JPS59196084A - Device for treating blood - Google Patents

Abstract

PURPOSE:A device containing a solution having both a hemolysis agent and an anticoagulant, keeping a rubber stopper communicated with blood-containing space, being sterilized, capable of blending the blood with the solution, etc. sufficiently in a closed system. CONSTITUTION:The mixed solution 2 having a hemolysis agent, an anticoagulant, and, if necessary, a liquid medium, is contained in the container 1, the rubber stopper 3 is detachably attached to the opening of the container 1, and the container is hermetically sealed. The interior of the container 1 is kept at reduced pressure, so that collected blood is easily introduced into the container 1. The device for treating blood is sterilized in order to prevent the collected blood being infected with them.

Description

DETAILED DESCRIPTION OF THE INVENTION (1) Background of the Invention Technical Field The present invention relates to a novel blood processing device used for separating and culturing bacteria in blood.

In serious systemic infections such as sepsis and bacteremia, bacteria are present in the patient's blood, so when diagnosing these infections,
4 A blood bacteriological test is performed. Bacteria testing of blood is also carried out to determine the efficacy of antibacterial agents through animal testing. In these bacterial tests, it is first necessary to culture and isolate bacteria from sample blood, and then drug susceptibility tests and bacterial identification are performed. The instrument of the present invention is used for specimen processing during such bacterial testing.

(Prior Art and Problems) Conventionally, in the culture and isolation of blood bacteria, collected blood is mixed with a liquid nutrient medium and cultured until the medium becomes cloudy with bacteria grown in the medium, and then the grown bacteria are cultured on a blood agar plate. Colonies are grown by transplanting them onto a medium such as a chocolate agar plate and culturing them further.

In this way, with conventional methods, it is difficult to directly isolate and culture small numbers of bacteria from blood, and it is necessary to increase the number of bacteria through an additional operation called enrichment culture as a pretreatment for cultivating colonies. This requires complicated operations and a long time. In particular, enrichment culture in a liquid as described above generally requires a long time. In addition, blood itself has the effect of inhibiting the growth of bacteria, and when antibacterial agents are used as a membrane, it is even more difficult for bacteria to grow in the blood, and when the number of bacteria in the sample is small, Sometimes it is undetectable. Furthermore, when the bacteria are transferred from the growth medium to the culture medium, there is a risk that the test bacteria may contaminate the environment or contaminate the test bacteria, reducing the accuracy of the test.

■1 Purpose of the Invention Therefore, as a result of extensive research, the inventors of the present invention have found that bacterial ronin blood is mixed with a solution consisting of at least a hemolytic agent and an anticoagulant, or a solution consisting of a hemolytic agent, an anticoagulant, and a liquid medium. , a filter having pores large enough to prevent bacteria from passing through the mixture, and a water absorbent adhered to the back surface of the filter or a water absorbent impregnated with a liquid medium and dried, are housed in a container with the filter facing upward. We have developed a method for isolating and culturing bacteria in the blood by filtering it with a filtration incubator and culturing the bacteria on the filter to -1.

According to this culture method, it is possible to directly collect blood bacteria without the need for pretreatment operations such as enrichment culture of blood bacteria, and to culture them to that extent without transplanting them to another medium. . Therefore, in this culture method, it is essential to thoroughly mix the collected blood with a solution consisting of a hemolytic agent and an anticoagulant, or a solution consisting of a hemolytic agent, an anticoagulant, and a liquid medium. must be carried out in a closed container to prevent environmental contamination and specimen contamination by test bacteria.

Accordingly, an object of the invention is to provide an instrument that allows mixing a solution consisting of a hemolytic agent and an anticoagulant or a solution consisting of a hemolytic agent, an anticoagulant, and a liquid medium in a closed system, at least by aliquoting blood. .

The above objects are thus achieved by the device of the invention as described below.

(1) A blood processing device that is sterilized and contains a solution containing at least a hemolytic agent and an anticoagulant, and has a blood storage space. - (2) The blood processing device according to item 1, wherein the blood treatment solution contains food≠medium.

(3) The blood processing device according to item 1 or 2, wherein the blood storage space is under reduced pressure.

(4) The blood processing device according to item 1 or 2, wherein the container comprises a cylindrical body and a pusher arranged to slide on the inner wall of the cylindrical body in a liquid-tight manner.

By using the blood processing device of the present invention having the above structure, the blood can be treated as a hemolytic agent. It can be sufficiently mixed with a solution consisting of a hemolytic agent, an anticoagulant, or a solution consisting of a hemolytic agent, an anticoagulant, and a liquid medium in a closed system. There is no risk of contamination in the sample.

