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Vacuum hemostix

A vacuum blood collection tube, vacuum tube technology, applied in fungi, sensors, diagnosis and other directions, to achieve the effect of extending shelf life, large workload and improving work efficiency

Inactive Publication Date: 2008-08-27
汪和睦
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to find an ideal universal anticoagulant, people have spent decades, unfortunately until now, in the world market, there is still no general-purpose vacuum blood collection tube for laboratory testing.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, the preparation of vacuum blood collection tube

[0018] 1. Preparation of Hirudin

[0019] a) Synthesis of Hirudin Gene

[0020] The DNA sequence shown in Sequence 2 in the sequence listing was synthesized by a fully artificial synthesis method. The two ends of the sequence have recognition sites for restriction endonucleases EcoRI and BamHI. The DNA shown in Sequence 2 is from the 5' end of the 262nd-459th position. bit encodes the protein of Sequence 1 in the Sequence Listing. The synthesized DNA sequences were digested with EcoRI and BamHI, and the digested fragments were recovered by electrophoresis, and stored at -20°C for later use.

[0021] b) Construction of vector pMPT-02

[0022] Using Hansenula polymorpha CGMCC 2.2497 genomic DNA as a template, clone 1.5kb methanol oxidase gene (MOX) promoter, 350bp methanol oxidase gene (MOX) terminator, 1.0kb autonomously replicating sequence HARS; and cloned from YIp5 The 1.1kb Saccharomyces cerevisiae u...

Embodiment 2

[0108] Example 2. Stability test of anticoagulant activity of general vacuum blood collection tubes.

[0109] The prepared universal vacuum blood collection tubes were stored at 4°C and room temperature, respectively, and their anticoagulant activity was measured at different times. The method for measuring the anticoagulant activity of general vacuum blood collection tubes is basically the same as the method for measuring the anticoagulant specific activity of hirudin, except that the abscissa of the regression line equation is changed from the mass of hirudin to the inverse of the volume of the diluent. Table 1 shows the results of the three determinations of the stability test of the anticoagulant activity of the universal vacuum blood collection tube.

[0110] Table 1. Determination of Anticoagulant Activity Units of Recombinant Hirudin Universal Vacuum Blood Collection Tube

[0111] storage time

[0112] The above-mentioned universal vacuum blood co...

Embodiment 3

[0113] Example 3. Anticoagulation time of the universal vacuum blood collection tube of the present invention

[0114] The preset activity units of recombinant hirudin in blood collection tubes are 100, 200, 400, 800 and 1600 ATU / ml blood volume; three healthy subjects are named 1#, 2# and 3# respectively, and each person collects 5 tubes of blood , each tube of 2ml, the application of Japan Optoelectronics Celltac MEK-6318 automatic blood analyzer (white blood cells 5 classification), automatic detection of blood cells. There is an obvious dose-effect relationship between the different activity units of the preset recombinant hirudin and the anticoagulation time of the blood collection tube, that is, the higher the activity unit of the recombinant hirudin, the longer the anticoagulation time of the blood collection tube. The results are shown in Table 2:

[0115] Table 2. The relationship between the active unit of recombinant hirudin and the anticoagulation time of blood co...

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Abstract

The invention discloses a vacuum test tube which comprises a vacuum tube containing hirudin as anticoagulant therein. The hirudin can be in the form of freeze-dried powder or spray-dried powder or aqueous solution, or modified with polyethylene glycol or encapsulated in liposome. The content of the hirudin in the vacuum tube is 100-2000 ATU per milliliter of blood. The hirudin has an amino acid sequence (see sequence 1 in the sequence table). The vacuum test tube of the invention is used in clinical biochemistry, clinical immunology, clinical hemorheology, immune prototype analysis, RT-PCR detection and clinical hematological examinations (except cruor parameters) and realizes versatility of laboratory examinations.

Description

technical field [0001] The invention relates to a vacuum blood collection tube. Background technique [0002] Hirudin is a specific direct inhibitor of thrombin. Hirudin was originally purified from the salivary glands of the European medicine leech, and can now be produced in large quantities by recombinant DNA technology. Hirudin is a single-chain polypeptide consisting of 65 and 66 amino acid residues with a molecular weight of 7,000 Da. Much research has been done on the mechanism of its inhibitory effect. The hirudin molecule includes two functional regions: an N-terminal core region (1-52 residues) containing three disulfide bonds and a random C-terminal tail (53-65 residues). Hirudin forms a very tight non-covalent reversible complex with thrombin in a 1:1 manner, the N-terminal core region binds to the hydrolytically active site of thrombin, and the C-terminal tail binds to thrombin to fibrin. original recognition site. The two functional domains bind to thrombin...

Claims

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Application Information

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IPC IPC(8): A61B5/154A61J1/06A61J3/00A61K38/58C12N1/16
Inventor 汪和睦孙长海王昌华杨珺赵忠信
Owner 汪和睦
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