CN111693700A - Use of factor B antibodies in preparation of detection kit - Google Patents
Use of factor B antibodies in preparation of detection kit Download PDFInfo
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- CN111693700A CN111693700A CN202010647899.1A CN202010647899A CN111693700A CN 111693700 A CN111693700 A CN 111693700A CN 202010647899 A CN202010647899 A CN 202010647899A CN 111693700 A CN111693700 A CN 111693700A
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Abstract
The invention provides an application of a factor B antibody in preparing a kit for detecting peripheral serum, whole blood or peripheral blood for diagnosing heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, tumor, bacterial infection, viral infection or tumor. The kit disclosed by the invention coats the factor B monoclonal antibody or the factor B monoclonal antibody and the polyclonal antibody, and the colloidal gold is combined with the polyclonal antibody, so that the sensitivity of the kit can be improved, and the kit can be used together with the polyclonal antibody and applied to immunoturbidimetry (immunotransmission turbidimetry, immunoscattering turbidimetry, immunolatex turbidimetry), ELISA (enzyme-linked immunosorbent assay), chemiluminescence, microfluidic chip method, colloidal gold percolation method and chromatography. The invention also provides a kit, which adopts a method of combining B factor antigen-antibody (comprising monoclonal antibody and polyclonal antibody) for detection and can be applied to diagnosis of heart or cerebral thrombus heart or cerebral infarction cerebral hemorrhage heart or cerebral embolism, bacterial infection or virus infection and tumor.
Description
Technical Field
The invention belongs to the field of bioengineering, and relates to a detection kit, in particular to an application of a factor B in preparing a kit for detecting peripheral serum, whole blood or peripheral blood for diagnosing heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, bacterial infection, viral infection and tumor.
Background
Factor B is a single-chain protein encoded by a gene located on human chromosome 6 and comprising five distinct domains, has a molecular weight of about 93kD, is mainly secreted by liver cells, and can be produced by other cells such as monocytes, macrophages, fibroblasts, dendritic cells, small intestine and renal tubules, and the like, and is physiologically present at a concentration of about 200 μ g/ml in plasma.
Factor B is an important protein of the alternative complement pathway, which is cleaved by factor D into two fragments, Ba (33kD) and Bb (60 kD). The Ba region contains 3 Complement Control Protein (CCP) domains, and the Bb region contains a von Willebrand factor type a domain (VWA) and a S1 peptidase domain. The CCP domain of Ba and the VWA domain of Bb play a very important role in the binding of factor B to C3B. The S1 peptidase domain contains important catalytically active components bypassing the C3 convertase and the C5 convertase.
The B factor kit is originally used for diagnosing diseases caused by peripheral blood B factor reduction, is occasionally found to be relevant to diagnosis of heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage and heart or cerebral embolism in screening of hundreds of antibodies, and is also relevant to diagnosis indexes of gram-positive bacteria, gram-negative bacteria, virus infection and tumor diagnosis such as thyroid cancer, liver cancer, gastric cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophagus cancer, nasopharyngeal carcinoma, prostate cancer, colorectal cancer, kidney cancer, pancreatic cancer and bladder cancer.
Disclosure of Invention
The invention aims to provide the application of a factor B antibody, and the application of the factor B antibody aims to solve the technical problems that the means for detecting the diagnosis of heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, bacterial infection, thyroid cancer, liver cancer, gastric cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostatic cancer, colon cancer, rectal cancer, renal cancer, pancreatic cancer and bladder cancer or viral infection is limited and the time is long in the prior art.
The invention provides an application of a factor B antibody in preparing a kit for detecting peripheral serum, whole blood or peripheral blood for diagnosing heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, bacterial infection or viral infection or tumor.
Further, the tumor is thyroid cancer, liver cancer, stomach cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostatic cancer, colon cancer, rectal cancer, kidney cancer, pancreatic cancer or bladder cancer.
