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Kit for detecting fibrin (ogen) degradation products (FDP) (by using emulsion immunoturbidimetry)

A latex immunization and kit technology, applied in the biological field, can solve problems such as low sensitivity, and achieve the effects of good repeatability, simple operation and high sensitivity

Active Publication Date: 2010-09-15
SHANGHAI SUNBIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a method for quantitatively detecting FDP content in samples such as plasma, serum, and urine in view of the low sensitivity of the existing latex immunoturbidimetric method for detecting FDP, and the operation is simple, fast, quantitatively accurate and sensitive. Higher diagnostic kits

Method used

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  • Kit for detecting fibrin (ogen) degradation products (FDP) (by using emulsion immunoturbidimetry)
  • Kit for detecting fibrin (ogen) degradation products (FDP) (by using emulsion immunoturbidimetry)
  • Kit for detecting fibrin (ogen) degradation products (FDP) (by using emulsion immunoturbidimetry)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: composition and preparation method of the kit of the present invention

[0048] Reagent R2 is the preparation of anti-human FDP antibody latex reagent: reconstitute the lyophilized product containing 500 μg of mouse anti-human FDP monoclonal antibody (purchased from American Diagnostica Inc., product number ADINo.313) with 0.5 ml of deionized distilled water, and take 50 μl of solution was diluted with 3ml of 0.1M sodium chloride solution (containing 0.1% sodium azide (g / ml)), and 2.0ml of 0.05M glycine buffer solution (pH2.3, 0.1M sodium chloride) was added to the solution. The pH value is 2.8. The acidified antibody solution was kept at room temperature (20-25° C.) for 30-40 min, and then 100 μl of 1 M tris-hydroxymethylaminomethane solution was added to the solution to adjust the pH value to 7.5-8.0.

[0049] Get 1.25ml 4% (g / ml) polyvinylbenzyl chloride latex particle suspension (USA INVITROGEN company, product number is C37345, average particle diame...

Embodiment 2

[0055] Embodiment 2: the assay method of kit of the present invention

[0056] Add 0.5ml of distilled water to dissolve the FDP calibrator in the kit, and prepare 6 calibrator solutions with different concentrations with the diluent, the concentrations are 0.958, 1.915, 3.830, 7.660, 15.320, and 30.640 μg / ml. Take the operation of CA550 hemagglutination analyzer in East Asia, Japan as an example: the reaction temperature is 37°C, the measurement wavelength is 575nm, take 10 μl of calibrator solution with different concentrations, add 10 μl of diluent, react for 30 seconds, add reagent R1100 μl, and react for 60 seconds Add the reagent R2100 μ l that embodiment 1 prepares afterward, measure the absorbance value (A 1 、A 2 ), calculate the absorbance difference ΔA=A 2 -A 1 , repeat the measurement 3 times for each tube, take the average of the absorbance difference ΔA measured three times for each calibration tube as the ordinate, and the corresponding concentration as the abs...

Embodiment 3

[0059] Embodiment 3: Analytical performance evaluation of the kit of the present invention

[0060] 1. Linear range

[0061] The high-concentration sample (30.42 μg / ml) of FDP close to the upper limit of the linear range was diluted with FDP diluent by 1 / 2, 1 / 10, 1 / 15, 1 / 30, and 1 / 60, and a total of 6 Each concentration is detected with the method described in Example 2, and each concentration is repeatedly measured 3 times, and the mean value of the measured concentration and the theoretical concentration are carried out to linear regression analysis, and the calculation regression equation is Y=1.03917X-0.08011, the correlation The coefficient is r=0.9996, showing that the correlation of the kit of the present invention is good in the linear range of 0.50 μg / ml~30.0 μg / ml, see figure 2 .

[0062] 2. Sensitivity is the lowest detection limit

[0063]Take 5% human serum albumin as a blank sample, measure according to the method described in Example 2, repeat the measuremen...

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Abstract

The invention relates to the field of biotechnology and discloses a kit for detecting fibrin (ogen) degradation products (FDP) by using emulsion immunoturbidimetry. The kit comprises an anti-human FDP antibody emulsion which is formed by performing chemical crosslinking of acidulated anti-human FDP antibodies and polyvinyl benzyl chloride emulsion particles and sealing the reaction product. The kit of the invention has the advantages that: the emulsion immunoturbidimetry is adopted to detect the FDP content, so the sensitivity is high and can reach 0.10 mu g / ml; the stability is high, the operation is simple and quick and it only takes 5 to 10 minutes to obtain a detection result; the specificity is high and the detection avoids interference effectively; and the quantification is accurateand the application range is wide.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting the content of fibrin (ogen) degradation products (FDP) by latex immunoturbidimetry. Background technique [0002] The fibrinolytic system, referred to as the fibrinolytic system, is the most important anticoagulant system in the human body. It plays an important role in maintaining the normal permeability of the blood vessel wall, maintaining the blood flow state and tissue repair. It consists of 4 main parts: Lysinogen, plasminogen activators (eg, t-PA, u-PA), plasmin, plasmin inhibitors. Fibrin (ogen) Degradation Products (Fibrin (ogen) Degradation Products, FDP) is the action of plasmin produced after the fibrinolytic system is activated, the fibrinogen and fibrin in the human body are degraded, and the various products produced A general term for dimers, multimers and complexes, and the generated FDP reflects the total level of fibrinolytic activity in the b...

Claims

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Application Information

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IPC IPC(8): G01N33/86G01N33/531
Inventor 谢永华朱美萍许付
Owner SHANGHAI SUNBIO TECH
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