JP2000187032A - Method and device for separating blood serum - Google Patents

Abstract

PROBLEM TO BE SOLVED: To coagulate blood in a short time and to enable separation of blood serum containing no blood corpuscles without using a centrifugal separator by shaking a blood sampling tube to dissolve a specific blood corpuscle coagulant and a blood coagulation accelerator in blood, coagulating the blood, and moving a porous plate from the liquid level of the coagulated blood in the direction of the bottom of the tube. SOLUTION: Blood is sampled into a blood sampling tube 2 in which a blood corpuscle coagulant and a blood coagulation accelerator are placed, and the blood sampling tube 2 is shaken to dissolve the blood corpuscle coagulant and the blood coagulation accelerator in the blood. Then it is left in a stationary state to coagulate the blood. A porous plate 5 of smaller diameter than the inner diameter of the blood sampling tube 2 is moved from the liquid level of the blood in the direction of the bottom of the tube 2 to remove blood corpuscles floating in serum 8 to separate the serum 8. A blood corpuscles coagulant to be used has no special limitation, and it is possible to name compounds with amino groups of molecular weight 450 or more, i.e., synthetic high-molecular compounds such as polyacrylamide, low-molecular compounds such as a polyamine acid with a constituent component of a basic amino acid such as poly-L-lysine, vancomycin.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method and an apparatus for separating serum. More specifically, the present invention relates to a serum separation method and apparatus capable of coagulating blood in a short time and separating serum containing no blood cells without using a centrifuge.

[0002]

BACKGROUND OF THE INVENTION Blood is an important subject of clinical biochemical tests. Biochemical tests on blood include those performed using whole blood, such as specific gravity of whole blood, red blood cell count, white blood cell count, platelet count, hemoglobin, and hematocrit, lipids such as total cholesterol and HDL, GOT, and GPT. , Γ-GTP,
Enzyme systems such as LDH, vital dyes such as bilirubin, electrolyte systems such as sodium, potassium and serum iron, urea nitrogen,
There are tests performed using serum such as nitrogen-containing components such as creatinine. In order to obtain serum as a material for clinical biochemical test, it is necessary to remove clotting factors such as red blood cells, white blood cells, platelets, fibrinogen which is a clotting protein, factor V and factor VIII from blood. Conventionally, centrifugation has been used to separate serum from blood. The centrifugal separation method uses a centrifuge to collect blood from a blood collection tube and uses centrifugal force to separate serum and blood cells.However, strong centrifugal force and 10 minutes or more are required for separating serum and blood cells. Because of the long time required, rapid measurement as an emergency test cannot be supported. In addition, blood collection tubes containing blood must be manually mounted on the centrifuge, and the number of blood collection tubes that can be mounted on the centrifuge is limited. It hinders automation of inspection and mass processing. Therefore, there is a need for a serum separation method and apparatus that can coagulate blood in a short time and separate serum containing no blood cells without using a centrifuge.

[0003]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method and an apparatus for separating serum capable of coagulating blood in a short time and separating serum containing no blood cells without using a centrifuge. It was made for the purpose of.

[0004]