Further, the kit comprises a detection plate, a rabbit anti-B factor antibody, a goat anti-B factor antibody conjugate marked by colloidal gold, a confining liquid, a washing liquid, a positive reference product and a negative reference product, wherein the detection plate consists of a plastic box bottom box, a water absorption layer is arranged in the plastic box bottom box, a nitrocellulose membrane is arranged on the water absorption layer, a filter screen is covered on the nitrocellulose membrane, a plastic box surface cover is arranged on the filter screen, a detection reaction hole is arranged in the center of the plastic box surface cover, the conjugate of the rabbit anti-B factor antibody and the goat anti-B factor antibody marked by colloidal gold is dotted on the nitrocellulose membrane corresponding to the detection reaction hole, the confining liquid is a phosphate solution containing albumin, the pH value of the bovine serum phosphate solution is 7.0-7.5, and the weight percentage of the bovine serum albumin in the phosphate solution is 1.0%, the washing solution is a Tris-HCl solution containing Tween-20, the pH of the Tris-HCl solution is 7.0-7.5, the weight percentage of the Tween-20 in the Tris-HCl solution is 0.05%, the mass of the goat anti-factor B antibody in each milliliter of colloidal gold conjugate is 0.025 mg, and the positive reference substance is a factor B antigen with the concentration of more than 0.25 g/L.
Furthermore, the rabbit anti-factor B antibody is a monoclonal antibody or a polyclonal antibody.
Further, colloidal gold filtration, chromatography, chemiluminescence, and detection were performed.
Furthermore, ELISA method, immunoturbidimetry, immunotransmission turbidimetry, immunoscattering turbidimetry or immunolatex enhanced turbidimetry is adopted for carrying out clinical examination and routine detection.
Furthermore, a micro-fluidic chip is adopted for detection.
Furthermore, the rabbit anti-factor B antibody and the monoclonal antibody or the polyclonal antibody.
The invention also provides a kit, which adopts a method of combining the B factor antigen and the B factor antibody for detection.
The invention uses the combination of the B factor antibody and the antigen for detecting the early diagnosis of peripheral serum, whole blood or peripheral blood for diagnosing heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, bacterial infection or virus infection and tumor, thyroid cancer, liver cancer, gastric cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostatic cancer, colorectal cancer, renal cancer, pancreatic cancer and bladder cancer.
Furthermore, the rabbit anti-factor B antibody is a monoclonal antibody or a polyclonal antibody, so that the sensitivity and the accuracy of the reagent are improved (comprising the monoclonal antibody or the polyclonal antibody of a mouse, a rabbit, a sheep or a horse).
The detection principle of the invention is as follows: binding reaction of B factor antibody and B factor antigen in serum.
Compared with the prior art, the invention has remarkable technical progress. The kit can be used for detecting the B factor antigen in a human serum sample, and can be used for diagnosing heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, bacterial infection or virus infection and tumors such as thyroid cancer, liver cancer, gastric cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostatic cancer, colorectal cancer, renal cancer, pancreatic cancer and bladder cancer. By adopting the kit, a detection report can be received within 1-5 minutes, and the detection result is accurate and convenient.
Detailed Description
Example 1 preparation of a kit
1.1 preparation process of colloidal gold conjugate;
1.1.1 preparing colloidal gold;
1.1.1.10.05% gold chloride solution 300 ml, heating to boil;
1.1.1.2 add 3.6 ml of 5.0% trisodium citrate;
1.1.1.3, continuously heating and boiling for 15 minutes, stopping heating, cooling to room temperature, and recovering the volume to the original volume by using distilled water;
1.1.2 labeling with colloidal gold;
1.1.2.1 colloidal gold 100 mL;
1.1.2.2 adding 2.4 ml of 1.0% anhydrous sodium carbonate solution, and adjusting the pH value to 6.4-7.2;
1.1.2.3 adding 2.5 mL goat anti-factor B antibody (1.0mg/mL), standing at room temperature for 30 min;
1.1.2.4 stirring and adding 10XTris-HCl20 ml to regulate pH to 7.2-7.5;
1.1.2.5 stirring and adding 2 g of BSA, 1 g of PEG20000 and 0.2 ml of ProClin-300;
1.2 detection board preparation process;
1.2.1 assembling a detection plate, taking a plastic box bottom cover, placing a water absorption layer, placing a nitrocellulose membrane on the water absorption layer, covering a plastic box surface cover, and compressing;
