JPH06337267A - Dry reagent for measuring blood coagulation time - Google Patents

Abstract

(57) [Summary] (Modified) [Structure] Tissue thromboplastin, calcium salts such as calcium chloride, bovine adsorbed plasma, magnetic particles such as ferric tetroxide particles, sugars such as glucose, and polyoxyethylene sorbitan monooleate. Blood coagulation containing at least one additive selected from the group consisting of surfactants such as glutamic acid, amino acids such as glutamic acid, amino acid salts such as sodium glutamate, proteins such as BSA, and polyhydric alcohols such as PEG6000 Timed dry reagent. [Effect] According to the present invention, the solubility of the TT dry reagent for thrombotest (TT) measurement is improved, and the simultaneous reproducibility is remarkably increased. Therefore, it becomes possible to provide a TT dry reagent suitable for practical use, and when the anticoagulant is administered by surgery or the like, it is possible to quickly monitor the coagulation ability of the patient's blood.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a blood coagulation time measuring dry reagent for measuring thrombotest activity used in clinical tests.

[0002]

2. Description of the Related Art The thrombotest (hereinafter abbreviated as TT) measurement is a blood coagulation test item provided for the purpose of monitoring the effect of oral anticoagulant administration.

[0003] Since oral anticoagulants inhibit the action of vitamin K in vivo, the administration thereof reduces the activities of blood coagulation factors II, VII, X and the like which are activated by the action of vitamin K. Therefore, the effect of oral anticoagulant administration is
Since it becomes clear by examining the activity of these coagulation factors, a method of performing evaluation by performing a blood coagulation assay that reflects these activity of the factors is generally used.

When TT is measured, the activity of blood coagulation factors II, VII and X can be comprehensively judged from the measurement results. The feature of the measurement with the TT reagent is that blood coagulation factors I and V are added to the reagent in advance. Blood coagulation factors I and V have an effect on the coagulation time, which is unrelated to the original activity value, when monitoring the effect of oral anticoagulant administration. However, the TT measurement is most suitable for monitoring because the measured value can be stably obtained by adding these factors to the reagent in advance.

By the way, all of the conventionally used TT measurement methods use a solution-type reagent and are still widely used. This method is a method in which a reagent in the form of a solution is first prepared, a blood sample is added thereto and reacted, and a change in turbidity or a change in viscosity of the reaction solution is monitored to determine coagulation. The example will be specifically described below.

The TT reagent contains tissue thromboplastin, bovine adsorbed plasma, and calcium salt, and a lyophilized product is generally commercially available. The dried product TT Reagent T in the form of an aqueous solution
Prepare a T reagent solution and preheat it. next,
Similarly, the pre-heated plasma or whole blood is mixed with the TT reagent solution, the change in turbidity or the change in viscosity is monitored immediately after addition, and the number of seconds until coagulation is measured to determine the coagulation time. The coagulation time obtained as described above is the coagulation time of TT, which is prolonged as the coagulation factor activity decreases with the administration of the oral anticoagulant.

On the other hand, a simple blood coagulation assay system using a dry reagent has recently been developed (see Table 3
No. 504076). The principle of the assay system is
A dried reagent containing magnetic particles in a blood coagulation reagent is prepared, a blood sample is added to the dried reagent, and immediately after that, the magnetic particles are moved by applying a combination of an oscillating magnetic field and a static magnetic field. The motion signal is optically monitored to detect coagulation.

The above publication shows a number of assay methods using the system and applying various blood coagulation reagents in dry reagents. However, the above publication makes no mention of a blood coagulation reagent in the dry reagent as a TT reagent, that is, a dry reagent for TT measurement and a specific description of TT measurement using the dry reagent.

[0009]

In the TT measuring method by the solution method which is currently used, an operation such as preparation of a reagent solution is carried out for the purpose of the TT measuring of rapid and accurate monitoring of the effect of oral anticoagulant administration. However, there were major problems such as being complicated and time consuming, and being able to do this only in a place equipped with containers and measuring instruments. Further, there is a problem that the liquid reagent has poor storability and it is not economical unless the whole amount is used. However, it was considered that the above-mentioned problems would be solved by the method using a dry reagent.

