JPH08105885A - Blood coagulation accelerating agent, blood coagulation accelerating method, and blood inspection container - Google Patents

Abstract

PURPOSE: To obtain a coagulation agent which does not affect coagulation activity or inspecition by allowing a carrier to carry an antifungal metal and incoporating an antifungal composition which is insoluble to blood for constituting a blood coagulation accelerating agent. CONSTITUTION: For example, zerolite, montmorillonite, and ceramic whose specific gravity is equal to or more than approximately 1.03 are used as carriers, and, for example, silver, copper, and zinc are used as antifungal metals. The carrier carries the antifungal metals as antifungal compositions by ion exchange, complexing, clathrate and the particle diameter is set to approximately 0.01-500μm. The antifungal compositions are mixed with such minerals as glass, kaolin, and bentonite, and such organic blood coagulation acceleration substance as ellagic acid for adjusting a blood coagulation accelerator, thus enabling the antifungal composition itself to maintain coagulation acceleration activity, without reducing entire specific activity and unfavorable influence to a blood inspection value is negligible.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the field of clinical examination for examining a blood sample, and more particularly, to shorten the blood coagulation time in the step of coagulating a blood sample and separating serum components by centrifugation or the like. The present invention relates to a blood coagulation promoting agent, a blood coagulation promoting method, and a blood test container therefor.

[0002]

2. Description of the Related Art In recent years, with advances in testing technology, blood tests such as chemical tests, immunoserologic tests, and hematological tests have become remarkably mechanized, and as long as suitable pretreated samples can be prepared. Inspection is possible in a short time, for example,
The outpatient can now obtain a doctor's diagnosis based on the test results on the spot when he / she visits the hospital, which greatly contributes to speeding up the diagnosis of diseases.

Of these, the pretreatment in the hematological examination is
Sufficient mixing of the anticoagulant and blood is sufficient, does not require much time, and can be immediately set in the measuring device. However, in chemical tests or immunoserologic tests, in order to obtain the necessary serum, it is necessary to once coagulate the blood and then to separate the serum components by centrifugation etc. It costs. Therefore, in order to shorten the time required for all steps of the test from pretreatment of the sample to obtaining the measurement result, it is not enough to mechanize the analysis step, and the fractionation time of serum is also shortened. There must be.

By the way, the blood used for the test is stored,
Conventionally, a glass container has been widely used as a blood test container used for coagulation and separating serum by centrifugation. However, this is vulnerable to mechanical shock, and if it is damaged, there is a risk of infection due to spilled or scattered blood samples, and there is also the problem of increased burden on the patient due to re-collection of blood. The percentage of containers is increasing.

However, the plastic container is not practical because it has a poor activation power for blood coagulation factor XII, factor XI and the like, and as it is, it takes a much longer time for blood coagulation as compared with glass products. Therefore, as disclosed in Japanese Patent Application Laid-Open No. 58-195151, mineral substances such as glass, kaolin, bentonite, silica, and celite; fine powder of a substance having a blood coagulation accelerating property such as ellagic acid is preliminarily treated with blood. It is applied to the inner wall surface of an inspection container, or as disclosed in JP-A-58-105064, a carrier such as a nonwoven fabric or a plastic sheet which is substantially insoluble in blood and chemically inert. It is attempted to shorten the blood coagulation time by, for example, accommodating the one carrying the above-mentioned fine powder in a container.

As described above, when a substance having a blood coagulation accelerating property is applied to the inner wall surface of a blood test container or supported on a carrier, first, these fine powders are added to purified water or alcohol / purified water mixed medium. Prepare a suspended treatment solution and spray this treatment solution on the inner wall surface, or dip a carrier in this treatment solution and dry it. Cut it into an appropriate shape and store it in a blood test container. Things to do are being done.

However, this treatment liquid is usually defenseless against microbial contamination, and if handled improperly, there is a risk of microbial contamination. Furthermore, when the treatment liquid contains a water-soluble polymer compound such as polyvinylpyrrolidone, which is usually used as a binder of fine powder of a coagulation promoting substance or as a viscosity modifier of the treatment liquid, modified cellulose, Since the water-soluble polymer compound (1) serves as a suitable nutrient source for microorganisms, the risk of microbial contamination of this treatment liquid is further increased.

Condensate may be generated in the treatment solution as the growth of microorganisms progresses, the spray nozzle may be clogged, the uniformity of the coated surface may be significantly impaired, and the carrier may be immersed in the treatment step. Problems such as unevenness in the density of loading on the carrier and measurement errors are likely to occur. The risk of such microbial contamination is not limited to the alteration itself of this treatment liquid, and if the method of storing the blood test container manufactured using this treatment liquid is inappropriate, it will always be present even during storage. Is.

