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Preparation method for liquid thrombin time (TT) detection reagent

A technology of thrombin time and detection reagents, which is applied in the direction of biological testing, material inspection products, etc., can solve the problems of high price, error reduction, poor sensitivity, etc., and achieve the effect of meeting the detection needs, convenient use, and strong compatibility

Inactive Publication Date: 2010-03-10
SHANGHAI LONG ISLAND BIOTEC CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The process of thrombin time detection and determination is affected by many factors, among which the quality of reagents is the most critical factor. Due to the problems of unstable product quality and poor sensitivity of current domestic coagulation reagents, the test results are unstable, and different clinical tests 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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  • Preparation method for liquid thrombin time (TT) detection reagent
  • Preparation method for liquid thrombin time (TT) detection reagent
  • Preparation method for liquid thrombin time (TT) detection reagent

Examples

Experimental program
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Effect test

Embodiment 1

[0012] Example 1: Preparation of liquid-type thrombin time (TT) detection reagent

[0013] The liquid type thrombin time (TT) detection reagent is composed of bovine thrombin reagent, product stabilizer and TT buffer, and its preparation method is as follows:

[0014] 1. Preparation of product stabilizer

[0015] Weigh 5g polyethylene glycol, 2.5g bovine serum albumin, 1g hydroxybutyrate toluene, 1g sodium azide in turn, dissolve them in 50mM pH7.5 Tris-HCl buffer, add 30mL glycerol, and mix well until the appearance is no Color transparent liquid, dilute to 1L, store at 4°C for later use.

[0016] 2. Preparation of Bovine Thrombin Reagent

[0017] The lyophilized bovine thrombin powder (500U / mL, 1mL / bottle) was diluted proportionally with the product stabilizer to a final concentration of 5U / mL, and stored at 4°C for later use.

[0018] 3. Preparation of TT Buffer

[0019] Weigh 6.06g of Tris base and 2.34g of sodium chloride and add an appropriate amount of distilled wat...

Embodiment 2

[0025] Embodiment 2: Detection method using liquid type thrombin time (TT) detection reagent

[0026] Take 100uL of plasma to be tested, pre-warm at 37°C for 3 minutes, then add 100uL of thrombin reagent (no pre-warming), and measure the clotting time with a CA530 coagulometer (product of Sysmex, Japan) or manually.

[0027] Precautions:

[0028] ①The sample avoids bubbles.

[0029] ②The specimen collection and coagulation test must use disposable plastic or siliconized glass containers.

[0030] ③ For the blood coagulation instrument measured by turbidity method, hemolysis and obvious jaundice and lipidemia will affect the measurement results. At this time, manual or electronic blood coagulation instrument should be used for measurement.

[0031] ④ When the hematocrit is outside the range of 20-55%, the dosage of anticoagulant should be adjusted. Anticoagulant dosage (mL)=0.00185*blood volume (mL)*(100-specific volume).

[0032] ⑤ Samples should not use oxalate, EDTA and ...

Embodiment 3

[0035] Example 3: Stability detection of liquid-type thrombin time (TT) detection reagent

[0036] The stability testing 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.

[0037] The L-1 value of normal quality control plasma was within the normal range within 30 days, and there was little change, and the experimental data were shown in Table 1.

[0038] The long-term stability test results of the reagent of the present invention under the storage condition of table 12-8°C

[0039]

[0040] The reagent was stored at 2-8°C for 12 months, and the stability of the reagent was tested every other month. It is the normal quality control plasma L-1 of Pacific Company, and the test results are shown in Table 2.

[0041] Table 2 Comparison of the stability of the liquid TT reagent of the present invention and the commercially ava...

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Abstract

The invention relates to a preparation method for a liquid thrombin time (TT) detection reagent, which also comprises main components of the liquid thrombin time (TT) detection reagent. The TT is a simple and convenient test for detecting functions of coagulation, anticoagulation and a fibrinolytic system, is widely used for monitoring and treating high molecular weight heparin and treating fibrinolysis, and for screening fibrinogen disorder and some fibrinogen deficiency symptoms and has significance on various clinic disease diagnoses. The preparation method for the liquid thrombin time (TT)detection reagent adopts a bovine thrombin as a main material and adopts a unique stabilization system to prepare a novel liquid thrombin time (TT) detection reagent. The novel stabilization system solves the difficult problem of maintaining the stability of a reagent in a liquid state, and has the advantages of high sensitivity, convenient use, small experiment error, strong stability, good 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 thrombin time (TT) detection reagent, which further comprises the main components of the liquid thrombin time (TT) detection reagent. Background technique [0002] Thrombin time (TT) is the time required for coagulation to be determined by adding a certain amount of thrombin reagent to the plasma to be tested, and fibrinogen becomes insoluble fibrin. TT is a simple test to detect the function of coagulation, anticoagulation and fibrinolysis system. It is widely used in monitoring and treatment of high molecular weight heparin and fibrinolysis treatment, screening of fibrinogen disorders and some serious fibrinogen deficiency diseases. The diagnosis of diseases in the department is of great significance. [0003] The principle of thrombin time (TT) detection is that an appropriate amount of thrombin converts fibrinogen in the plasma sample into fib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
Inventor 肖国伟
Owner SHANGHAI LONG ISLAND BIOTEC CO LTD
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