Means for Solving the Problems The present inventors have conducted intensive studies in order to solve the above-mentioned problems, and as a result, collected blood into a blood collection tube containing a hemagglutinating agent and a blood coagulation promoter, and After coagulating the blood, a perforated plate having a diameter smaller than the inner diameter of the blood collection tube is moved from the liquid surface of the coagulated blood toward the tube bottom, thereby pushing down the blood cells together with the fibrin net toward the tube bottom, and the serum is removed from the supernatant. And found that the present invention was completed based on this finding. That is, according to the present invention, (1) blood is collected in a blood collection tube containing a hemagglutinating agent and a blood coagulation promoting agent, and the blood collection tube is shaken to dissolve the hemagglutinating agent and the blood coagulation promoting agent in the blood; Blood coagulation by removing the blood cells from the serum by moving a perforated plate having a diameter smaller than the inner diameter of the blood collection tube from the liquid surface of the coagulated blood toward the tube bottom. Serum separation method,
(2) The serum separation method according to (1), wherein the hemagglutinating agent is a compound having an amino group and a molecular weight of 450 or more, (3)
The serum separation method according to (2), wherein the compound having an amino group and a molecular weight of 450 or more is a polyamino acid containing a basic amino acid as a constituent, and (4) the polyamino acid containing a basic amino acid as a constituent is a polyamino acid. (3) which is -L-lysine
(5) the serum separation method according to (2), wherein the amino group-containing compound having a molecular weight of 450 or more is vancomycin or kanamycin; (6) the blood coagulation promoting agent is a blood coagulation system; No. that is an endoprotease
(1) The serum separation method according to (1), (7) the serum separation method according to (6), wherein the endoprotease of the blood coagulation system is thrombin, and (8) the serum separation method according to (1), wherein the blood coagulation promoter is dextran. (9) The method for separating serum according to item (1), wherein the perforated plate has a network structure.
(10) A mechanism for opening a blood collection tube containing blood coagulated in the presence of a hemagglutinating agent and a blood coagulation promoter, and moving a perforated plate having a diameter smaller than the inner diameter of the blood collection tube from the liquid surface toward the tube bottom. A serum separating device comprising: a moving mechanism; and a separating mechanism for sucking serum at the upper part of the perforated plate.
1) The serum separating apparatus according to (10), wherein the perforated plate is moved from the liquid level to the bottom of the tube by mechanically pushing down the perforated plate. (12) The perforated plate is made of a ferromagnetic material, The serum separating apparatus according to (10), wherein the plate is moved from the liquid level toward the bottom of the tube by applying a magnetic field to the perforated plate from outside the blood collection tube, and (13) the perforated plate is formed of a mesh. It is intended to provide the serum separating device according to the above (10), which has a structure.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION In the serum separation method of the present invention, blood is collected in a blood collection tube containing a hemagglutinating agent and a blood coagulation promoter, and the blood collection tube is shaken to give the blood a hemagglutinating agent and a blood coagulation promoter. After dissolving the agent, let it stand and allow the blood to coagulate,
By moving the perforated plate having a diameter smaller than the inner diameter of the blood collection tube from the liquid surface of the coagulated blood toward the tube bottom, blood cells floating in the serum are removed from the serum to separate the serum.
There is no particular limitation on the hemagglutinating agent used in the method of the present invention, for example, polyalkylene oxide, polyalkyleneimine,
Examples thereof include compounds having an amino group and a molecular weight of 450 or more. Of these, compounds having an amino group and a molecular weight of 450 or more can be suitably used because they are easy to handle. Molecular weight of 45 having amino group
Examples of the 0 or more compounds include synthetic polymer compounds such as polyacrylamide, poly-L-lysine, and poly-L.
-Polyamino acids having a basic amino acid such as arginine as a component, and low molecular weight compounds such as vancomycin and kanamycin. Among these, polyamino acids having a basic amino acid as a component, particularly poly-L-lysine, vancomycin and kanamycin are:
Since it has a large cohesive force on blood cells, it can be suitably used. When poly-L-lysine is used as a hemagglutinating agent, its molecular weight is preferably relatively large,
More preferably, it is 30,000 to 200,000. The blood coagulation promoter used in the method of the present invention is not particularly limited, and examples thereof include a blood coagulation endoprotease and dextran. Examples of the blood coagulation endoprotease include thrombin, prothrombin, coagulation factor X, coagulation factor V and an activator thereof. Among them, thrombin can be used particularly preferably because it is easy to handle and has a large blood coagulation promoting power.