Therefore, a blood test container containing a carrier carrying a coagulation-promoting substance on its inner wall surface, unless it is manufactured in a special aseptic environment, is exposed to radiation such as γ-rays or electron beams, or It is necessary to sterilize using a chemically reactive gas such as ethylene oxide gas. In any of these sterilization methods, the sterilization conditions such as radiation dose, reactive gas concentration, heating temperature, and exposure time must be adjusted depending on the contamination status before sterilization, which is extremely complicated. Operations are required.

Further, when the sterilized object is heavily contaminated, the object to be sterilized may be irreversibly denatured or deformed due to severe sterilization conditions. Furthermore, even in the storage after sterilization, if the proper packaging state that blocks the invasion of microorganisms is not maintained, the sterilization of the corners becomes useless.

One of the effective means for solving these problems is to impart antibacterial properties to the coagulation promoter itself. As a result, the above-mentioned microbial contamination can be suppressed, and even if sterilization becomes necessary, the conditions can be alleviated, and the physical and chemical denaturation that the sterilization brings to the object to be sterilized is suppressed. can do. Further, the packaging form of the blood test container can be simplified.

By the way, as the antibacterial and antifungal agents generally used for preventing microbial contamination, many compounds have been known and widely used for food additives. However, most of these antibacterial and antifungal agents are water-soluble, so if blood is collected in a blood test container made from a suspension of a blood coagulation promoter containing them, the antibacterial and antifungal agents are contained in the blood. The mold agent may melt out and adversely affect various chemical tests. Further, when the antibacterial and antifungal agent is composed of a water-soluble heavy metal salt, it also denatures the enzyme relating to blood coagulation and eliminates the enzyme activity, which inhibits blood coagulation and causes the separation of serum, etc. It may be difficult to achieve the purpose of the term.

In view of the above, the present invention has a blood coagulation promoting property which does not substantially affect blood coagulation activity and serum chemistry tests, and which itself contains an antibacterial composition having a blood coagulation promoting action. The purpose is to provide an agent.

[0014]

SUMMARY OF THE INVENTION The gist of the present invention is to provide an antibacterial composition, which comprises a carrier carrying an antibacterial metal and is substantially insoluble in blood when the blood coagulation promoter is constituted. It exists in the place to contain. The gist of the present invention is also a blood coagulation promoting method comprising contacting the blood coagulation promoting agent with blood, and further, a blood test container containing the blood coagulation promoting agent.

The carrier used in the present invention is required to be removed from the serum during the centrifugation for separating the serum after blood coagulation, and the specific gravity of human serum is 1.02. The specific gravity of the carrier is preferably 1.03 or more, and more preferably 1.05.
That is all.

The carrier is not particularly limited in addition to the specific gravity described above, and examples thereof include inorganic materials such as zeolite, montmorillonite, ceramics, glass and sparingly soluble phosphate; organic materials such as graphite and ion exchange resins. Among them, zeolite, montmorillonite, ceramics, silicic acid compounds such as sparingly soluble phosphate, and sparingly soluble phosphoric acid compounds themselves have a blood coagulation accelerating property, and are therefore very suitably used.

The antibacterial metal used in the present invention is not particularly limited, and examples thereof include copper and silver of Group Ib of the periodic table; II.
Group b zinc, cadmium, mercury; group IVa germanium, tin, lead; metal salts such as lanthanoid cerium; and organometallic compounds, among which silver, copper, zinc,
Cerium is preferably used because it has a good balance between toxicity and usefulness.

The antibacterial composition of the present invention comprises the above carrier carrying the above antibacterial metal. Examples of the structure for supporting the antibacterial metal on the carrier include ion exchange, complexation and inclusion. Other than these, it is not preferable because it may be eluted in the blood.

The above-mentioned antibacterial composition is preferably in the form of fine powder having a large specific surface area and has a particle size of 0.01-.
500 μm is preferred. When the particle size is less than 0.01 μm, the antibacterial effect is enhanced, but after blood coagulation is completed, even if centrifugation is performed to separate serum, 1000 to 18
Under normal centrifugation conditions of about 00G for 5 minutes, it easily remains in the serum, causing turbidity of the serum, and the carried metal is measured redundantly with the metal originally remaining in the serum. Therefore, problems such as giving a positive error to the inspection are likely to occur, which is not preferable.

If the particle size exceeds 500 μm, the antibacterial composition is apt to be in a non-uniform dispersed state when a suspension of the blood coagulation promoter is prepared, and the antibacterial property is exhibited. Since it is localized in the vicinity of the surface of, the antibacterial and antifungal effects of the entire suspension cannot be expected, which is not preferable.
More preferably, it has an average particle diameter of 0.01 to 50 μm.

