JPH0593721A - Serum/plasma separating apparatus - Google Patents

Abstract

PURPOSE:To obtain such an inexpensive serum/plasma separating apparatus that makes ordinary blood collecting operations and can nearly simultaneously, easily, and efficiently separate and recover a serum and plasma components which are usable for, for example, several to about ten inspection items from a collected whole blood sample during or after the sample blood is collected. CONSTITUTION:This serum/plasma separating apparatus is constituted of an outer tube 2 with a blood-collecting needle 1 protruded from the front end section of the tube 2 and inward needle 1a communicated with the needle 1 in the tube 2, blood-collecting tube 3 which is housed in the tube 2 in a slidable state and the front end side of which is closed with a plug body into which the needle 1a can be inserted, and a separated liquid collecting tube 5 which is communicable with the rear end side of the tube 3 through a hollow needle 4b and the inside of which is reduced in pressure. A blood corpuscle separating filter 6 is housed in the blood-collecting tube 3 at positions where the front and rear end sides are shielded.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a device for easily and efficiently separating and collecting serum / plasma components from a collected whole blood sample at the time of collecting blood from a human or an animal.

[0002]

2. Description of the Related Art Separation of serum or plasma components from whole blood has been common since a method using a centrifuge has been adopted. On the other hand, due to the progress of dry chemistry reagents, we can respond to emergency tests and rapid bedside tests.
Mechanisms have been developed that allow the separation of serum or plasma within the reagents themselves.

For example, Vogel et al., US Pat. No. 4,477,575, developed a method of using a test paper in which a blood cell separated portion of glass fiber and a reagent reaction portion are integrated. According to this, a trace amount (several μl) of serum or plasma can be separated from a total blood number of 10 μl, and a measured value can be obtained in real time. See also Miles (Mi
les) Japanese Patent Laid-Open No. 64-21362, a lectin having erythrocyte condensing activity and an absorptive material (hydrophilic inorganic powder cellulosic material etc.) are used to produce an absorptive matrix,
A method for separating blood cell components and serum / plasma components from whole blood using this is disclosed.

[0004]

According to the conventional separation method using a centrifuge, it is suitable for a large hospital or the like that tests a large number of specimens, but in a small-scale facility such as a small private hospital, Since such a centrifuge is expensive, it is hardly installed, and it is in a state where it is not enough to perform basic tests of several items to about 10 items. In such facilities, whole blood is stored as it is, and then clinical laboratory etc. Currently, we are asking for inspection. Therefore, the test result cannot be obtained in real time, and it sometimes takes several days. Further, since the whole blood is stored as it is, there is a case where an error occurs in the test value depending on the test item due to hemolysis or the like, and development of a simple separation method has been earnestly desired.

Also, Vogel et al., US Pat. No. 4,477,
The method disclosed in Japanese Patent No. 575 uses a test paper in which a blood cell separation part and a reagent reaction part are integrated. In this case, since the separated serum / plasma components are minute, naturally, It is impossible to use it for other inspection items and there is a drawback that it cannot be inspected other than the inspection items prepared by this measurement system. Further Miles JP-A-64-
The method of No. 21362 has not been put to practical use because blood cell separation is actually insufficient.

The present invention is intended to solve the above-mentioned drawbacks, and its purpose is to be carried out in the same manner as a normal blood collection operation, and almost simultaneously from whole blood samples collected during or after blood collection, for example, from several items. It is intended to provide an inexpensive serum / plasma separation instrument capable of easily and efficiently separating / collecting an amount of serum / plasma components that can be provided for about 10 items of inspection.

[0007]

The features of the present invention for solving the above-mentioned conventional problems and achieving the intended purpose are as follows.
An outer tube having a blood collection needle protruding outside the tip and having an inward needle communicating with the blood collection needle therein, and a plug body slidably accommodated in the outer tube and capable of being pierced by the inward needle at the tip side. A blood collection tube closed by means of a blood collection tube, and a separation liquid collection tube whose inside is decompressed and which is capable of communicating with the rear end side of the blood collection tube. It is in a serum / plasma separation device containing. The blood collection tube is in a state close to a vacuum with both ends closed (Claim 2)
Alternatively, a hollow needle may be provided at the rear end so as to communicate with the separated liquid collection tube (Claim 3), and the blood cell separation filter may be impregnated with a polyacrylic ester derivative and polyethylene glycol. It can be constituted by a blood cell separation fibrous material layer made of the above-mentioned glass fiber (Claim 4), and further, it can be constituted by integrating a lectin impregnated layer in which a fibrous base material is impregnated with cretin (Claim). Item 5).

