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Method for preparing liquid fibrinogen (FIB) detection solution

A technology for fibrinogen and detection reagents, applied in biological testing, material inspection products, etc., can solve the problems of high price, error reduction, poor sensitivity, etc., and achieve the effects of convenient use, strong stability, and strong compatibility

Inactive Publication Date: 2009-10-21
SHANGHAI LONG ISLAND BIOTEC CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The determination process is affected by many factors, among which the quality of the reagent is the most critical factor. Due to the problems of unstable product quality and poor sensitivity of the current domestic fibrinogen detection reagents, the detection results are unstable. The results measured between laboratories are difficult to compare. Although WHO standard reference materials are used, it is still difficult to reduce the error to an acceptable level. Therefore, most of the reagents used in clinical laboratories are imported products, which are expensive

Method used

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  • Method for preparing liquid fibrinogen (FIB) detection solution
  • Method for preparing liquid fibrinogen (FIB) detection solution
  • Method for preparing liquid fibrinogen (FIB) detection solution

Examples

Experimental program
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Effect test

Embodiment 1

[0013] Example 1: Preparation of Liquid Fibrinogen (FIB) Detection Reagent

[0014] Liquid fibrinogen FIB detection reagent is composed of bovine thrombin reagent, product stabilizer and FIB buffer, and its preparation method is as follows:

[0015] 1. Preparation of product stabilizer

[0016] Weigh 5g of polyethylene glycol, 2.5g of bovine serum albumin, 1g of hydroxybutyrate toluene, and 1g of sodium azide in turn, dissolve them in 20mM pH7.5 Tris-HCl buffer, add 30mL of glycerin, and mix well until the appearance is free. Color transparent liquid, dilute to 1L, store at 4°C for later use.

[0017] 2. Preparation of Bovine Thrombin Reagent

[0018] The bovine thrombin freeze-dried powder (500U / mL, 1mL / bottle) was diluted proportionally with a product stabilizer to a final concentration ≥ 100U / mL, and stored at 4°C for later use.

[0019] 3. Preparation of FIB buffer

[0020] Weigh 4.97g barbital sodium and 8.77g sodium chloride and add appropriate amount of distilled wa...

Embodiment 2

[0026] Embodiment two: the detection method of application liquid fibrinogen (FIB) detection reagent

[0027] 1. Drawing of Fibrinogen Standard Curve

[0028] Dilute the fibrinogen standard (3000mg / dL) with FIB buffer according to the ratio of 1:5, 1:10, 1:15, 1:20, 1:30 to the final concentration of 600mg / dL, 300mg / dL respectively , 200mg / dL, 150mg / dL, 100mg / dL, take 200uL of standard plasma with different concentrations, pre-warm at 37°C for 3 minutes, then add 100uL of thrombin reagent (no pre-warming), and use CA530 hemagglutination instrument ( Japan Sysmex company product) measure coagulation time. Take the fibrinogen standard substance content (mg / dL) of different concentrations as the abscissa, and take the corresponding coagulation time as the ordinate, draw the standard curve, and the correlation coefficient R of the standard curve 2 ≥0.99.

[0029] 2. Detection method

[0030] Dilute the plasma to be tested 1:10 with buffer, take 200uL of the plasma to be tested...

Embodiment 3

[0031] Embodiment three: Stability detection of liquid fibrinogen (FIB) detection reagent

[0032] The stability test of the reagent of the present invention includes stability at 2-8°C after opening, stability at 2-8°C without opening, accelerated destruction test at 37°C without opening, and long-term stability test.

[0033] After opening, store at 2-8°C, and within 30 days, the measured value of L-1 in normal quality control plasma is within the normal range with little change. The experimental data are shown in Table 1.

[0034] Table 1 Stability testing results of reagents of the present invention under storage conditions at 2-8°C

[0035]

[0036] Store the reagent at 2-8°C for 12 months, and test the stability of the reagent every other month. The testing instrument is a CA530 hemagglutination analyzer, and the commercially available APTT reagent matched with the instrument is used as a control. The plasma used It is the normal quality control plasma L-1 of Pacific...

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Abstract

The invention relates to a method for preparing liquid fibrinogen (FIB) detection solution. The solution is mainly used for monitoring the disturbance of blood coagulation, the hemorrhagic disease, the disseminated intravascular coagulation (DIC), the diabetes, the acidosis, the atherosclerosis and other disease symptoms. Along with the wide clinical application of thrombus and hemostasis detection, the fibrinogen (FIB) detection is the routine detection item of the thrombus and hemostasis detection, and has important clinical diagnosis value. The method adopts cow thrombin as a main raw material, and uses the Von clauss method (NCCLS recommended) to prepare the novel liquid fibrinogen (FIB) detection solution. The novel stable system solves the difficulty of maintaining the stability of the solution in the liquid state, and the method has the advantages of high sensitivity, convenient use, low experimental error, high stability, favorable compatibility, low cost, and the like.

Description

technical field [0001] The invention relates to a clinical diagnostic reagent, in particular to a preparation method of a liquid fibrinogen (FIB) detection reagent, and the method also includes the main components of the liquid fibrinogen (FIB) detection reagent. Background technique [0002] Fibrinogen (Fibrinogen, FIB) is a plasma globulin synthesized by the liver and is an important coagulation factor in the body. Its main physiological function is to participate in the coagulation process in vivo as coagulation factor I. Under the action of excess thrombin, fibrinogen in plasma will change from soluble protein to insoluble polymer, thus forming a fibrin clot. The thrombin clotting time of plasma is inversely proportional to the fibrinogen content. In the case of excess thrombin, the plasma clotting time is measured, and the fibrinogen content to be measured can be obtained by calculating the formula of the fibrinogen standard curve. [0003] In recent years, a large num...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 肖国伟
Owner SHANGHAI LONG ISLAND BIOTEC CO LTD
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