1.2.2 detection of Point antibodies: rabbit anti-factor B antibody (1.0mg/mL)1.0 mL 1.0M NaHCO was added30.1 ml, and mixing evenly;
1.2.3 taking 2.0uL of detection point antibody, and adding the detection point antibody on a nitrocellulose membrane at the center of a reaction hole of the detection plate to prepare a detection plate;
1.3 preparing sealing liquid and washing liquid;
1.3.1 preparing a sealing liquid, 1.0 percent of bovine serum albumin and 0.02M phosphate solution with the pH value of 7.0-7.5;
1.3.2 preparing a washing solution, 0.05 percent of Tween-20 and 0.05M of Tris-HCl solution with the pH7.0-7.5;
1.4 determining a normal reference range, based on the normal reference range of the B factor antigen determination kit, wherein the content of the B factor antigen is 50mg/L-250mg/L, and because the method is qualitative detection and the content of the B factor antigen of a sample does not develop color at a detection point in the normal reference range, selecting a sample with the content of the B factor antigen of about 250mg/L as sensitivity detection;
1.4.1 methods of experiment:
1. before use, the reagent and the sample are taken out, and are placed for 15-30 minutes at room temperature to be recovered to the room temperature;
2. taking out the detection plate;
3. adding 2 drops of sealing liquid into the center of a reaction hole of the detection plate until the sealing liquid completely permeates;
4. adding 40 microliters of the sample to be detected into the center of the reaction hole of the detection plate by using a pipettor until the sample completely permeates into the reaction hole;
5. adding 4 drops of washing liquid into the center of the reaction hole of the detection plate until the washing liquid completely permeates into the reaction hole;
6. adding 3 drops of colloidal gold conjugate in the center of the reaction hole of the detection plate until the colloidal gold conjugate completely permeates;
7. adding 4 drops of washing liquid into the center of the reaction hole of the detection plate, and observing the result visually after the washing liquid completely permeates;
[ interpretation of test results ]
When a user performs sample detection, the following situations may occur:
a. a red spot is arranged at the center detection point of the reaction hole of the detection plate, and a positive result is obtained;
b. the detection point at the center of the reaction hole of the detection plate has no red spot, and the result is negative.
[ REFERENCE VALUES (REFERENCE RANGE) ]
Based on the normal reference range of the B factor antigen determination kit, the content of the B factor antigen is 50mg/L-250mg/L, no red spot exists at the center detection point of the reaction hole of the detection plate in the normal reference range of the sample B factor antigen content, and a negative result is displayed;
normal reference range: negative results are shown (indicating that the antigen content of the factor B is between 50mg/L and 250 mg/L).
[ interpretation of test results ]
1. Negative result, the content of B factor antigen in the sample is between 50mg/L and 250mg/L and is in normal range;
2. a positive result indicates that the sample has an abnormal B factor antigen content of > 250 mg/L.
[ limitations of assay methods ]
1. The method can only qualitatively determine the B factor in the human serum sample and is only used as auxiliary diagnosis;
2. stability of reaction spots: within 30 minutes the positive spots did not fade and the negative results remained unchanged.
20 examples of detection results: (patients with cerebral thrombosis, cerebral infarction, cerebral hemorrhage, and cerebral embolism)
| Positive for | Positive for | Positive for | Positive for | Positive for |
| Positive for | Positive for | Positive for | Positive for | Positive for |
| Positive for | Positive for | Positive for | Positive for | Positive for |
| Positive for | Positive for | Positive for | Positive for | Positive for |
In 20 healthy persons (all negative in liver function, kidney function, HBsAg, HIV, hepatitis C, etc.), the negative is 50mg/L-250 mg/L.
| Negative of | Negative of | Negative of | Negative of | Negative of |
| Negative of | Negative of | Negative of | Negative of | Negative of |
| Negative of | Negative of | Negative of | Negative of | Negative of |
| Negative of | Negative of | Negative of | Negative of | Negative of |
Results of 4 patients of 20 cases of heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage and heart or cerebral embolism are compared with results of 20 healthy patients, and the clinical diagnosis is completely met.