[0006] In the method of the present invention, the amounts of the hemagglutinating agent and the blood coagulation promoting agent to be previously placed in the blood collection tube are not particularly limited, and can be appropriately selected according to the respective hemagglutinating power and blood coagulation promoting power. . For example, when the hemagglutinating agent is poly-L-lysine, the amount is preferably 0.1 to 10 mg, preferably 0.5 to 5 mg per ml of blood.
Is more preferable. If the amount of poly-L-lysine is less than 0.1 mg per 1 ml of blood, the separated blood serum may be turbid due to insufficient hemagglutinating power. Poly-L
-The amount of lysine is usually 10 mg or less per 1 ml of blood, and if it exceeds 10 mg per 1 ml of blood, it may cause a problem in clinical biochemical tests. When the blood coagulation promoter is thrombin, the amount is 1 ml of blood.
Is preferably 0.01 to 5 units, and 0.1
It is more preferable that the number is 1 to 1 unit. If the amount of thrombin is less than 0.01 unit per 1 ml of blood, there is a possibility that the ability to promote blood coagulation is insufficient and the formation of separated serum is insufficient. When the amount of thrombin exceeds 5 units per ml of blood, clot formation is accelerated, but adhesion to the blood collection tube wall becomes strong, and there is a possibility that the thrombin may not be easily retracted by the perforated plate. In the method of the present invention, there is no particular limitation on the method of collecting blood into a blood collection tube containing a hemagglutinating agent and a blood coagulation promoter, and blood can be collected by a normal vacuum blood collection method. Blood is collected in a blood collection tube containing a hemagglutinating agent and a blood coagulation promoting agent, and the blood collection tube is shaken. And begin to settle toward the bottom of the tube. When only a blood coagulant is added to a blood collection tube and a blood coagulation promoter is not added, blood cells aggregate, but red blood cells adhere to the wall of the blood collection tube, and serum separation cannot be achieved in accordance with the purpose. By adding a blood cell aggregating agent and a blood coagulation promoter to the blood collection tube, the blood cell aggregate in the supernatant layer becomes large, and the amount of red blood cells adhering to the tube wall can be reduced.

In the method of the present invention, a blood cell agglutinating agent and a blood coagulation promoting agent are added to a blood collection tube, so that agglutination of blood cells becomes large and blood coagulates and sediments toward the tube bottom. A small amount of blood cells still float near the surface, and a small amount of blood cells still adhere to the tube wall. In the method of the present invention, in this state, the perforated plate having a diameter smaller than the inner diameter of the blood collection tube is moved from the surface of the coagulated blood toward the tube bottom. FIG. 1 (a) is a perspective view of one embodiment of a perforated plate used in the method of the present invention, and FIG. 1 (b) is a perspective view of another embodiment of a perforated plate used in the method of the present invention. The perforated plate of the embodiment shown in FIG. 1 (a) is a combination of a mesh-like mesh structure and an annular frame. The perforated plate is dropped on the surface of coagulated blood in a blood collection tube, and a rod or the like is placed from above. By mechanically depressing the perforated plate, or when the perforated plate is made of a ferromagnetic material, by applying a magnetic field from outside the blood collection tube using a magnet, etc., from the blood surface to the bottom of the tube. Can be moved. The perforated plate of the embodiment shown in FIG. 1 (b) is obtained by attaching a handle to a perforated plate having a network structure composed of mesh-like holes, and by mechanically pushing down the perforated plate using the handle. Can be moved from the liquid level of the coagulated blood toward the tube bottom. In the method of the present invention, the material of the perforated plate is not particularly limited. For example, a perforated plate made of a metal such as stainless steel can be used, or a perforated plate made of a plastic such as polypropylene can be used. By using a metal perforated plate, the regression effect of blood clots can be promoted. Perforated plates made of plastic can be easily mass-produced.
In the method of the present invention, the shape of the holes in the perforated plate is not particularly limited, and a perforated plate having holes such as a triangular, hexagonal, circular, or elliptical shape in addition to the network structure can be used. By moving the perforated plate from the liquid surface of the coagulated blood toward the tube bottom, the minute blood cells floating near the liquid surface and the minute blood cells adhering to the tube wall are not only red blood cells, but also white blood cells and platelets, fibrin. Entangled with the net, it is pushed down toward the bottom of the tube by the perforated plate and removed from the serum. As a result, a transparent serum containing no blood cells is obtained on the upper part of the perforated plate, and can be directly used for clinical biochemical examination. In the method of the present invention,
In addition to thrombin, calcium ions can be added as needed. By adding calcium ions,
Thrombin can enhance the blood coagulation promoting action.