The minimum amount of the above-mentioned antibacterial composition to be used should be determined based on MBC (minimum bactericidal concentration against bacteria) and MFC (minimum bactericidal concentration against fungi) as in general antiseptics and fungicides. However, it is possible to determine the composition so that the concentration of the antibacterial agent in the suspension of the blood coagulation promoter according to the present invention, for example, a suspension using purified water, is 0.1 μg / ml or more. preferable. The maximum amount of the above antibacterial composition to be used is such that when a large amount of an insoluble substance is suspended in blood, undesirable side effects such as hemolysis may occur. Therefore, the concentration when mixed in blood is 0.5 μg / ml or less. It is preferable to determine the compounding composition so that

The blood coagulation promoting agent of the present invention comprises the above antibacterial composition, a mineral substance such as glass, kaolin, bentonite, silica and celite; and an organic blood coagulation promoting substance such as ellagic acid. Can be mixed and prepared using various methods.

The above-mentioned blood coagulation promoter is suspended in purified water or physiological saline to prepare a suspension, which is added to the collected blood and mixed to bring it into contact with blood to coagulate blood. The time can be shortened.

The above blood coagulation promoter is suspended in purified water, alcohol / purified water mixed medium or the like to prepare a suspension, which is sprayed on the inner wall surface of the blood test container, or
After immersing a carrier such as a nonwoven fabric or a plastic sheet in the suspension, the dried product is cut into an appropriate shape,
By accommodating in a container or the like, a blood test container having a high blood coagulation accelerating property can be prepared.

The above-mentioned suspension of blood coagulation promoter may also be used as a binder of fine powder of coagulation-promoting substance or as a viscosity modifier of this suspension, a water-soluble polymer compound such as polyvinylpyrrolidone or modified cellulose. May be included.

[0026]

The antibacterial composition containing silver, copper, zinc and the like, zeolite, montmorillonite, ceramics and sparingly soluble phosphate contained in the blood coagulation promoter of the present invention is purified by suspending the promoter. It not only kills microorganisms invading water, alcohol / purified water mixed medium, etc., but also effectively controls microbial contamination during storage of blood test containers made by using this promoter, and also this antibacterial Since the sexual composition does not dissolve in blood, it does not impair the function of the blood coagulation promoting substance and does not adversely affect the blood test value. Furthermore, since the antibacterial composition itself also has blood coagulation accelerating properties, the specific activity of the blood coagulation accelerating agent as a whole is not reduced. The above-mentioned antibacterial metal contained in the antibacterial composition of the present invention has an antibacterial property even though it hardly elutes in a free ion state because active oxygen is generated in the vicinity of the supported metal. Conceivable.

[0027]

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

Examples 1 to 12 and Comparative Example 1 Confirmation of antibacterial activity Purified water suspensions of blood coagulation promoters were prepared according to the composition shown in Table 1. In Table 1, Bactekiller is manufactured by Kanebo.
BM103A, ice is NAZ, manufactured by Catalyst Kasei Kogyo Co., Ltd.
320, Rasp is AN600 manufactured by Rasa Kogyo Co., Ltd.
Novalon represents AG300 manufactured by Toagosei Co., Ltd., respectively.

On the other hand, separately from this, silica (1.0
%) And polyvinylpyrrolidone (2.0%) to prepare a purified water suspension, which was exposed to indoor air to cause microbial contamination, and this was used as a suspension of inoculum for inoculation. . With respect to each of the suspensions according to Table 1, the preparation as it is, and the mixture that was stirred for 10 hours after forcibly causing microbial contamination by inoculation with inoculum were used,
Bacterial and fungal culture tests were performed according to standard practice. The culture medium and culture conditions are as follows:
A Girl (Standard Method Aga)
r) was used for culturing at 30 ° C. for 3 days, the inoculum of each suspension was 0.3 ml, and for fungi, potato dextrose agar (Potato Dextrose) was used.
Agar) was cultivated at 25 ° C. for 3 days, and the inoculation amount of each suspension was 0.3 ml, and a culture test was conducted.
The relative frequency of the colonies formed in the medium was measured, and the results in the bacterial medium and the fungal medium are shown in Table 2 and Table 3, respectively. In each table, − means that no colony was generated, and +++ means that colonies were extremely generated.