Therefore, according to the method for producing a dried reagent for blood coagulation assay described in the above-mentioned publication, TT
A dry reagent for measurement was manufactured as follows. That is, TT
Magnetic particles were added to the reagent solution to give a suspension, which was dispensed on a reaction slide and freeze-dried. When plasma was added dropwise to this dried reagent and an attempt was made to measure the coagulation time, there were major problems that the reproducibility of the coagulation time was poor and the measurable activity value range was narrow, and it was unusable at all. It was Therefore, it has been found that a dry reagent for TT measurement that can withstand use cannot be produced by simply using the method for producing a dry reagent for measurement items other than TT described in the above publication.

[0011] The problem to be solved by the present invention is to provide a practical reagent for blood coagulation time measurement (hereinafter also referred to as TT dry reagent) for TT measurement which can be put to practical use, and T using the same.
Development of T measurement method.

[0012]

[Means for Solving the Problems] As a result of intensive studies to solve the above problems, it was found that the improvement of reproducibility of coagulation time of TT dry reagent and the expansion of measurable activity value range were observed in blood samples. It has been found that this can be achieved by improving the solubility of the TT dry reagent when added.

Further, it has been clarified that the solubility of the reagent can be improved by preparing a TT dry reagent by adding a specific additive, and further research has led to the present invention. .

That is, the present invention relates to tissue thromboplastin,
Drying blood coagulation record containing calcium salt, bovine adsorbed plasma, magnetic particles, and at least one additive selected from the group consisting of saccharides, surfactants, amino acids, amino acid salts, proteins, and polyhydric alcohols. It relates to reagents.

The method for preparing the TT dry reagent of the present invention is not particularly limited, but a method in which an aqueous solution containing the above essential components is prepared as a final solution for TT dry reagent, aliquots are dispensed on a reaction slide, and freeze-dried is used. It is common. The details will be described below.

The material of the reaction slide for dispensing the final solution for TT dry reagent is to optically monitor the increase in the viscosity of the reaction solution consisting of the blood sample and TT dry reagent as attenuation of the motion signal of magnetic particles during TT measurement. There is no particular limitation as long as it can be used, and typically, polyester, polyvinyl chloride, or the like is used.

As an example, there is a reaction slide as shown in FIGS. FIG. 1 is a view of the reaction slide as viewed from above. The part surrounded by the dotted line in FIG. 1 is the main part consisting of the final solution dispensing port for TT dry reagent and the plasma addition port. FIG. 2 shows the structure of the main part in detail.
A main part is constituted by laminating a transparent polyester plate B on a normally white polyester plate C, and then laminating a transparent polyester plate A on B.

When the above-mentioned final solution for TT dry reagent is put in through the dispensing port shown in FIG. 1, the portion D is filled with the final solution. Then, the TT dried reagent of the present invention can be prepared by retaining the reagent components in the reaction slide by freeze-drying or air-drying.

The contents of the respective constituents in the TT dry reagent of the present invention may be arbitrarily determined, and representative numerical values are shown below. The content of each component described below is the content when 20 μl of the final solution for TT dry reagent was dispensed to a reaction slide and dried.

Tissue thromboplastin as used in the present invention is a blood coagulation factor extracted from bovine brain, which is a component that initiates coagulation of a blood sample. Tissue thromboplastin is generally used in an amount of 2 × 10 ⁻⁸ to 2 × 10 ⁻³ g, but 2 × 10 ⁻⁶ g to 2 × 1 from the viewpoint of more stable progress of the coagulation reaction.
^0-4 g is preferred.

The calcium salt is a supply source of calcium ions essential for the coagulation reaction, and specific examples thereof include calcium chloride and calcium lactate. Generally, the calcium salt is used in an amount of 1.1 × 10 ⁻⁶ g to 2.2 × 10 ⁻⁴ g, but 2.2 × 1 in terms of more stable progress of the coagulation reaction.
0 ⁻⁶ g to 1.1 × 10 ⁻⁵ g is suitable.

