CN110531067B

CN110531067B - Detection kit for fibrin and fibrinogen degradation products

Info

Publication number: CN110531067B
Application number: CN201910818580.8A
Authority: CN
Inventors: 史建国; 李静; 王珍; 董雯; 胡晓飞; 包兴艳; 李志明
Original assignee: Biobase Biodustry Shandong Co Ltd
Current assignee: Biobase Biodustry Shandong Co Ltd
Priority date: 2019-08-30
Filing date: 2019-08-30
Publication date: 2021-01-22
Anticipated expiration: 2039-08-30
Also published as: CN110531067A

Abstract

The invention discloses a detection kit for fibrin and fibrinogen degradation products, and relates to the technical field of clinical in vitro diagnosis. The detection kit comprises a reagent R1, a reagent R2 and a calibrator, wherein the reagent R1 comprises the following components in percentage by weight: 30-100 mmol/L of buffer solution, 1 part of surfactant: 1-5 ml/L; the components and contents of the reagent R2 are as follows: 30-100 mmol/L of buffer solution, 0.1-0.5% of antihuman FDP monoclonal antibody latex particles, 2% of surfactant: 1-5 ml/L, 0.1-0.5 g/L bovine serum albumin, 1-5 ml/L fatty acid sorbitan, 1-5 ml/L betaine and 1-10 g/L polyvinylpyrrolidone. The invention can further improve the stability on the basis of ensuring the detection accuracy, linear correlation and analysis sensitivity.

Description

Detection kit for fibrin and fibrinogen degradation products

Technical Field

The invention relates to the technical field of clinical in-vitro detection, and in particular relates to a detection kit for fibrin and fibrinogen degradation products.

Background

The fibrin degradation product is a degradation product (FDP) produced after fibrinogen and fibrin are decomposed by plasma cellulose, and mainly reflects a dissolving function of fibrin. FDP is detected simultaneously with other indexes, so that primary or secondary hyperfibrino lysis can be identified. Clinically, FDP is taken as a reference index of various thrombotic diseases and is listed as one of the conventional indexes for diagnosis of a disseminated intravascular coagulation laboratory.

The detection of plasma fibrin degradation products is to quantitatively determine the content of FDP in serum, and the detection result is expressed by milligrams of FDP (mg/L) in each liter of plasma, and the content of FDP can reflect the intensity of fibrinolytic activity in vivo. FDP can inhibit fibrin formation, has antithrombin effect, and inhibits platelet adhesion, aggregation and release.

At present, the clinical detection methods for FDP mainly comprise a latex immunoturbidimetry method, an enzyme-linked immunosorbent assay, an ethanol gel test method, a latex agglutination method and the like, wherein the latex immunoturbidimetry method for detecting FDP is an analysis method which does not need to pre-treat samples, has low technical and equipment requirements and has higher precision and specificity.

Disclosure of Invention

The invention provides a reagent kit for detecting fibrin and fibrinogen degradation products, which can further improve the stability on the basis of ensuring the detection accuracy, linear correlation and analysis sensitivity.

In order to solve the technical problems, the invention provides the following technical scheme:

the invention provides a detection kit for fibrin and fibrinogen degradation products, which comprises a reagent R1, a reagent R2 and a calibrator, wherein:

the components and contents of the reagent R1 are as follows:

buffer solution 30-100 mmol/L

11-5 ml/L of surfactant;

the components and contents of the reagent R2 are as follows:

further, the buffer solution is Tris-HCL buffer solution, and the pH value is 8.0.

Further, the betaine is cocamidopropyl betaine.

Further, the surfactant 1 and the surfactant 2 are one or more of Proclin300, Triton X-100 and polyoxyethylene lauryl ether.

Further, the anti-human FDP monoclonal antibody latex particles comprise one or more of a mouse anti-human FDP monoclonal antibody, a rabbit anti-human FDP monoclonal antibody and a sheep anti-human FDP monoclonal antibody.