The serum separating apparatus of the present invention comprises a mechanism for opening a blood collection tube containing blood that has been coagulated in the presence of a hemagglutinating agent and a blood coagulation promoter, and a liquid perforated plate having a diameter smaller than the inner diameter of the blood collection tube. It has a moving mechanism for moving from the surface to the bottom of the tube, and a separating mechanism for sucking the serum on the upper part of the perforated plate. FIG. 2 is an explanatory diagram of one embodiment of the serum separation device of the present invention. As shown in FIG. 2A, a blood collection tube 2 containing blood 1 coagulated in the presence of a hemagglutinating agent and a blood coagulation promoter.
Is removed by the lid opening mechanism 4. Then, FIG.
As shown in (b), a steel perforated plate 5 having a diameter smaller than the inner diameter of the blood collection tube is dropped on the liquid surface, and is moved from the liquid surface toward the tube bottom by a magnet 6 used as a moving mechanism. The red blood cells, white blood cells, and platelets in the coagulated blood are entangled with the fibrin network to form a blood clot 7, which is pushed down toward the tube bottom by the perforated plate and removed from the serum. As a result, a transparent serum 8 containing no blood cells is separated from the upper part of the perforated plate. The serum on the upper part of the perforated plate is aspirated by the separation mechanism 9 as shown in FIG. FIG. 3 is an explanatory diagram of another embodiment of the serum separation device of the present invention. As shown in FIG. 3A, the lid 3 of the blood collection tube 2 containing the blood 1 coagulated in the presence of the hemagglutinating agent and the blood coagulation promoter is removed by the lid opening mechanism 4. Then, FIG.
As shown in (b), a patterned perforated plate 10 having a diameter smaller than the inner diameter of the blood collection tube is used as a moving mechanism 11.
Mechanically depressed, and moves from the liquid level to the tube bottom. The red blood cells, white blood cells, and platelets in the coagulated blood are entangled with the fibrin network to form a blood clot 7, which is pushed down toward the tube bottom by the perforated plate and removed from the serum. as a result,
At the top of the perforated plate, a clear serum 8 containing no blood cells is separated. The serum on the upper part of the perforated plate is sucked by the separation mechanism 9 as shown in FIG.

[0009]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, which should not be construed as limiting the present invention. Example 1 Poly-L-lysine [Molecular weight 130,000-210,00
5 mL of human blood (male, 28 years old) in a blood collection tube [Eiken Tube No. 2] containing 5.0 mg of 0, Lot87H5913, Sigma] and 2.5 units of thrombin [Bovine Plasma, Green Cross Co., Ltd.] Was collected by vacuum blood collection. The blood collection tube was shaken to dissolve poly-L-lysine and thrombin in the blood, and the blood collection tube was allowed to stand. The blood clotted within 5 minutes. With the lid of the blood collection tube removed, 8 of the shape shown in FIG.
A steel perforated plate having a diameter of 8 mm in which a mesh net and an annular frame were combined was dropped on the liquid surface, and a magnetic force was applied to the perforated plate from the outside of the blood collection tube using a horseshoe-shaped magnet to move the perforated plate toward the bottom of the tube. The clot that had clotted and retracted was pushed down together with the perforated plate, and a clear serum without turbidity was separated on the top of the perforated plate. This serum was collected by suction, and could be directly used for clinical biochemical examination. Example 2 Except that the mesh of the steel perforated plate was 16 mesh,
The same operation as in Example 1 was repeated. When the perforated plate was moved toward the bottom of the tube, the clot that had clotted and retracted was pushed down together with the perforated plate, and clear serum without turbidity was separated at the top of the perforated plate. This serum was collected by suction, and could be directly used for clinical biochemical examination. Example 3 The same operation as in Example 1 was repeated, except that a perforated plate with a handle made of polypropylene having a diameter of 8 mm and having the shape shown in FIG. 1B was used and pressed down mechanically. When the perforated plate was pushed down, the clot that had clotted and retracted was pushed down together with the perforated plate, and a clear serum without turbidity was separated at the top of the perforated plate. This serum was collected by suction, and could be directly used for clinical biochemical examination. Example 4 The same operation as in Example 1 was repeated, except that the amount of thrombin put into the blood collection tube was 0.05 unit. When the perforated plate was moved toward the bottom of the tube, the clot that had clotted and retracted was pushed down together with the perforated plate, and a slightly turbid serum was separated at the top of the perforated plate. The serum was centrifuged for 1,000 hours.
After centrifugation at 0 rpm for 1 minute, the turbidity disappeared and could be subjected to clinical biochemical examination. Example 5 The same operation as in Example 1 was repeated except that the amount of thrombin to be put into the blood collection tube was 25 units. When the perforated plate was moved toward the bottom of the tube, the clot that had clotted and retracted was pushed down together with the perforated plate, and clear serum without turbidity was separated at the top of the perforated plate. This serum was collected by suction, and could be directly used for clinical biochemical examination.