Bovine adsorbed plasma is prepared by removing blood cells from bovine blood by centrifugation or the like and further removing blood coagulation factors II, VII, IX, and X, and is a source of blood coagulation factors I and V. Therefore, it may be powder or liquid. As a method for removing blood coagulation factors II, VII, IX, and X, for example, a usual method of adsorbing and removing with barium sulfate powder or the like can be used. Generally, bovine adsorbed plasma is 8 × 10 ^-5 g ~
As used 1.5 × 10 ^-3 g ^{but, 2 × 10 -4 g~6 × 10} -4 g from the viewpoint of progression of the coagulation reaction becomes more stable are preferred.

The magnetic particles are used to monitor the increase and decrease of the viscosity of the reaction solution based on their motion signals, and for example, ferromagnetic substances such as iron sesquioxide particles, iron sesquioxide particles, nickel particles and cobalt particles are preferably used. However, from the viewpoint that the intensity of the obtained signal is large and stable, iron tetroxide particles are particularly preferable. The size of the magnetic particles can be generally 10 μm or less in average particle size, but particles having an average particle size of 0.4 to 0.9 μm can be preferably used from the viewpoint that the intensity of the obtained signal is higher. The content of magnetic particles is usually 2 x
It is used at 10 ⁻⁶ g to 2 × 10 ⁻⁴ g, but 2 × 10 ⁻⁵ g to 1.2 × from the point that the obtained signal intensity is higher.
10 ^-4 g is preferred.

The additive improves the solubility of the TT dry reagent, and is selected from the group consisting of saccharides, surfactants, amino acids, amino acid salts, proteins, and polyhydric alcohols, which may be used alone or in combination of two or more. Used in combination.

Examples of the saccharides include oligosaccharides such as glucose, fructose and sucrose, and polysaccharides such as dextran. Any of them can be used, but the solubility of the TT dry reagent is better and stable. Oligosaccharides are preferably used. The saccharide is usually 2 × 10 ⁻⁵ g to 4 × 10
^Although it is used in an amount of ⁻³ g, 1 × 10 ⁻⁴ g to 1 × 10 ⁻³ g is preferable because the solubility of the TT dry reagent is better and the solubility is uniform.

Surfactants include cationic surfactants such as benzalkonium chloride and undecyltrimethylammonium bromide, anionic surfactants such as sodium cholate and sodium deoxycholate, and 3-[(3 −Cholamidop
ropyl) dimety lammonio] -1 -propanesulfonate (CHA
PS), 3-[(3-Cholamidopropyl) dimetylammonio] -2
-Both ionic surfactants such as hydroxy-1-propanesulfonate (CHAPSO) and nonionic surfactants such as polyoxyethylene alkyl ether can be used, but the solubility of TT dry reagent is better and stable Therefore, a nonionic surfactant can be preferably used. The surfactant is usually used in an amount of 2 × 10 ⁻⁵ g to 2 × 10 ⁻³ g, but 1 × 10 ⁻⁴ g to 1 × in terms of better solubility and uniformity of the TT dry reagent. 10 ^-3 g is preferred.

Examples of amino acids or salts thereof include neutral amino acids or salts thereof such as glycine, alanine and glycine hydrochloride, acidic amino acids or salts thereof such as glutamic acid, aspartic acid and sodium glutamate, and basic amino acids or salts thereof. Examples include lysine, lysine hydrochloride, arginine, etc., which can be used,
An acidic amino acid and an acidic amino acid salt can be preferably used because the TT dry reagent has better solubility and stability. The amino acid or salt thereof is usually 2 × 10 ⁻⁵ g to 2 × 1.
Although it is used at 0 ^-3 g, the solubility of the TT dry reagent is better and uniform, so that it is 1 x 10 ^-4 g to 1 x 10 ^-3 g.
Is preferred.