Further, the components and contents of the reagent R1 are as follows:

buffer 50mmol/L

13 ml/L of surfactant;

the components and contents of the reagent R2 are as follows:

further, the volume ratio of the reagent R1 to the reagent R2 is 1: 1.

Compared with the prior art, the invention has the following beneficial effects:

the reagent kit for detecting fibrin and fibrinogen decomposition products adds a stabilizer consisting of four components of BSA, sorbitan fatty acid, betaine and polyvinylpyrrolidone into a reagent R2, wherein the BSA can play a good role in protecting and stabilizing colloid of antibodies crosslinked on the surfaces of latex particles, and the sorbitan fatty acid and the betaine are adsorbed on the surfaces of the latex particles and used for reducing the surface tension of the latex particles, so that the latex particles are not easy to aggregate, the stability of the latex particles in a solution is effectively improved, and the polyvinylpyrrolidone can improve the density and viscosity of the solution, so that the latex particles can be suspended in the solution for a long time and can not be settled to play a good role in stabilizing. The invention further improves the stability of the kit by using the stabilizer consisting of the four components on the basis of ensuring the detection accuracy, linear correlation and analysis sensitivity, and is beneficial to further popularization of the kit in the market.

Drawings

FIG. 1 is a graph showing the correlation between the kit of example 1 and the kit of comparative example 1;

FIG. 2 is a graph of the correlation of the kit of example 2 with the kit of comparative example 1;

FIG. 3 is a graph of the correlation of the kit of example 3 with the kit of comparative example 1;

FIG. 4 is a method for detecting the reagent kit of the present invention on a fully automatic biochemical analyzer.

Detailed Description

In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, the following detailed description is made with reference to specific embodiments and accompanying drawings.

Example 1:

the kit for detecting fibrin and fibrinogen degradation products comprises a reagent R1, a reagent R2 and a calibrator, wherein:

the components and contents of the reagent R1 are as follows:

Tris-HCL buffer 50mmol/L

Proclin300 3ml/L；

The components and contents of the reagent R2 are as follows:

the detection method comprises the following steps: the following operations were performed using a fully automatic biochemical analyzer (e.g., Hitachi 7180 fully automatic analyzer, OLYMPUS AU640, etc.):

the calibration material used by the invention is the calibration material produced by Beijing Jiuqiang biology company.

3ul of saline, sample or calibrator and 100ul of reagent R1 were added and incubated for 5min, the absorbance value A1 was recorded, then 100ul of reagent R2 was added and incubated for 5min, the absorbance value A2 was recorded, and finally Δ A was calculated.

Example 2:

the components and contents of the reagent R1 are as follows:

Tris-HCl buffer 30mmol/L

Proclin300 1ml/L；

The components and contents of the reagent R2 are as follows:

the detection method was the same as that of example 1, and the calibrator was prepared by Beijing Jiuqiang Biometrics.

Example 3:

the components and contents of the reagent R1 are as follows:

buffer solution 100mmol/L

Triton X-100 5ml/L；

The components and contents of the reagent R2 are as follows:

Comparative example 1:

the detection kit for fibrin and fibrinogen decomposition products of a certain company which is approved by the national food and drug administration commonly on the market is adopted.

Comparative example 2:

the components and contents of the reagent R1 are as follows:

Tris-HCL buffer 50mmol/L

Proclin300 3ml/L；

The components and contents of the reagent R2 are as follows:

Tris-HCL buffer 50mmol/L

0.1 percent of mouse anti-human FDP monoclonal antibody latex particles

Proclin300 3ml/L。

Comparative example 3:

the components and contents of the reagent R1 are as follows:

Tris-HCL buffer 50mmol/L

Proclin300 3ml/L；

The components and contents of the reagent R2 are as follows:

comparative example 4:

the components and contents of the reagent R1 are as follows:

Tris-HCL buffer 50mmol/L

Proclin300 3ml/L；

The components and contents of the reagent R2 are as follows:

and (3) accuracy test:

the comparative test was performed using the kits of examples 1 to 3 as experimental groups and the kit of comparative example 1 as a control group, and 40 samples were tested, and the test results are shown in FIGS. 1 to 3.

