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In vitro detection kit for TAFI content

An in vitro detection and kit technology, applied in measuring devices, material analysis by observing the influence of chemical indicators, instruments, etc., can solve the problems of low detection limit, long detection time, expensive equipment, etc., and achieve detection The effect of low limit, easy operation and low cost

Inactive Publication Date: 2011-09-14
LIAONING MEDI BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a kind of in vitro detection kit of TAFI content, it has solved the defects such as expensive equipment that prior art exists, detection time is longer, compared with other detection kits, has simple and convenient operation, fast, low cost , diagnostic sensitivity, low detection limit and other advantages, as an auxiliary diagnosis for clinical prediction of cardiovascular and cerebrovascular diseases, it can significantly improve the detection rate and accuracy of the disease, which is conducive to the early detection and early treatment of the disease; reduce the number of people tested Unnecessary pain and medical expenses, improve the quality of life of the examinee

Method used

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  • In vitro detection kit for TAFI content
  • In vitro detection kit for TAFI content
  • In vitro detection kit for TAFI content

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Experimental program
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Effect test

preparation example Construction

[0031] 2. Preparation of coating antibody and enzyme-labeled antibody

[0032] Coating antibody and enzyme-labeled antibody are mouse anti-human monoclonal antibodies prepared against recombinant human TAFI antigen, and the mouse anti-human monoclonal antibodies include 3B1 and 4E7.

[0033] Hybridoma preparation: use purified human plasma TAFI as the immunogen, and immunize BALB / c mice according to conventional methods; take the splenocytes and fuse them with SP2 / 0 myeloma cells; screen the hybridoma cells, and positive wells are limited to 3 times Cloning by dilution method, and finally obtained two hybridoma cell lines that continuously and stably secrete anti-human TAFI monoclonal antibody, named 3B1 and 4E7 respectively;

[0034] Antibody identification: According to the instructions of Roche’s antibody subclass kit, the types of the two monoclonal antibodies were determined to be IgG1, κ; using simulated plasma as a control, Western blot and ELISA analysis showed that th...

Embodiment 1

[0063] Now taking human plasma as an example, the TAFI content determination is carried out on 58 patients' plasma samples diagnosed with myocardial infarction and 30 healthy plasma samples with the in vitro detection kit of TAFI content of the present invention. The specific operation steps are as follows:

[0064] Step 1 Collect the anticoagulated blood and immediately centrifuge it at 3000 rpm for 10 minutes at 4°C within 30 minutes, take the supernatant as the sample to be tested, and store it at -20°C.

[0065] Step 2 Select an appropriate amount of content in the kit to determine the TAFI content of the sample to be tested.

[0066] In step 3, use the sample diluent to dilute the sample to be tested 1600 times.

[0067] Step 4 Take 1ml of the sample diluent to dissolve the TAFI freeze-dried standard, and its concentration is 48ng / ml. Use the sample diluent to make a 2-fold gradient dilution of the 48ng / ml standard solution to make standard work with concentrations of 2...

Embodiment 2

[0079] The kit of the present invention can also measure the TAFI content in human urine. Collect human morning urine, centrifuge at 1500 rpm for 5 minutes at 4°C within 30 minutes, and take the supernatant as the sample to be tested. Use the sample diluent in the kit to dilute the sample to be tested 100 times. Subsequent steps are the same as in Embodiment 1.

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Abstract

The invention discloses an in vitro detection kit for thrombin-activatable fibrinolysis inhibitor (TAFI) content. The kit is used for the in vitro detection of the TAFI content by using the biological property that a polyamidoamine (PAMAM) dendritic polymer can be combined with an antibody and combining the traditional enzyme-linked immuno sorbent assay (ELISA) method, and the kit develops a new in vitro diagnostics method for the TAFI and improves the sensitivity of the TAFI in vitro detection. The kit comprises an antibody coated ELISA plate, a sample diluent, a TAFI standard, a detergent, a PAMAM-labeled horse radish peroxidase-linked antibody, an antibody diluent, color development buffer solution, 30 percent hydrogen peroxide, o-phenylenediamine and stop solution. The kit overcomes the defects of expensive equipment, long detection time and the like in the prior art; compared with other detection kits, the kit is simple, convenient and fast in operation, low in cost, sensitive in diagnosis and has low detection limit and the like; the kit for auxiliary diagnosis improves the detection rate and the accuracy of related diseases, and fulfills the purposes of early discovery and early treatment; and the kit reduces unnecessary pain and medicinal expense of examined persons, and improves the quality of life of the examined persons.

Description

technical field [0001] The invention relates to a kit for detecting the content of thrombin-activatable fibrinolysis inhibitor (TAFI) for clinical prediction of the possibility of occurrence of cardiovascular and cerebrovascular diseases, especially for detecting human individual biological In vitro detection kits for TAFI content in samples such as blood, serum, plasma, urine, mucus, feces, cerebrospinal fluid, pleural effusion, ascites, saliva, tissues or cells, etc. In medical testing, it mainly uses the in vitro determination of TAFI content in human individual biological samples to assist in the diagnosis and treatment of related diseases such as cardiovascular and cerebrovascular diseases, acute promyelocytic leukemia, endogenous coagulation system defects, and inflammation. Large-scale clinical census. Background technique [0002] As we all know, thrombotic disease is a kind of disease that seriously affects health, especially cardiovascular and cerebrovascular embo...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/543G01N33/577G01N21/78
Inventor 李文欣
Owner LIAONING MEDI BIOTECH CO LTD
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