As the protein, for example, casein, gelatin, bovine serum albumin (hereinafter abbreviated as BSA) and the like can be used. The protein is usually 2 × 10 ⁻⁵ g to 2 ×
Although it is used at 10 ^-3 g, the solubility of the TT dry reagent is better and the solubility is 1 × 10 ^-4 g to 1 × 10 ^-3.
g is preferred.

Examples of the polyhydric alcohol include glycerol and polyethylene glycol (hereinafter abbreviated as PEG), and any of them can be used. However, PEG is preferable because of its better solubility and stability of the TT dry reagent. It can be preferably used, and particularly has an average molecular weight of 5,000 to 10,000.
Those are preferable. The polyhydric alcohol is usually 2 × 10 ^-5
g-2 × 10 ^-3 g, but 1 × 10 ^-4 g-1 because the solubility of the TT dry reagent is better and uniform
× 10 ^-3 g is preferable.

Among these additives, sugars and polyhydric alcohols are preferable because they have particularly stable solubility and good reproducibility of coagulation time. When the above additives of the present invention are not used at all, the solubility of the TT dry reagent is remarkably lowered, and reproducible coagulation time cannot be obtained, so that TT measurement is difficult.

In the dry reagent for measuring blood coagulation time according to the present invention, at least one additive selected from the group consisting of tissue thromboplastin, calcium salt, bovine adsorbed plasma, ferromagnetic magnetic particles, and sugars and polyhydric alcohols. The contained blood coagulation time measurement dry reagent is excellent in solubility between the blood sample and the reagent, and as a result, the motion signal intensity of the obtained magnetic particles is large and determination is easy, and the measurement sensitivity is improved. Is a preferable reagent in various respects such as good reproducibility, good correlation with the conventional solution method, and the like.

The method for preparing the TT dry reagent of the present invention is not particularly limited, but a commercially available dry TT reagent containing the aforementioned tissue thromboplastin, bovine adsorbed plasma and calcium salt is used, and this dry TT reagent is diluted with water. After reconstitution, an aqueous solution of magnetic particles and an additive is added, or an aqueous solution of magnetic particles and an additive is added directly to a dried TT reagent to prepare a final solution for a TT dry reagent, and the final solution is transferred to a predetermined reaction slide. A typical preparation method is a method of dispensing a fixed amount and drying.

Water is usually used as the above-mentioned liquid for reconstitution and the liquid for dissolving the additives.
8 buffer of any concentration, such as 20 mM Tris buffer (pH 7.3), 20 mM HEPES buffer (pH 7.
3) etc. may be used.

The freezing method for preparing the TT dry reagent may be either an instant freezing method using liquid nitrogen or the like, or a slow freezing method, but the solubility of the TT dry reagent is better and uniform. Therefore, the method of freezing slowly is preferable. To exemplify the freezing method, after a predetermined amount of the final solution for TT dry reagent was dispensed on a predetermined reaction slide at room temperature,
A method of placing in a freezer at -40 ° C and freezing slowly can be used.

The drying method may be either freeze-drying or air-drying, but freeze-drying is preferred because the solubility of the TT dry reagent is more uniform. To exemplify the freeze-drying method, the temperature is changed from -30 ° C to 30 ° C in a vacuum state.
A method of increasing the temperature and freeze-drying can be used.

The measurement of TT with the TT dry reagent of the present invention is not particularly limited, but typically, a certain amount of a blood sample is dropped onto the TT dry reagent and immediately subjected to a combination of an oscillating magnetic field and a static magnetic field to form magnetic particles. The coagulation can be detected by allowing the magnetic particles contained in the TT dry reagent to be optically moved and at the same time being optically monitored. Such measurement is carried out under the trade name of CO
AG1 [Wako Pure Chemical Industries, Ltd.] and trade name CG-01
[(A & T Sales Co., Ltd.]] can be used.