As can be seen from the detection data in fig. 1 to 3, the linear correlation coefficients r of the detection kits of examples 1 to 3 and the detection kit of comparative example 1 are respectively 0.9989, 0.9988, and 0.9989, and the correlation is greater than 0.99, which indicates that the kit of the present invention has high consistency with the detection kits for fibrin and fibrinogen decomposition products, which are approved in the market, and proves that the accuracy of the kit of the present invention is not affected by the addition of other components, and the kit of the present invention still maintains good accuracy.

Assay sensitivity test:

samples with known FDP concentrations of 8. mu.g/mL were tested using the kits of examples 1-3 and comparative examples 1-4 and the rate of change of absorbance (. DELTA.A/min) was recorded. The results are shown in Table 1.

TABLE 1 assay sensitivity test results for each kit

Theoretical concentration (μ g/mL)	8
		Example 1 measurement result Δ A/min	0.0565
Example 2 measurement results Δ A/min	0.0577
		Example 3 measurement result Δ A/min	0.0573
Comparative example 1 detection result DeltaA/min	0.0536

According to detection data, the components of the stabilizer added in the kit do not influence the analysis sensitivity of the kit, and the kit still maintains better analysis sensitivity.

And (3) stability test:

the stability of the reagent kit of example 1-3 and the reagent kit of comparative example 1-4 were tested by storing the reagents in a dark environment without corrosive gases at 2-8 ℃. The same sample is selected for each kit to be measured three times per month, an average value is taken, and the average value is compared with the detection result of the fresh reagent of the comparative example 1, so that the stability time of the reagent is determined. The test data are shown in Table 2.

TABLE 2 stability test results

The experimental result shows that the detection kit of the embodiment 1-3 is stable when stored in a light-shielding environment without corrosive gas at the temperature of 2-8 ℃ for 15 months, the reagent of the comparative example 1-4 is stable when stored in a light-shielding environment without corrosive gas at the temperature of 2-8 ℃ for 12 months, and the stability is reduced by nearly 80% at the time of 15 months. Therefore, the stability of the reagent is effectively improved by adding the stabilizer consisting of the four components of BSA, sorbitan fatty acid, cocamidopropyl betaine and polyvinylpyrrolidone into the reagent R2.

The foregoing is a preferred embodiment of the present invention, and it should be noted that modifications and adaptations can be made by those skilled in the art without departing from the principle of the present invention, and should be considered as within the scope of the present invention.

Claims

1. A fibrin and fibrinogen degradation product detection kit is characterized by comprising a reagent R1, a reagent R2 and a calibrator, wherein:

the components and contents of the reagent R1 are as follows:

buffer solution 30-100 mmol/L

11-5 ml/L of surfactant;

the components and contents of the reagent R2 are as follows:

2. the fibrin and fibrinogen degradation product detection kit of claim 1, wherein the buffer is Tris-HCL buffer and the PH is 8.0.

3. The fibrin and fibrinogen degradation product detection kit of claim 1, wherein the betaine is cocamidopropyl betaine.

4. The fibrin and fibrinogen degradation product detection kit of claim 1, wherein the surfactant 1 and the surfactant 2 are both Triton X-100, polyoxyethylene lauryl ether or a combination thereof.

5. The fibrin and fibrinogen degradation product detection kit of claim 1, wherein the anti-human FDP monoclonal antibody latex particles comprise one or more of mouse anti-human FDP monoclonal antibody, rabbit anti-human FDP monoclonal antibody, and goat anti-human FDP monoclonal antibody.

6. The fibrin and fibrinogen degradation product detection kit according to any one of claims 1 to 5, wherein the reagent R1 has the following components and contents:

buffer 50mmol/L

13 ml/L of surfactant;

the components and contents of the reagent R2 are as follows:

7. the fibrin and fibrinogen degradation product detection kit of claim 6, wherein the volume ratio of the reagent R1 to the reagent R2 is 1: 1.