Vacuum hemostix

A technology of vacuum blood collection tubes and tubes, applied in fungi, sensors, diagnosis, etc., to achieve the effect of reducing blood collection, improving work efficiency, and heavy workload

Inactive Publication Date: 2010-12-08
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] Synthesize the DNA sequence shown in Sequence 2 in the Sequence Listing using a fully artificial synthesis method, with recognition sites for restriction endonucleases EcoRI and BamHI at both ends of the sequence, and the DNA shown in Sequence 2 starts from position 262-459 at the 5' end bit encodes the protein of sequence 1 in the sequence listing. The synthesized DNA sequence was digested with EcoRI and BamHI, the digested fragments were recovered by electrophoresis, and stored at -20°C for future use.

[0021] b) Construction of vector pMPT-02

[0022] Using the genomic DNA of Hansenula polymorpha CGMCC 2.2497 as a template, clone the 1.5kb methanol oxidase gene (MOX) promoter, the 350bp methanol oxidase gene (MOX) terminator, and the 1.0kb self-replicating sequence HARS; and from YIp5 (GenBank Accession No.L09157) plasmi...

Embodiment 2

[0108] Example 2. Stability test of anticoagulant activity of general-purpose 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-purpose vacuum blood collection tubes is basically the same as the method for measuring the specific anticoagulant activity of hirudin. Only the abscissa of the regression line equation is changed from the mass of hirudin to the reciprocal of the volume of the diluent. See Table 1 for the results of the three determinations of the stability test of the anticoagulant activity of the general-purpose vacuum blood collection tube.

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

[0111] storage time

[0112] The blood collection volu...

Embodiment 3

[0113] Embodiment 3, the anticoagulant time of the universal vacuum blood collection tube of the present invention

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

[0115] Table 2, the relationship between the activity unit of recombinant hirudin and the anticoagulant time of the...

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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 gland of the European medical leech, and now it can be produced in large quantities by recombinant DNA technology. Hirudin is a single-chain polypeptide composed of 65 and 66 amino acid residues, with a molecular weight of 7,000Da. The mechanism of its inhibitory effect has been extensively studied. The hirudin molecule includes two functional regions: the N-terminal core region (1-52 residues) containing three disulfide bond structures and the random C-terminal tail (53-65 residues). Hirudin and thrombin form a very tight non-covalent reversible complex in a 1:1 ratio. The N-terminal core region binds to the hydrolysis active site of thrombin, while the C-terminal tail binds to the thrombin’s effect on fibrin. original recognition site. Both domains bind thrombi...

Claims

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Application Information

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