[0008]

According to the serum / plasma separation device of the present invention, when a vacuum blood collection tube is used as in claim 2, the blood collection tube is pushed with the collection needle piercing the blood collection target, and the inward needle is inserted into the blood collection tube. Blood is collected in the blood collection tube by inserting the blood into the blood collection tube. After collecting the required amount of blood, pull out the blood collection needle and connect the separation liquid collection tube to the rear end of the blood collection tube, so that the blood in the blood collection tube is aspirated through the blood cell separation filter,
The serum and plasma components are separated by the filter and sucked into the separated liquid collection tube.

When a blood collection tube that is not in a vacuum state is used as in claim 3, the blood collection needle is stabbed into the blood collection tube while the blood collection tube is pushed and the inward needle is pierced into the blood collection tube. By connecting the separation liquid collection tube to the rear end of the blood collection tube, blood is drawn by the suction force from the separation liquid collection tube, which is sucked through the blood cell separation filter, and the separated serum / plasma is sucked into the separation liquid collection tube. To be done.

[0010]

Embodiments of the present invention will be described in detail below with reference to the drawings. 1 and 2 show a first embodiment of the present invention. The serum / plasma separation device shown in the figure is a bottomed cylindrical outer tube 2 having a blood collection needle 1 at the tip, a blood collection tube 3 fitted in the outer tube 2 and having a pressure reduced to a state close to a vacuum, and this blood collection tube. 3 is fitted to the rear end of the connector 3, and the separated liquid collection tube 5 is depressurized to a state close to a vacuum fitted to the rear end of the connector 4.
And have. In the non-collected state, the blood collection tube 2 and the connector 4
And the separated liquid collection tube 5 is in a separated state.

The outer tube 2 has a cylindrical shape with a bottom, and the blood sampling needle 1 is fixed by penetrating the tip of the outer tube 2 and the rear end thereof is an inward needle 1a.
Has become. The blood collection tube 3 has a cylindrical main body 3a whose both ends are open, and has a non-air-permeable plug 3b made of a rubber material at both ends.
It is configured to be closed with 3c, and the blood cell separation filter 6 is housed inside. The blood cell separation filter 6 has a two-layered structure including a blood cell separation fiber material layer 6a and a hemagglutination agent layer 6b impregnated with lectin. The hemagglutination agent layer 6b is not always necessary.

The connecting tool 4 is in the shape of a cylinder having a partition wall 4a at the substantially center thereof, and the connecting hollow needles 4b of both ends are fixed axially through the partition wall 4a. .. The separated liquid sampling tube 5 has a cylindrical shape with a bottom, and the opening thereof is closed by a non-air-permeable stopper 5a made of a rubber material to reduce the pressure inside to a substantially vacuum state.

Next, a method of using the serum / plasma separation device having the above configuration will be described. First, when collecting blood, the blood collection needle 1 is inserted into the subcutaneous blood vessel, and then the blood collection tube 3 is pushed in and the inward needle 1a is inserted into the plug body 3b. As a result, the blood is vacuum suctioned and flows into the blood collection tube 3. After collecting the required amount of blood in this way, the blood collecting needle 1 is pulled out from the skin. After that, when the separation liquid collecting tube 5 is pressed against the rear part of the blood collecting tube 3 via the connector 4, the connecting hollow needles 4b of both heads are respectively attached to the plug body 3c on the blood collecting tube 7 side and the separation liquid collecting tube 5 side. Break through the stopper 5a. As a result, the blood contained in the blood collection tube 3 passes through the blood cell separation filter 6 and is vacuum-sucked to the separated liquid collection tube 5 side. At this time, the blood passing through the blood cell separation filter 6 is separated into blood cells and serum / plasma components,
Only the serum / plasma components are collected in the separated liquid collecting tube 5.

It has been confirmed that several hundred μl of serum / plasma can be obtained from whole blood having a blood collection amount of about 1 ml when the collection operation is performed by this method, and at the same time as the blood collection, 10 or more blood samples are collected. It is possible to obtain a sufficient amount of serum / plasma capable of testing a large number of blood components across items.

The blood cell separation fiber material layer 6a of the blood cell separation filter 6 is preferably a glass fiber having an average diameter of 0.5 to 2.5 μm impregnated with a polyacrylic ester derivative and polyethylene glycol (PEG). As the polyacrylic ester derivative, polybutyl acrylate (PBA) and polymethyl acrylate (PMA) or polyethyl acrylate (PEA) are suitable.
As a mixing weight ratio with G, (10-12): (1-
4): (1-4), when the total content is 2-3%,
Good blood cell separation ability.