[0010]

According to the method and apparatus for separating serum of the present invention, blood can be coagulated in a short time and serum free from blood cells can be separated without using a centrifugal separator. It is possible to easily automate all the steps of the clinical biochemical test, and to separate the serum in a short time, so that it is possible to cope with a rapid measurement as an urgent test.

[Brief description of the drawings]

FIG. 1 is a perspective view of two embodiments of a perforated plate used in the method of the present invention.

FIG. 2 is an explanatory diagram of one embodiment of the serum separation device of the present invention.

FIG. 3 is an explanatory view of another embodiment of the serum separation device of the present invention.

[Explanation of symbols]

 Reference Signs List 1 coagulated blood 2 blood collection tube 3 lid 4 lid opening mechanism 5 perforated plate 6 magnet 7 blood clot 8 serum 9 separation mechanism 10 perforated plate 11 handle

Claims (13)

[Claims] 1. Blood is collected in a blood collection tube containing a hemagglutinating agent and a blood coagulation promoting agent, and the blood collection tube is shaken to dissolve the hemagglutinating agent and the blood coagulation promoting agent in the blood. Serum separation, wherein blood cells floating in serum are removed from the serum by moving a perforated plate having a diameter smaller than the inner diameter of the blood collection tube from the liquid surface of the coagulated blood toward the tube bottom. Method. 2. The hemagglutinating agent has a molecular weight of 4 having an amino group.
2. The method for separating serum according to claim 1, which is 50 or more compounds. 3. The serum separation method according to claim 2, wherein the compound having an amino group and a molecular weight of 450 or more is a polyamino acid containing a basic amino acid as a constituent. 4. The method according to claim 3, wherein the polyamino acid comprising a basic amino acid is poly-L-lysine. 5. The compound having an amino group and a molecular weight of 450 or more is vancomycin or kanamycin.
The method for separating serum according to the above. 6. The method according to claim 1, wherein the blood coagulation promoter is a blood coagulation endoprotease. 7. The method according to claim 6, wherein the blood coagulation endoprotease is thrombin. 8. The method according to claim 1, wherein the blood coagulation promoter is dextran. 9. The method according to claim 1, wherein the perforated plate has a network structure. 10. A mechanism for opening a blood collection tube containing blood coagulated in the presence of a hemagglutinating agent and a blood coagulation promoter, and a perforated plate having a diameter smaller than the inner diameter of the blood collection tube from the liquid surface to the tube bottom. A serum separating apparatus comprising: a moving mechanism for moving; and a separating mechanism for sucking serum on an upper portion of a perforated plate. 11. Movement of the perforated plate from the liquid level toward the bottom of the tube,
11. The method according to claim 10, wherein the pressing is performed by mechanically pressing down the perforated plate.
The serum separation device according to the above. 12. The serum according to claim 10, wherein the perforated plate is made of a ferromagnetic material, and the perforated plate is moved from the liquid surface toward the tube bottom by applying a magnetic field to the perforated plate from outside the blood collection tube. Separation device. 13. The serum separating apparatus according to claim 10, wherein the perforated plate has a network structure.