Examples 13 to 20 Confirmation of Effect on Blood Coagulation Purified water suspensions of blood coagulation promoters were prepared according to the compositions shown in Table 4. Approximately 50 μl of each suspension was sprayed onto a polymethylmethacrylate blood collection tube having a volume of 10 ml, and dried with a blast dryer at 60 ° C. Blood collection tube 1
About 3 ml of fresh rabbit blood was collected per book and left standing at room temperature of 23 to 25 ° C. The time at which blood loses fluidity and starts to exude was measured as blood coagulation time. The results are shown in Table 5.

Examples 21 to 28 Confirmation of effects on blood test values After the evaluations described in Examples 13 to 20 were completed, each was subsequently centrifuged at 1800 G for 5 minutes,
Serum was collected from each sample and representative biochemical test items (GOT and ALT as enzymes, TG, PL, and T-as lipids) were collected.
CHO, electrolytes such as Na, K, Cl, Mg, Ca, and metal components such as Fe and Cu) were investigated. The results are shown in Table 6. In Table 6, the blood collection tubes of Examples 21 to 28 correspond to the blood collection tubes of Examples 13 to 20, respectively.

Comparative Examples 2 to 4 Purified water suspensions of blood coagulation promoter containing no antibacterial composition were prepared according to the compositions shown in Table 4. About 50 μl of this suspension was sprayed on a 10 ml-volume blood collection tube made of polymethylmethacrylate and dried by a blow dryer at 60 ° C. (Comparative Example 2). Separately from this, a 10 ml capacity hard glass blood collection tube (Comparative Example 3) and a polymethylmethacrylate blood collection tube (Comparative Example 4) not sprayed with a blood coagulation promoter were prepared. Example 1 for these three types of blood collection tubes
In the same manner as in 3, fresh rabbit blood was collected and blood coagulation time was evaluated. The results are shown in Table 5 together with the results of Examples 13 to 20.

Comparative Examples 5 to 7 After the evaluation described in Comparative Examples 2 to 4 was completed, each of them was subsequently centrifuged at 1800 G for 5 minutes, and serum was collected in the same manner as in Example 21. The effect on biochemical test values was investigated. The results are shown in Table 6 together with Examples 21 to 28. In Table 6, the blood collection tubes of Comparative Examples 5 to 7 correspond to the blood collection tubes of Comparative Examples 2 to 4, respectively.

As is clear from the results of Tables 2 and 3,
Immediately after preparation, when a microorganism invades a suspension of a coagulation promoter free from microbial contamination, the microorganism easily grows when the antimicrobial composition is not included (Comparative Example 1). In each case (Examples 1 to 12), no colony was detected, indicating that the microorganism was killed.

From the results shown in Table 5, the polymethylmethacrylate blood collection tube (Comparative Example 4) containing no coagulation-promoting substance (comparative example 4) required one hour or more for blood coagulation. When the coagulation accelerator according to (Examples 13 to 20) is used, coagulation is completed in 20 to 30 minutes,
Results that are substantially comparable to the results obtained with the conventional coagulation-promoting component that does not contain an antibacterial composition (Comparative Example 2), and are comparable to those obtained with a hard glass blood collection tube (Comparative Example 3). Therefore, it was found that the antibacterial composition of the present invention not only does not adversely affect blood coagulation but also has positive coagulation promoting properties. From the results of Table 6, it was found that none of the antibacterial compositions had a substantial adverse effect on the blood test.

[0036]

[Table 1]

[0037]

[Table 2]

[0038]

[Table 3]

[0039]

[Table 4]

[0040]

[Table 5]

[0041]

[Table 6]

[0042]

The blood coagulation promoter of the present invention contains an antibacterial composition in which an antibacterial metal is supported on a carrier composed of a silicic acid compound or a sparingly soluble phosphoric acid compound, and the effect of simply mixing microorganisms is effective. Not only does not inhibit the coagulation-promoting activity of the original coagulation-promoting substance, but the antibacterial composition itself also has coagulation-promoting activity. The specific activity of the whole substance is not reduced, and the adverse effect on the blood test value can be substantially ignored, which is extremely useful.

Claims (5)

[Claims] 1. A blood coagulation promoter comprising an antibacterial metal supported on a carrier and containing an antibacterial composition substantially insoluble in blood. 2. The blood coagulation promoter according to claim 1, wherein the carrier is a silicic acid compound or a sparingly soluble phosphoric acid compound. 3. The blood according to claim 1 or 2, wherein the antibacterial metal is at least one metal ion selected from the group consisting of silver, copper, zinc, cadmium, mercury, germanium, tin, lead and cerium. Coagulation accelerator. 4. A method for promoting blood coagulation, which comprises contacting the blood coagulation promoter according to claim 1, 2 or 3 with blood. 5. A blood test container containing the blood coagulation accelerator according to claim 1.