Results from 60 of 10 tumor patients: (thyroid cancer, liver cancer, stomach cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostate cancer, colorectal cancer, kidney cancer, pancreatic cancer, bladder cancer)
Example 2: immune latex turbidimetry kit
The instrument comprises the following steps: 7100 Biochemical instrument
The operation method comprises the following steps: the samples were assayed using Hitachi 7100 Biochemical Analyzer. In the step of measuring the sample by using a biochemical analyzer, after the sample is added with the reagent I R1, the sample is incubated at 37 ℃ for 5min, the sample A1 at the 1 st point is read, the reagent II R2 is added, the sample is incubated at 37 ℃ for 5min, the sample A2 at the 2 nd point is read, and the sample A is sample A2-sample A1; the method also comprises a step of measuring the calibrator by using a Hitachi 7100 biochemical analyzer, wherein in the step of measuring the calibrator by using the biochemical analyzer, the calibrator is added into a reagent I R1, then the mixture is incubated at 37 ℃ for 5min, the calibrator A1 at the 1 st point is read, the reagent II R2 is added, the mixture is incubated at 37 ℃ for 5min, and the calibrator A2 at the 2 nd point is read, wherein the calibrator A is calibrator A2-calibrator A1;
the parameters measured above were: temperature 37 ℃, wavelength 600nm, R1: 160uL R2: 40 uL. Sample or calibrator 3 μ l, R1: 240 μ l, R2: 60 μ l, reaction time 10 min.
20 normal persons (negative for liver function, kidney function, HBsAg, HIV, hepatitis C, etc.)
| 156mg/L | 216mg/L | 221mg/L | 165mg/L | 228mg/L |
| 173mg/L | 166mg/L | 190mg/L | 170mg/L | 209mg/L |
| 184mg/L | 175mg/L | 185mg/L | 169mg/L | 189mg/L |
| 207mg/L | 194mg/L | 176mg/L | 178mg/L | 174mg/L |
Concentrations between 50mg/L and 250mg/L are normal.
The clinical diagnosis results of 20 healthy people are between 50mg/L and 250mg/L, are negative and meet the clinical diagnosis standard.
60 cases of tumor patients (thyroid cancer, liver cancer, stomach cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostate cancer, colorectal cancer, kidney cancer, pancreatic cancer, bladder cancer)
Concentrations between 50mg/L and 250mg/L are normal.
The results of 60 cases of clinical diagnosis are below 50mg/L or above 250mg/L, are positive and accord with the clinical diagnosis standard.
Example 3: 20 samples of cerebral hemorrhage (immunoturbidimetry)
The instrument comprises the following steps: 7100 Biochemical instrument
The operation method is the same as that of example 2
Kit parameters: wavelength 600nm, R1: 160uL R2: 40 uL.
20 examples of detection results:
concentrations between 50mg/L and 250mg/L are normal.
The clinical diagnosis result of 20 patients with cerebral hemorrhage is more than 250mg/L, is positive and accords with the clinical diagnosis standard.
Example 4: 20 samples of cardiac or cerebral infarction (immunoturbidimetry)
The instrument comprises the following steps: 7100 Biochemical instrument
The operation method is the same as that of example 2
Parameters are as follows: wavelength 340nm, R1: 160uL R2: 40 uL.
20 cases of test results
| 362mg/L | 443mg/L | 345mg/L | 312mg/L | 346mg/L |
| 476mg/L | 479mg/L | 264mg/L | 407mg/L | 371mg/L |
| 415mg/L | 502mg/L | 294mg/L | 285mg/L | 362mg/L |
| 360mg/L | 304mg/L | 305mg/L | 284mg/L | 296mg/L |
Concentrations between 50mg/L and 250mg/L are normal.
The clinical diagnosis result of 20 patients with cardiac or cerebral infarction is more than 250mg/L, is positive and meets the clinical diagnosis standard.
Example 5: 20 samples of cardiac or cerebral thrombosis (immunoturbidimetry)
The instrument comprises the following steps: 7100 Biochemical instrument
The operation method is the same as that of example 2
Parameters are as follows: wavelength 600nm, R1: 160uL R2: 40 uL.
