Urine FXYD detection kit

A kit and quantitative detection technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of precise measurement limitations, protein interference, etc., and achieve convenient sample source, simplicity and automation, and accurate results. Effect

Inactive Publication Date: 2017-05-17
SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large number of proteins contained in serum, there are many proteins with similar amino acid sequences and higher-level structures, and these similar proteins are likely to interfere with each other during content determination, which to a certain extent leads to limitations in the accurate determination of serum FXYD sex

Method used

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  • Urine FXYD detection kit
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Preparation of enzyme-labeled anti-FXYD polyclonal antibody

[0068] The enzyme-labeled anti-FXYD polyclonal antibody is an HRP-labeled antibody, which was prepared using an activated HRP labeling kit (Jinan Taitianhe Biotechnology Co., Ltd.), and the method is as follows:

[0069] Take 100 μL of 1 mg / mL anti-FXYD polyclonal antibody diluted in 0.01M PBS buffer (pH7.4), add an equal volume of 100 μL of activated HRP with a concentration of 10 mg / mL, mix well, and adjust the pH to 9.5 with REAGENT II (approximately 10 μL), incubate in a 37°C incubator for 0.5-1h.

[0070] Use a 100 μL pipette tip to stick a small amount of sodium borohydride powder, add it to the above reaction solution, and incubate in a constant temperature incubator at 37°C for 0.5-1h.

[0071] Terminate the reaction with REAGENT III, add about three times the volume of REAGENT II (about 30 μL) to the above reaction solution, check the pH to be about 7.2, and incubate in a constant temperat...

Embodiment 2

[0073] Example 2 Anti-FXYD Antibody Coupling Magnetic Particles

[0074] 1. Preparation of reagents:

[0075] Reaction buffer: 0.05M MES, 0.5M NaCl, pH5.5;

[0076] Coupling solution: 0.15M sodium phosphate, 0.15M NaCl, pH7.5;

[0077] Blocking solution: 20mg / mL BSA+20mg / mL glycine+coupling solution;

[0078] Stock solution: 20mg / mL BSA + 20mg / mL glycine + coupling solution + 0.05% Proclin 300.

[0079] 2. Magnetic bead-conjugated antibody

[0080] Washing: take 300 μL (1 mg / mL) of magnetic beads, add 1 mL of reaction buffer and shake and wash three times, 1 min each time.

[0081] Activation: After washing the magnetic beads, remove the reaction buffer, then add the activator 200 μL EDC (1 mg / mL) and 200 μL NHS (1 mg / mL), ready for use, and shake for 60 minutes.

[0082] Coating (protein coupling): After the activation is completed, absorb 1 mL of coupling solution and shake to wash, wash three times, 1 min each time. After washing, 200 μL of coupling solution was added...

Embodiment 3

[0085] Embodiment 3 kit composition

[0086] The kit includes: magnetic particle suspension coupled with anti-FXYD antibody, FXYD quality control substance, FXYD standard substance, enzyme-labeled anti-FXYD polyclonal antibody, chemiluminescence substrate A, chemiluminescence substrate B and washing solution.

[0087] 1. Kit components

[0088] 1. The concentration of FXYD standard substance is 0, 6.25ng / mL, 18.75ng / mL, 37ng / mL, 111ng / mL, 333ng / mL, 1000ng / mL, 0.6mL each.

[0089] 2. FXYD quality control substance 200ng / mL, 0.6mL.

[0090] 3. 3 mL of magnetic particle suspension coupled with anti-FXYD antibody.

[0091] 4. Enzyme-labeled anti-FXYD polyclonal antibody 20mL.

[0092] 5. 25 mL of 25 times lotion

[0093] 6. Luminescence substrate solutions A and B are 8 mL each.

[0094] 2. Kit reagent preparation

[0095] The standard product uses Tris buffer solution of pH 7.35-7.45 as the matrix of the calibrator, calibrates high-concentration FXYD protein, and prepares a...

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Abstract

The invention discloses a urine FXYD detection kit. The kit comprises the following components: magnetic particles which are coupled with anti-FXYD antibodies, anti-FXYD polyclonal antibodies labeled with enzyme; FXYD standard substances; chemiluminescent substrate liquid. The kit can be used for quantitative determination of fibrinogen degradation product components FXYD in urine, and the kit has the characteristics of short detection time, high sensitivity, good stability, and small variation; and the kit can be used for early stage diagnosis of intestinal cancer, curative effect evaluation in treatment process, and recurrence and metastasis monitoring after treatment.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a kit for quantitatively detecting the component FXYD of the degradation product of fibrinogen, especially a kit for detecting the component FXYD of the degradation product of fibrinogen in urine. Background technique [0002] Colon cancer and rectal cancer are common malignant tumors, most of them are advanced, and their incidence rate ranks fourth among malignant tumors. With the development of the economy, people's lifestyle, living environment and dietary structure have gradually changed, resulting in an increasing trend in the incidence of colorectal cancer. Although the diagnostic level of colorectal cancer and rectal cancer has improved in recent years, the detection rate of early colorectal cancer is still low and the mortality rate is high. According to statistics, the 5-year survival rate of early colon cancer after radical surgery is about 80%, while th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/543
CPCG01N33/533G01N33/54326
Inventor 高梅娟何林富王义孙玉龙
Owner SHANGHAI LIANGRUN BIOMEDICINE TECH CO LTD
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