When the TT is measured by the above method, the T
T coagulation time refers to the time from the addition of a blood sample to the coagulation point. The freezing point can be determined based on the change in the intensity of the motion signal of the magnetic particles in the TT dry reagent. As the freezing point, the point where the decay rate of the signal intensity is maximum,
Alternatively, a point at which the signal intensity is attenuated to an arbitrary ratio with respect to the maximum value is used. Since the reproducibility of the obtained coagulation time is better, the point where the decay rate of the signal intensity is maximum can be preferably used.

When evaluating the effect of oral anticoagulant administration, T
Sometimes the T activity value is used. The TT activity value is used for standardization to eliminate the difference in TT coagulation time between models.

As a method for obtaining the TT activity value of an arbitrary blood sample using the TT dry reagent of the present invention, a general method of calculating from the obtained TT coagulation time using a standard curve can be used. The method of preparing the standard curve is not particularly limited, but for example, a blood sample of known TT activity value is arbitrarily diluted in 5 steps, and the vertical axis of the log-log graph is the coagulation time and the horizontal axis is T.
A method of creating the T activity value can be preferably used.

As a result of the improved reagent solubility according to the present invention,
For the first time, a TT dry reagent suitable for practical use for TT measurement can be provided. Therefore, it is most suitable for urgent TT measurement.
A measurement method that does not require preparation of a reagent solution has become possible.

[0041]

【Example】

Example 1 Reproducibility of TT Dry Reagent Containing Additive The TT dry reagent was produced as follows. Complex factor T Kokusai [International Reagents Co., Ltd.] which is a commercially available TT reagent;
Including a bovine brain-derived tissue thromboplastin, bovine adsorbed plasma and calcium lactate] according to the instruction manual to prepare an aqueous solution, replace the water with a glucose aqueous solution prepared to have a final glucose concentration of 1% by weight. Was added. Further, Fe ₃ O ₄ particles (manufactured by Rare Metallic Co., Ltd.) were added so as to have a final concentration of 5 mg / ml to prepare a final solution for TT dry reagent, which was shaken and stirred well. 20 μl of the final solution was dispensed to each reaction slide shown in FIG.
It was frozen in a freezer at 40 ° C. and then freeze-dried. Freeze-drying was performed by a method of linearly increasing the temperature from −30 ° C. to 20 ° C. in 7 hours in a vacuum state.

The TT measurement using the TT dry reagent was carried out as follows. Cytolol I used as a sample
(Normal model plasma, manufactured by Dade Baxter) was prepared as instructed in the instruction manual, and then equal amount of Orenberonal buffer (manufactured by Sigma) was added to dilute to 1/2 concentration and TT dried. 25 μl was used for measurement for each reagent.
The TT dried reagent stored at 4 ° C after freeze-drying was allowed to stand in a desiccator until it reached room temperature, then CG01 [Co.
A & T Sales] was used to measure the coagulation time described below twice. When the movement of magnetic particles after addition of the sample is measured as a change in scattered light, the intensity of this signal sharply decreases as the viscosity increases with coagulation. Immediately after adding the sample,
As shown in Table 1, the inflection point at which the decay rate of the signal intensity is the maximum is the freezing point, and the freezing time is from the addition of the sample to the freezing point. The results are shown in Table 1. The maximum signal intensity is also shown in Table 1. The higher the solubility of the TT dry reagent, the higher the maximum signal intensity.

From Table 1, T containing the above-mentioned glucose was added.
The T-dry reagent has good reproducibility when measured twice.
It turns out that it is suitable for use in measurement. Further, it can be seen that the maximum signal intensity is large and the reagent is sufficiently dissolved.

Example 2 Reproducibility of TT Dry Reagent Containing Additive In the same manner as in Example 1, a non-ionic surfactant polyoxyethylene was used instead of the glucose aqueous solution when the final solution for TT dry reagent was prepared. A TT dry reagent was prepared by adding an aqueous solution of sorbitan monooleate. TT measurement with the prepared TT dry reagent was also performed in the same manner as in Example 1,
The results are shown in Table 1.