20 cases of test results
| 399mg/L | 450mg/L | 392mg/L | 344mg/L | 426mg/L |
| 284mg/L | 436mg/L | 388mg/L | 470mg/L | 491mg/L |
| 321mg/L | 402mg/L | 280mg/L | 493mg/L | 325mg/L |
| 347mg/L | 283mg/L | 301mg/L | 369mg/L | 293mg/L |
Concentrations between 50mg/L and 250mg/L are normal.
The clinical diagnosis result of 20 patients with cardiac or cerebral thrombosis is more than 250mg/L, is positive and accords with the clinical diagnosis standard.
Example 6: 20 samples of cardiac or cerebral embolism (immunoturbidimetry)
The instrument comprises the following steps: 7100 Biochemical instrument
The operation method is the same as that of example 2
Parameters are as follows: wavelength 600nm, R1: 160uL R2: 40 uL.
20 examples of detection results:
concentrations between 50mg/L and 250mg/L are normal.
The clinical diagnosis result of 20 patients with cardiac or cerebral embolism is more than 250mg/L, is positive and meets the clinical diagnosis standard.
Example 7: 20 examples chemiluminescence method
The chemiluminescence detection method comprises the following steps:
1. the instrument comprises the following steps: MPC-1 chemiluminescence determinator
2. Fixing the anti-human factor B antibody on the surface of a solid phase, adding a sample to be detected and a marked anti-human factor B antibody, incubating, and then washing with a washing solution; meanwhile, setting a reference of a factor B standard substance;
3. adding a chemiluminescent substrate working solution, and standing for a period of time;
4. and measuring the luminous value to obtain the B factor concentration value of the sample to be detected.
20 examples of detection results: 20 patients with cardiac or cerebral infarction
| 386mg/L | 350mg/L | 319mg/L | 495mg/L | 255mg/L |
| 263mg/L | 374mg/L | 405mg/L | 353mg/L | 368mg/L |
| 415mg/L | 284mg/L | 271mg/L | 340mg/L | 373mg/L |
| 352mg/L | 316mg/L | 265mg/L | 344mg/L | 346mg/L |
Concentrations between 50mg/L and 250mg/L are normal.
The results of 20 cases of clinical diagnosis are more than 250mg/L, are positive and accord with the clinical diagnosis standard.
Example 8: 50 samples of gram-positive bacteria, gram-negative bacteria and virus infection (chemiluminescence method) 50 test results
Concentrations between 50mg/L and 250mg/L are normal.
The clinical diagnosis result of 50 cases of patients infected by gram-positive bacteria, gram-negative bacteria and viruses is more than 250mg/L, is positive and accords with the clinical diagnosis standard.
The above examples of the present invention are merely examples for clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (9)
- Use of a factor B antibody in the preparation of a kit for detecting peripheral serum, whole blood or peripheral blood for diagnosing heart or cerebral thrombosis, heart or cerebral infarction, cerebral hemorrhage, heart or cerebral embolism, bacterial infection or viral infection or tumor.
- 2. Use according to claim 1, characterized in that: the tumor is thyroid cancer, liver cancer, gastric cancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharyngeal cancer, prostatic cancer, colon cancer, rectal cancer, renal cancer, pancreatic cancer or bladder cancer.
- 3. Use according to claim 1, characterized in that: the kit comprises a detection plate, a rabbit anti-B factor antibody, a goat anti-B factor antibody conjugate marked by colloidal gold, a sealing liquid, a washing liquid, a positive reference product and a negative reference product, wherein the detection plate consists of a plastic box bottom box, a water absorption layer is arranged in the plastic box bottom box, a nitrocellulose membrane is arranged on the water absorption layer, a filter screen is covered on the nitrocellulose membrane, a plastic box surface cover is arranged on the filter screen, a detection reaction hole is arranged in the center of the plastic box surface cover, the conjugate of the rabbit anti-B factor antibody and the goat anti-B factor antibody marked by the colloidal gold is added on the nitrocellulose membrane corresponding to the detection reaction hole, the sealing liquid is a phosphate solution containing bovine serum albumin, the pH value of the phosphate solution is between 7.0 and 7.5, and the weight percentage of the bovine serum albumin in the phosphate solution is 1.0 percent, the washing solution is a Tris-HCl solution containing Tween-20, the pH of the Tris-HCl solution is 7.0-7.5, the weight percentage of the Tween-20 in the Tris-HCl solution is 0.05%, the mass of the goat anti-factor B antibody in each milliliter of colloidal gold conjugate is 0.025 mg, and the positive reference substance is a factor B antigen with the concentration of more than 0.25 g/L.