The hemagglutination agent layer 6b is formed by impregnating a non-woven fabric with a lectin which is a hemagglutination agent, and the hemagglutination agent may be any non-blood group specific lectin. The concentration is 0.01-0.005
A small amount of% had a sufficient effect. Lectins that can be used are, for example, Abrus precatoris (Aplin, Azuki beans), Bauhhinia purpurea (Mokwanju), Caragana.
arborescens (Siberian beans), Codium fragile (sea green algae), Concanavalin A (tachinata beans), Glycine max
(Soybean), Lathyrus odoratus (sweet pea), Lens
culinaris (lentils), Limulus polyphemus (horseshoe crab), Lycopersicon esculentum (tomato), Maclura
pomifera (Osage orange), Mycoplasma gallisept
icum, Perseau americana (Avocado), Phaseolus cocc
ineus, Phaseolus Vulg
aris (red kidney beans), Phytolaccaamericana (Pokeweed), Pisum sativum (peas), Psophoca
rpus tetragonolobus (wing bean), Ricinus co
mmunis (castor), Solanum tuberosum (potato), Tr
Iticum vulgaris (wheat malt), Vicia faba (broad beans), Vignaradiate (mung beans), Viscum album (European mistletoe), Wisteria floribunta (Fuji) and the like.

In the above-mentioned embodiment, the connecting tool 4 and the separated liquid collecting tube 5 are separated from the blood collecting tube 2, but these are combined in advance as shown in FIG. Alternatively, the blood collection tube 2 may be simultaneously pushed by pushing the separation liquid collecting tube 5 while the blood collecting needle 1 is being pierced into the blood vessel.

FIG. 4 shows a third embodiment of the present invention.
The rear end of the outer tube 2 is extended, and the separated liquid collecting tube 5 is slidably fitted to the rear portion of the outer tube in advance. In addition, a hollow needle 10 having one end previously penetrated through the rear end of the blood collection tube 3 protrudes, and the protruding end of the hollow needle 10 faces the separation and collection tube 5 side while being supported by a partition plate 11. is doing.

The end of the hollow needle 10 on the side of the sampling tube 3 is provided integrally with a filter support plate 10a that expands like a funnel, and the support plate 10a supports the rear part of the filter 6 so that it can move. To prevent. In this configuration, when the separated liquid sampling tube 5 is pushed during blood sampling, the inward needle 1
a, the hollow needle 10 breaks through the plugs 3b, 5a,
Blood flows into the blood collection tube 3 by the vacuum suction operation of the collection tube 5, and the serum separated through the filter 6
Plasma flows into the blood collection tube. Therefore, according to this embodiment, serum / plasma can be separated and collected simultaneously with blood collection. Further, the co-movement of the filter 6 during the vacuum suction is caused by the support plate 10.
This is prevented by a, and more efficient separation and sampling are performed.

[0020]

[Test example]

a) Preparation of blood cell separation layer material 1 g of fibrous glass was mixed with 24 mg of butyl polyacrylate and 4 methyl polyacrylate in 10 ml of methanol.
The solution prepared by dissolving mg and 2 mg of polyethylene glycol was sprayed, processed into an appropriate size shape, a sheet shape, a crushed small piece shape, a columnar shape, a cylindrical shape, etc., and dried at 100 ° C. to be manufactured. b) Preparation of serum or plasma separation filter The crushed piece-shaped material prepared above is used for an inner diameter of 7 mm and a height of 30 m.
10 mm column column, and glycine on top of it
Non-woven fabric "Kinocloth K70" (trademark) manufactured by Honshu Paper Manufacturing Co., Ltd., which is impregnated with 0.01% lectin physiological saline containing max (soybean) and dried at room temperature for about 1 hour, is cut into the same size (inner diameter 7 mm). Then, a filter was prepared by stacking.

A filter prepared without containing polybutyl acrylate, methyl polyacrylate and polyethylene glycol was separately adjusted to visually determine whether serum or plasma could be separated from 500 μl of whole blood. A remarkable separation ability was observed in the column prepared by containing butyl polyacrylate, methyl polyacrylate and polyethylene glycol. On the other hand, a filter containing no polybutyl acrylate, polymethyl acrylate and polyethylene glycol was judged to have no separability.

Next, the following tests were carried out using the above filters in order to see if the amount of components actually separated in serum or plasma was different from that in the case where the conventional centrifugation method was used.

Test Example 1 Serum Glucose Measurement Three types of specimens having different concentrations were used, one was subjected to serum separation by centrifugation (method 1), and the other was subjected to serum separation using the serum separation column of the present invention. (Methods 2), the glucose concentration of both was measured by a commercially available reagent (enzymatic method), and the results are shown in Table 1. As shown in Table 1, no difference between the separation methods was observed and good results were obtained.