From Table 1, it can be seen that the above-mentioned TT dry reagent containing polyoxyethylene sorbitan monooleate has good reproducibility when measured twice and is suitable for use in TT measurement. Further, it can be seen that the maximum signal intensity is large and the reagent is sufficiently dissolved.

Example 3 Reproducibility of TT Dry Reagent Containing Additive In the same manner as in Example 1, a sodium glutamate aqueous solution was added in place of the glucose aqueous solution at the time of preparation of the final solution for the TT dry reagent to prepare a TT dry reagent. I went. TT measurement using the prepared TT dry reagent was also performed in the same manner as in Example 1, and the results are shown in Table 1.

Table 1 shows that the above-mentioned TT dry reagent containing sodium glutamate has good reproducibility when measured twice and is suitable for use in TT measurement. Further, it can be seen that the maximum signal intensity is large and the reagent is sufficiently dissolved.

Example 4 Reproducibility of TT Dry Reagent Containing Additive In the same manner as in Example 1, a BSA aqueous solution was added in place of the glucose aqueous solution when the final solution for TT dry reagent was prepared,
A TT dry reagent was produced. TT measurement with the prepared TT dry reagent was also performed in the same manner as in Example 1, and the results are shown in Table 1.
It was shown to.

From Table 1, it is understood that the above-mentioned TT dry reagent containing BSA has good reproducibility when measured twice and is suitable for use in TT measurement. Further, it can be seen that the maximum signal intensity is large and the reagent is sufficiently dissolved.

Example 5 Reproducibility of TT Dry Reagent Containing Additive In the same manner as in Example 1, an aqueous solution of PEG6000 (manufactured by Wako Pure Chemical Industries; average molecular weight 7500) was used instead of the glucose aqueous solution when the final solution for TT dry reagent was prepared. Was added to produce a TT dry reagent. TT by the prepared TT dry reagent
The measurement was performed in the same manner as in Example 1, and the results are shown in Table 1.

From Table 1, it can be seen that the above-mentioned TT dry reagent containing PEG6000 has the best reproducibility when measured twice and is suitable for use in TT measurement. Further, it can be seen that the maximum signal intensity is large and the reagent is sufficiently dissolved.

Comparative Example 1 Reproducibility of TT dry reagent containing no additive An TT dry reagent was prepared in the same manner as in Example 1, except that no additive was added at the time of preparing the final solution for TT dry reagent. . TT measurement using the prepared TT dry reagent was also carried out in Example 1.
The results are shown in Table 1.

From Table 1, it can be seen that T without addition of any of the above additives.
It can be seen that the T-dried reagent is remarkably inferior in reproducibility when measured twice. Also, it can be seen that the maximum signal intensity is very small and the reagent is hardly dissolved. From the above results, when the TT dry reagent was prepared without using the additive of the present invention, it was difficult to use it for the measurement of TT, and the coagulation that can be used for the measurement of TT only after using the additive. It can be seen that it shows time reproducibility.

[0054]

[Table 1]

Example 6 TT activity standard curve A TT dry reagent was prepared in the same manner as in Example 1 using PEG6000 as an additive. As a measurement sample, human standard plasma [International Reagents Co., Ltd., Thrombotest activity 110%] was used as a diluted solution to prepare diluted plasma having TT activity of 11, 22, 55, and 82.5%, and the coagulation time of each TT activity was measured. Diluent is B
SA 5%, sucrose 10%, sodium chloride 0.9
An aqueous solution containing 100% was used. The measurement results are shown in FIG.

From the measurement results, a good standard curve was obtained on the log-log graph as shown in FIG. Therefore, when the reagent of the present invention is used, the standard curve is calculated to obtain T from the coagulation time.
It can be seen that the T activity value can be calculated.

