvent by taking the quantitative detection interval, the sensitivity, the detection limit and the precision of enzyme-linked immunosorbent assay as evaluation indexes and combining a CRITIC weight method; and (c) preparing the new antibiotic pollutants enriched by the organic solvent selected in the step (a) through solid-phase extraction of the water body sample and water into a redissolved aqueous solution, and detecting the redissolved aqueous solution through enzyme-linked immunosorbent assay to determine the concentration of the new antibiotic pollutants in the water body sample, wherein the concentration of the organic solvent in the redissolved aqueous solution is the optimal concentration. The invention also provides a method for optimizing enzyme-linked immunosorbent assay.

The invention relates to an anti-mouse IgG nano antibody and alkaline phosphatase fusion protein as well as a preparation method and application thereof. The anti-mouse IgG nano antibody and alkaline phosphatase fusion protein sequentially comprises an anti-mouse IgG nano antibody, a connecting peptide and shewanella alkaline phosphatase from an amino terminal to a carboxyl terminal. The fusion expression of the anti-mouse IgG nano antibody and shewanella alkaline phosphatase in escherichia coli is designed, so that various defects of a chemical labeling method can be avoided, a high-purity and high-specific activity enzyme-labeled antibody can be prepared, the preparation period of a second antibody can be shortened, the preparation process of the second antibody can be simplified, the stability of batches can be improved, and the dependence on experimental animals can be reduced. The prepared anti-mouse IgG nano antibody and alkaline phosphatase fusion protein can be applied to the field of immunodetection, such as a chemiluminescence immunoassay method, an enzyme-linked immunosorbent assay method, an enzymatic fluorescence immunoassay method, an immunoblotting method and the like.

The invention discloses a method for detecting mosquito salivary protein specific immune globulin E in animal serum through a capture method enzyme-linked immunosorbent assay, and belongs to the technical field of immunodetection. The monoclonal antibody specifically combined with the human or mouse IgE Fc fragment is used as the capture antibody, and the V5 tag is combined, so that the interference of IgG in a detection sample can be avoided, and the detection sensitivity breaks through the limit of an existing detection mode.

The invention discloses a yellow fever virus envelope protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. Yellow fever virus envelope protein E expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus envelope protein E are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the monoclonal antibody with high neutralizing activity and capable of completely protecting mice from yellow fever virus lethal attack is obtained by using the monoclonal antibody as a template, connecting the monoclonal antibody and the constant region gene to an expression vector, carrying out mammalian cell expression and purification, and carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse virus attack experiment, such that the monoclonal antibody has high neutralizing activity and can completely protect mice from yellow fever virus lethal attack; the monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention relates to the field of biological medicines, and discloses application of serum EPSTI1 in preparation of a product for diagnosing liver cancer and evaluating whether a liver cancer patient is resistant to a revatinib or not. The invention provides a detection means which is convenient and rapid in sampling, small in risk and relatively low in cost, the content of EPSTI1 in serum is detected through an enzyme-linked immunosorbent assay, a liver cancer patient and a non-liver cancer patient can be effectively distinguished, and a lovatinib drug-resistant liver cancer patient and a non-drug-resistant liver cancer patient can be effectively distinguished. Prognosis analysis shows that the serum EPSTI1 can be used as a marker for evaluating the prognosis state of the liver cancer patient taking the lenvatinib, and has important significance in improving the survival rate of the liver cancer patient.

A biomarker for systemic sclerosis, and a detection method therefor and the use thereof. The biomarker is an anti-PRMT5 antibody. The use of PRMT5 in the preparation of a drug or agent for detecting or treating systemic sclerosis, wherein patients with systemic sclerosis are distinguished from healthy controls and patients with other autoimmune diseases by means of detecting the anti-PRMT5 antibody level. The anti-PRMT5 antibody levels in patients with systemic sclerosis are closely correlated with the outcome of skin and pulmonary fibrosis in the patients, and the biomarker has potential value in prognosis evaluation. The detection method using enzyme-linked immunosorbent assay is simple, easy to perform, convenient for translation, and suitable for wide promotion in primary hospitals.

The invention relates to the field of biological detection, and discloses a plasma polypeptide marker COL1A2 related to tumor bone metastasis and pain and application. Aiming at the problem that early diagnosis of bone cancer pain and tumor bone metastasis is difficult, by detecting the level of circulating COL1A2 in plasma, it is found that the circulating COL1A2 is remarkably up-regulated in patients with tumor bone metastasis and bone cancer pain and is remarkably related to clinical pain score and tumor staging. A sensitive and specific COL1A2 detection method is established by adopting an enzyme-linked immunosorbent assay (ELISA) technology, and occurrence and staging of bone cancer pain and tumor bone metastasis can be identified with high prediction efficiency that AUC is 0.95 and 0.94 respectively. The method is low in detection cost, simple and convenient to operate and suitable for clinical popularization and application, and scientific basis and technical support are provided for screening of high-risk groups, early diagnosis of bone metastasis, pain staging and personalized treatment. The invention has important significance for optimizing diagnosis and treatment strategies of patients suffering from bone cancer pain and improving clinical transformation efficiency.