Table 1 Number of times Sample I Sample II Sample III Method-1 Method-2 Method-1 Method-2 Method-1 Method-2 21 241 242 528 528 536 808 811 2 242 241 529 539 808 816 Unit: mg / dl test Example 2 Measurement of Serum 3-Hydroxybutyric Acid Using three types of specimens having different concentrations, one was subjected to serum separation by centrifugation (method-1), and the other was subjected to serum separation using the serum separation column of the present invention ( Method 2) and the commercially available reagent (enzymatic method) were used to measure the concentrations of both 3-hydroxybutyric acid, and the results are shown in Table 2. The results are as shown in Table 2.
There was no difference between the separation methods, and good results were obtained.

Table 2 Number of times Sample I Sample II Sample III Method-1 Method-2 Method-1 Method-2 Method-1 Method-2 21 89 91 109 109 110 187 186 2 90 93 112 112 110 185 185 Unit: μmol / l Test Example 3 Total Bile Acid Measurement Using three types of specimens having different concentrations, one was subjected to serum separation by centrifugation (method-1), and the other was subjected to serum separation using the serum separation column of the present invention (method- 2) The concentration of both bile acids was measured by a commercially available reagent (enzymatic method), and the results are shown in Table 3. As shown in Table 3, no difference in separation method was observed, and good results were obtained.

Table 3 Number of times Specimen I Specimen II Specimen III Method-1 Method-2 Method-1 Method-2 Method-1 Method-2 21 10.5 10.9 17.2 17.8 33.1 33.4 2 10.6 10.9 17.2 18.1 33.0 33.6 Unit: mg / dl

[0027]

As described above in detail with reference to the embodiments, according to the serum / plasma separation device of the present invention, blood cells and a testable amount of serum of 10 or more items are collected simultaneously with or immediately after blood collection. Since plasma can be separated and collected, the complexity of the conventional separation operation using a centrifuge is eliminated, and it can be separated and collected easily at low cost. Moreover, since the blood can be separated at the same time as or immediately after the blood collection, the measurement accuracy can be improved.

[Brief description of drawings]

FIG. 1 is a perspective view of a serum / plasma separation device according to a first embodiment of the present invention.

FIG. 2 is a sectional view of the same.

FIG. 3 is a sectional view of a serum / plasma separation device according to a second embodiment of the present invention.

FIG. 4 is a sectional view of a serum / plasma separation device according to a third embodiment of the present invention.

[Explanation of symbols]

 1 Blood Collection Needle 1a Inward Needle 2 Outer Tube 3 Blood Collection Tube 3a Main Body 3b, 3c, 5a Plug 4 Connector 4a Partition Wall 4b, 10 Hollow Needle 5 Separated Liquid Collection Tube 6 Blood Cell Separation Filter 6a Blood Cell Separation Fiber Material Layer 6b Blood Cell Flocculant layer 10a Filter support plate

Front page continuation (72) Inventor Hiroshi Okada 35, Higashi Sotobori-cho, Higashi-ku, Nagoya City, Sanwa Chemical Research Institute Co., Ltd. ) Inventor Masanori Kurono 35 Higashi Sotobori-cho, Higashi-ku, Nagoya City Sanwa Chemical Research Institute Co., Ltd.

Claims (5)

[Claims] 1. An outer tube having a blood collection needle protruding outside the tip and having an inward needle communicating with the blood collection needle therein, and a slidably housed inside the outer tube, and the inward needle pierced at the tip side. It is composed of a blood collection tube closed by a pluggable plug and a separated liquid collection tube whose inside is decompressed and which is capable of communicating with the rear end side of the blood collection tube, and shields the front and rear end sides inside the blood collection tube. A serum / plasma separation device that contains a blood cell separation filter in its arrangement. 2. The serum according to claim 1, wherein the blood collection tube is closed at both ends and is depressurized to a state close to a vacuum.
Plasma separation device. 3. The serum / plasma separation device according to claim 1, wherein the blood collection tube has a hollow needle projecting from the rear end thereof, and the hollow blood needle allows communication with the separation liquid collection tube. 4. The serum / plasma separation device according to claim 1, wherein the blood cell separation filter has a blood cell separation fiber material layer made of glass fibers impregnated with a polyacrylic ester derivative and polyethylene glycol. .. 5. The blood cell separation filter comprises a layer composed of glass fibers impregnated with a polyacrylic ester derivative and polyethylene glycol, and a hemagglutination agent in which a lectin-impregnated fiber base material is impregnated with lectin. The serum / plasma separation device according to claim 1, 2, or 3.