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Competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and preparation method thereof

A detection kit and latex particle technology, applied in the biological field, can solve the problems of high difficulty and long detection time of BNP detection by sandwich method, and achieve the effects of low detection cost, simple detection method and good specificity.

Inactive Publication Date: 2012-09-19
王钊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the current sandwich method is difficult to detect BNP and the detection time is long, and to provide a fast and easy-to-detect competition method latex particle-enhanced immune turbidimetric BNP detection kit and its preparation method

Method used

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  • Competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and preparation method thereof
  • Competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The invention consists of latex particles and reaction buffer.

[0031] The latex particles are coated with the following artificially synthesized amino acid sequence: FGRKMDR-X; X consists of 2-4 K. The reaction buffer contains monoclonal antibody against FGRKMDR epitope. The FGRKMDR is an existing amino acid sequence.

[0032] The artificially synthesized amino acid sequence: FGRKMDR-X, was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. The monoclonal antibody against FGRKMDR epitope was provided by Abcom.

[0033] (a) The preparation method of the latex particles consists of the following steps:

[0034] (a1) Use EDC and S-NHS to activate the latex particles, the activation time is 15min, the activated substance is centrifuged at 22000rpm for 10min, the supernatant is removed, and the HEPES buffer is used to redissolve;

[0035] (a2) adding the artificially synthesized amino acid sequence FGRKMDRKK and reacting at room temperature for 2 hours;

[0036...

Embodiment 2

[0047]The difference between this example and Example 1 is that the artificially synthesized amino acid sequences coated with latex particles are different; meanwhile, the final concentration of the monoclonal antibody against FGRKMDR epitope used in this example is 0.05 mg / ml. The artificially synthesized amino acid sequence used in this example is: FGRKMDRKKK. The test results are shown in Table 1.

Embodiment 3

[0049] The difference between this example and Example 1 is that the artificially synthesized amino acid sequences coated with latex particles are different; meanwhile, the final concentration of the monoclonal antibody against FGRKMDR epitope used in this example is 0.1 mg / ml. The artificially synthesized amino acid sequence used in this example is: FGRKMDRKKKK. The test results are shown in Table 1.

[0050] Table 1

[0051] sample Example 1 Example 2 Example 3 Controlled experiment 1 133.6 141.3 137.5 128.3 2 25.2 27.3 26.4 22.3 3 400.1 440.2 439.7 419.5 4 175.3 179.8 180.2 170.0 5 234.5 242.4 236.3 253.3 6 65.5 68.2 66.7 73.2 7 85.3 88.6 88.2 82.3 8 12.5 13.1 12.7 14.4 9 542.7 553.3 547.5 490.6

[0052] Through the above table 1, it can be effectively shown that the content of BNP (pg / ml) in the sample can be effectively detected by the present invention, and its accuracy is high...

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Abstract

The invention discloses a competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and a preparation method thereof, and solves the problems of high difficulty in BNP assay, long time for assay and the like in the current sandwich method. The kit disclosed by the invention is composed of latex particles and reaction buffer, and is characterized in that the latex particles are coated with the following artificially-synthesized amino acid sequences: FGRKMDR-X, and X consists of 2 to 4 Ks. The invention further provides a preparation method of the kit.. The kit disclosed by the invention has the advantages of high accuracy, short time for assay, high reliability, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a competition method latex particle enhanced immune turbidimetric BNP detection kit, and the invention also relates to a preparation method of the kit. Background technique [0002] B-type natriuretic peptide is also called brain natriuretic peptide (BNP). As a quantitative marker of heart failure, BNP not only reflects left ventricular systolic dysfunction, but also reflects left ventricular diastolic dysfunction, valve dysfunction and right ventricular dysfunction. Taking BNP 100pg / ml as the cut-off value has a negative predictive value of 90%, which can reduce clinical uncertainty by 74%; while BNP exceeding 400pg / ml indicates that the possibility of heart failure in patients reaches 95%. While BNP at 100-400pg / ml may be caused by lung disease, right heart failure, pulmonary embolism and other conditions. BNP levels at emergency department visits and post-treatment changes in pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531G01N21/31
Inventor 王钊
Owner 王钊
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