(2) Detailed Description of the Invention The configuration of the present invention will be described in detail below based on embodiments shown in the drawings.

FIG. 1 shows an embodiment of the blood processing device of the present invention.

This blood processing device consists of a container 1 containing at least a mixed solution 2 of a hemolytic agent and an anticoagulant, and a rubber stopper 3 removably and sealably fitted into the opening of the container 1. The interior of the is maintained at reduced pressure. To process blood using this blood processing device, the needle of a syringe (not shown) used to collect blood is pierced through the rubber stopper 3, and the blood is introduced into the container 1 using the reduced pressure inside the container 1. After the blood has been introduced, the needle is removed and the container is inverted to mix the blood with the solution in the container. Saponin is preferably used as the hemolytic agent, and sodium amylosulfate, sodium polyanethole fA acid, etc. are preferably used as the anticoagulant. A liquid medium can also be mixed with these mixed solutions, if desired. In this case, there is no need to impregnate the water absorbing body of the separation culture vessel with the medium to be used in the next operation. Also, bacteria multiply faster.

FIG. 2 shows another embodiment of the blood processing device of the present invention. This blood processing device consists of a cylindrical body 4 and a pusher 5 arranged to slide on the inner wall of the cylindrical body in a liquid-tight manner.A rubber plug 3 is removably attached to the opening of the cylindrical body 4. Discharged in a large and sealable manner,
A mixed liquid 2 consisting of a hemolytic agent and an anticoagulant similar to that of the previous embodiment is contained in the space formed by the cylindrical body 4 and the pusher 5.

This mixed solution may also be mixed with a liquid medium. To process blood using this blood processing device, insert the needle of a syringe that has collected 1d1 blood through the rubber stopper 3, and press the blood into the syringe. Blood is introduced into the container 83 while the pusher 5 of the processor slides downward. After the blood has been introduced, the container is inverted to thoroughly mix the blood with the hemolytic agent and anticoagulant. These blood processing devices are sterilized by γ-ray irradiation or autoclaving to prevent contamination of sample blood with bacteria.

By treating blood with the blood processor of the present invention, red blood cells are destroyed and blood coagulation is prevented.

These operations are essential for the subsequent blood filtration operation and culture of blood bacteria to be performed smoothly. In addition, if a liquid medium is mixed, a mixing process can be performed at the same time.

After treating the blood with the blood processor of the present invention, the rubber stopper is removed, and the sample blood is poured onto the filter of the separation incubator shown below.

As shown in FIG. 3, the filtration incubator consists of a container body 8 containing a filter 6 having holes large enough to prevent bacteria from passing through, and a water absorbent 7 bonded to the back side of the filter 6 with the filter 6 side up. It consists of a lid 9 that removably covers the opening of the container body 8.

The container body 8 includes a filter 6, a container body side wall 10, and a lid 9.
and a space 1 defined by the water absorbent body 7, the container body side wall 10, and the container body bottom 12.
3, and a ventilation hole 14 is provided below the water absorbent body 7.

The filter 6- is for filtering bacteria in the blood and has holes large enough to prevent bacteria from passing through. The pore size of the filter 6 is 0.75 microns or less, preferably 0.45 microns.
It is on the order of microns. The material of the filter 6 is not particularly limited as long as it is inert to blood, but representative examples include nitrocellulose, polycarbonate, polyamide, and cellulose ester.Commercially available materials include Millidea (product of Millipora Corporation). ) and Metricel (a product of Kelman Instrument Company). The filter 6 is desirably treated to be hydrophilic by a method known per se so that blood can easily absorb it.

The water absorbing body 7 absorbs and retains the filtered blood filtered through the filter 6, and provides nutrients to the bacteria captured on the Noilcro. It is desirable that the water absorbent body 7 has the ability to absorb almost the entire amount of filtrate.

Suitable materials include cellulose filter paper and nonwoven fabric. It is necessary that the water absorbent body 7 is closely fixed to the back surface of the filter 6 with an adhesive or the like, and if the adhesion is insufficient, filtration will not be performed smoothly, and the water absorbent body 7 will be tightly fixed to the back surface of the filter 6 after filtration. Nutritional supply to the bacteria on top is not sufficient.

As the adhesive used, heat sealing using low melting point polymer fibers that does not inhibit filtration is preferred. The water absorbent body 7 can also be impregnated with a liquid medium and dried if desired. In this case, the dry medium is dissolved in the liquid absorbed and retained by the water absorbent body 7, and provides nutrients and moisture to the bacteria on the filter 6.