- 4. Use according to claim 1, characterized in that: the rabbit anti-factor B antibody is a monoclonal antibody or a polyclonal antibody.
- 5. Use according to claim 1, characterized in that: detecting by colloidal gold percolation, chromatography, and chemiluminescence.
- 6. Use according to claim 1, characterized in that: and performing clinical examination and routine detection by adopting an ELISA method, an immunoturbidimetry method, an immunotransmission turbidimetry method, an immunoscattering turbidimetry method or an immunolatex enhanced turbidimetry method.
- 7. Use according to claim 1, characterized in that: and detecting by adopting a micro-fluidic chip.
- 8. Use according to claim 1, characterized in that: the rabbit anti-factor B antibody and the monoclonal antibody or the polyclonal antibody.
- 9. A kit, characterized in that: the detection is carried out by adopting a method of combining the B factor antigen and the B factor antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010647899.1A CN111693700A (en) | 2020-07-07 | 2020-07-07 | Use of factor B antibodies in preparation of detection kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010647899.1A CN111693700A (en) | 2020-07-07 | 2020-07-07 | Use of factor B antibodies in preparation of detection kit |
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| CN111693700A true CN111693700A (en) | 2020-09-22 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202010647899.1A Pending CN111693700A (en) | 2020-07-07 | 2020-07-07 | Use of factor B antibodies in preparation of detection kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114252623A (en) * | 2020-09-23 | 2022-03-29 | 上海怡珏生物科技有限公司 | Use of FN in preparing detection kit |
| CN114487410A (en) * | 2020-11-13 | 2022-05-13 | 上海怡珏生物科技有限公司 | Use of factor H antibody in the preparation of a kit for detecting hemodialysis, peritoneal dialysis, peritonitis or glioma |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050277158A1 (en) * | 2004-06-10 | 2005-12-15 | Ge Chen | Method and kit for donor specific complement-fixing antibodies crossmatch |
| CN101158689A (en) * | 2007-11-19 | 2008-04-09 | 广西师范大学 | Immunoresonance Scattering Spectroscopy for Detection of Factor B |
| CN103743912A (en) * | 2013-12-31 | 2014-04-23 | 浙江爱康生物科技有限公司 | B factor determination kit and preparation method thereof |
| CN106461661A (en) * | 2014-05-02 | 2017-02-22 | Uab研究基金会 | Methods and compositions for diagnosis and treatment of meningitis |
-
2020
- 2020-07-07 CN CN202010647899.1A patent/CN111693700A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050277158A1 (en) * | 2004-06-10 | 2005-12-15 | Ge Chen | Method and kit for donor specific complement-fixing antibodies crossmatch |
| CN101158689A (en) * | 2007-11-19 | 2008-04-09 | 广西师范大学 | Immunoresonance Scattering Spectroscopy for Detection of Factor B |
| CN103743912A (en) * | 2013-12-31 | 2014-04-23 | 浙江爱康生物科技有限公司 | B factor determination kit and preparation method thereof |
| CN106461661A (en) * | 2014-05-02 | 2017-02-22 | Uab研究基金会 | Methods and compositions for diagnosis and treatment of meningitis |
Non-Patent Citations (2)
| Title |
|---|
| 杨国灿;张芬芬;: "血清B因子与肿瘤疾病的相关关系" * |
| 蔡莉;王雨生;胡丹;王海燕;张自峰;: "视神经炎98例病因学临床分析" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114252623A (en) * | 2020-09-23 | 2022-03-29 | 上海怡珏生物科技有限公司 | Use of FN in preparing detection kit |
| CN114487410A (en) * | 2020-11-13 | 2022-05-13 | 上海怡珏生物科技有限公司 | Use of factor H antibody in the preparation of a kit for detecting hemodialysis, peritoneal dialysis, peritonitis or glioma |
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