Example 7 Factor Susceptibility Test A TT dry reagent was prepared in the same manner as in Example 1 using PEG6000 as an additive, and the blood coagulation factor X sensitivity was examined. The measurement samples were prepared using normal plasma and blood coagulation factor X-deficient plasma (George King). Normal plasma has a factor content of 100%, and factor-deficient plasma has a factor content of 0.
Two types of plasma were mixed and used so that the factor content was 5, 10, 20, 50, or 75%. The measurement results are shown in Figure 5.
It was shown to.

From FIG. 5, it can be seen that the reagent of the present invention has sufficient sensitivity to blood coagulation factor X.

Example 8 Correlation with Conventional Method Using PEG6000 as an additive, a TT dry reagent was produced in the same manner as in Example 1, and 38 samples of human plasma were used as measurement samples to examine the correlation with the conventional method. . The conventional measurement is T
Thrombotest Oren (Eisai) as T reagent,
KC-10 (Amelunc) was used as a measuring device according to each instruction manual. The measurement results are shown in FIG.

The relationship between the measurement method using the TT dry reagent for TT measurement of the present invention and the conventional method is Y = 17.4 + from the measurement results.
It can be expressed as 1.45X, and the correlation coefficient at this time was 0.9903. Therefore, it is understood that the TT measurement method using the TT dry reagent of the present invention has a very good correlation with the conventional TT measurement method.

Example 9 Correlation with Conventional Method Using a sucrose aqueous solution prepared to have a final concentration of 10% by weight in place of the glucose aqueous solution, a TT dry reagent was produced in the same manner as in Example 1, Human plasma 2 as a measurement sample
The correlation with the conventional method was examined using 8 samples. For the measurement by the conventional method, Thrombotest Oren (Eisai) was used as the TT reagent, and KC-10 (Amelunc Co.) was used as the measuring device according to the respective instruction manuals. The measurement results are shown in Figure 7.
It was shown to.

From the measurement results, the relation between the measurement method using the TT dry reagent for TT measurement of the present invention and the conventional method is Y = −7.95.
4 + 1.653X, and the correlation coefficient at this time was 0.9900. Therefore, it is understood that the TT measurement method using the TT dry reagent of the present invention has a very good correlation with the conventional TT measurement method.

[0063]

According to the present invention, the solubility of the TT dry reagent for thrombotest measurement when a blood sample is dropped is improved. As a result, simultaneous reproducibility of the coagulation time is improved, the motion signal intensity of the obtained magnetic particles is increased, the determination is facilitated, the measurement sensitivity is improved, and further, the correlation with the conventional solution method is good. From this point of view, an excellent TT dry reagent can be obtained.

Therefore, it becomes possible to provide a TT dry reagent suitable for practical use, and when the anticoagulant is administered by surgery or the like, it is possible to rapidly monitor the coagulability of the patient's blood.

[Brief description of drawings]

FIG. 1 is a schematic view of an example of a reaction slide for TT dry reagent.

FIG. 2 is a configuration diagram of a main part of an example of a reaction slide for TT dry reagent.

FIG. 3 is a change in signal intensity when a blood sample is added to a TT dry reagent.

FIG. 4 is a standard curve of TT activity in Example 6, where the vertical axis (logarithm) is the coagulation time and the horizontal axis (logarithm) is the TT activity value.

FIG. 5 is a graph of blood coagulation factor X sensitivity in Example 7, wherein the vertical axis (logarithm) is the coagulation time and the horizontal axis (logarithm) is the factor content rate.

FIG. 6 is a correlation graph between the present method and the conventional method in Example 8, in which the vertical axis represents the coagulation time of the present method and the horizontal axis represents the coagulation time of the conventional method.

FIG. 7 is a correlation graph between the present method and the conventional method in Example 9, in which the vertical axis represents the coagulation time of the present method and the horizontal axis represents the coagulation time of the conventional method.

Claims (1)

[Claims] 1. Tissue thromboplastin, calcium salt, bovine adsorbed plasma, magnetic particles, saccharides, surfactants,
A blood coagulation time measuring dry reagent containing at least one additive selected from the group consisting of amino acids, amino acid salts, proteins, and polyhydric alcohols.