The integrated filter 6 and water absorbent body 7 are fitted and fixed to the container body wall 10 without any gaps 9, thereby preventing sample blood from leaking out without passing through the filter 6.

The filter 6 and the water absorbent body 7 are fixed to the wall of the container body using an adhesive or by crimping with a caulking tool 15 as shown in the figure. In this case, a step S is formed on the side wall 10 of the container body 8, and a filter 6 integrated with this step S is formed.
After placing the water absorbent body 7, the caulking tool 15 may be press-fitted. A space 11 above the filter 6 is for storing sample blood, and a space 13 below the water absorbent body 7 is for receiving filtrate that has not been completely absorbed by the water absorbent body 7. The vent hole 14 is for discharging the air in the space 13 compressed by the filtrate, and is provided in the container body wall 10 or the container body bottom 12 so as to communicate with the atmosphere. Due to the presence of this vent 14, the entire amount of sample blood is quickly naturally filtered. It is desirable to fit a bacterial filter 16, such as a cotton plug, into the vent hole 14 to prevent the intrusion of germs. When the vent hole 14 is provided in the bottom part 12 of the container body,
A barrier 17 is provided around the vent 14 to prevent the stored filtrate from flowing out from the vent 14. It is also desirable to provide concentric barriers throughout the bottom 12 of the container body to prevent filtrate from collecting around the vent 14.

The blood bacteria isolation and culture device having the above configuration improves the reliability of the test by performing sterilization treatment using gamma ray irradiation or ethylene oxide gas.

The shape of the separation culture vessel constructed in this way is not particularly limited, but it is generally desirable to have a circular shape.

The size of the container is not particularly limited, but for example, when using two samples of blood as a specimen, the diameter of the container body must be approximately 60 mm to accommodate this. , it is better to keep it to a certain extent.

The blood sample thus treated with the blood processing device of the present invention is poured onto the filter 6 of the separation culture device described above, and the open opening is covered with the lid 9. Blood is naturally filtered by its own weight.

Bacteria in the blood remain on the filter 6, blood passes through the filter 6 and is absorbed by the water absorbent 7, and blood in excess of the saturated amount drips and accumulates at the bottom 12 of the container body. Since the air compressed by the filtrate is discharged from the vent 14, filtration is smoothly performed by the weight of the sample blood. After the filtration is completed, bacterial colonies are formed on the filter 6 by maintaining the separation incubator at a constant temperature. Bacteria are collected from the colony and subjected to bacterial tests such as bacterial identification 1 and drug susceptibility testing.

In addition, when the water absorbent body 7 is impregnated with a liquid medium and dried,
In blood processing operations, it is not necessary to add a liquid medium.

(2) Specific Effects of the Invention According to the blood processing device of the present invention, it is possible to hemolyze and coagulate blood in a sealed state. Therefore, it is possible to prevent environmental contamination due to blood bacteria, and 1) it is possible to prevent contamination of external bacteria into the specimen, thereby increasing the reliability of the test.

Furthermore, by using the blood processing device of the present invention, hemolysis, anticoagulation, and mixing operations with culture media can be simplified, and environmental contamination caused by test bacteria caused by transplantation into an incubator after treatment can be reduced. Contamination with bacteria is also minimized.

Furthermore, by using the blood processing device of the present invention, all the I-homococcal bacteria in the collected sample blood are isolated and cultured as they are, so the detection rate of the 7-homococcal bacteria is high, and the number of bacteria in the sample can be measured. It is also possible.

[Brief explanation of drawings]

FIG. 1 is a 1-meter cross-sectional view showing one embodiment of the blood processing device of the present invention, and FIG. 2 is a cross-sectional view of another embodiment. FIG. 3 is a sectional view of a blood bacteria isolation and culturing device used in a blood bacteria isolation and culturing method together with the blood processing device of the present invention. 1... Container, 2... Mixed solution of hemolytic agent and anticoagulant, 3... Column stopper, 4... Cylindrical body, 5... Pusher, 6
... Filter, 7 ... Water absorbing body, 8 ... Container body, 9
...Lid, 14...Vent

Claims (4)

[Claims] (1) A blood processing device that contains a solution containing at least a hemolytic agent and an anticoagulant, has a blood storage space, and is sterilized. (2) The blood processing device according to claim 1, wherein the association solution contains →≠medium. (3) The blood processing device according to claim 1 or 2, wherein the blood storage space is under reduced pressure. (4) The blood processing device according to claim 1 or 2, wherein the container comprises a cylindrical body and a pusher arranged so as to slide on the inner wall of the cylindrical body in a fluid-tight manner.