JPH034168A - Method and device for deciding flocculationability of blood platelet and means for deciding flocculationability - Google Patents

Abstract

PURPOSE:To objectively decide flocculationability with high accuracy and high reliability by determining the flocculation rate at a common reference point of time from plural kinds of time-flocculation rate curves and determining the pattern of concn. of blood platelet flocculation inducing agent-flocculation rare curves therefrom. CONSTITUTION:The blood platelet flocculation inducing agent is added in concn. of, for example, 3 stages concentrations to the multiplatelet plasma separated from blood and the flocculation rate curve with lapse of time is formed by a time-flocculation rate curve forming means 20. The flocculation rates of the common reference point of time after the end of the reaction with the platelet flocculation inducing agent are respectively determined from plural kinds of the time-flocculation rate curves. These flocculation rates are plotted with respect to the concn. of the platelet flocculation inducing agent to form 21 the flocculation inducing agent concn.-flocculation rate curve. The flocculationability is decided on the bases of such pattern. The flocculationability is measured objectively with the high accuracy and high reliability in this way.

Description

Detailed Description of the Invention (Industrial Field of Application) The present invention provides a method for adding a platelet aggregation-inducing agent to platelet-rich plasma separated from blood for the onset of thrombotic diseases, treatment and prevention of arteriosclerosis, etc. The present invention relates to a method and apparatus for determining platelet aggregation ability from the respective aggregation states by adding the platelet at varying concentrations in multiple stages, and an aggregation determining device.

[Conventional technology]

“Pharmacology and Treatment” (Life Science Publishing Co., Ltd., V.
ol, 1, No. 2, Feb, 1985.481~
(page 498) contains a paper on platelet aggregometry and clinical applications. That is, the curve pattern of the aggregation rate over time of platelet-rich plasma to which a platelet aggregation-inducing agent has been added is typically classified into one category as shown in FIG. In other words, TYPEI is first-order agglomeration, TYPE2 is 1
Dissociation is slow with only secondary aggregation, TYPE 3 has lower secondary aggregation than primary aggregation, TYPE 4 has higher secondary aggregation than primary aggregation, TYPE 5 has a high degree of aggregation, and the boundary between secondary and secondary aggregation cannot be distinguished. This is the case. In sp^ without additives, spontaneous aggregation appears in about 1 to 5 minutes.

When using platelet inhibitors to treat obstructive cerebrovascular disorders, myocardial embolism, etc., platelet aggregation-inducing agents are added to subjects at varying concentrations in multiple stages, and one of the patterns corresponds to each. or decide whether to use the primary or secondary
By checking the following maximum east vehicle, the degree of acceleration, normality, or suppression was determined, and the effectiveness of treatment was determined and monitored.

(Problem to be Solved by the Invention) However, when based on the classification of the pattern of the time-aggregation rate curve or the aggregation rate level, it is not possible to directly correlate with platelet aggregation, and therefore it is difficult to make a judgment based on experience. However, the results tend to be subjective.

Therefore, an object of the present invention is to provide a method, an apparatus, and a determination tool that enable objective determination of platelet aggregation ability.

(Means for Solving the Problems) As a method for achieving this object, the present invention provides multiple time-aggregation rate curves over time of platelet-rich plasma separated from blood and added with a platelet aggregation-inducing agent. The aggregation rates at a common reference point after the end of the reaction to the platelet aggregation-inducing agent are determined from these multiple types of aggregation rate curves, and these aggregation rates are calculated for platelet aggregation-inducing agent concentrations at each stage. An aggregation-inducing agent concentration-coagulation II rate curve is created by plotting against the aggregation-inducing agent concentration, and the aggregation ability is determined based on the pattern.

At this time, in order to improve the judgment accuracy, calculate the area of the aggregation-inducing agent concentration-aggregation rate curve and the time to aggregation rate at the reference time-aggregation rate curve, and calculate the area value of each area value. It is also conceivable to create it by plotting it against concentration.

In order to quickly and accurately determine the pattern of the aggregation-inducing agent concentration-aggregation rate curve, the horizontal axis is the platelet aggregation-inducing agent concentration and the vertical axis is the aggregation rate. It is convenient to use a ruler or recording paper for determining platelet aggregation ability on which a typical aggregation-inducing agent concentration-aggregation rate curve is drawn.

As a device for automatically creating an aggregation-inducing agent concentration-1ji aggregation rate vehicle according to the method of the present invention, platelet-rich plasma separated from blood and added with a platelet aggregation-inducing agent is transmitted through light while stirring, and the light is received. a photoelectric platelet aggregation measurement means for measuring a concentration from the amount; an aggregation rate curve creation means for creating a time-aggregation rate curve for platelet-rich plasma over time from a detection signal of the measurement means; Regarding the aggregation rate curve data for multiple concentration levels of the agent, the aggregation rates at a common reference point after the end of the reaction to the platelet aggregation-inducing agent are determined, and these aggregation rates are plotted against the concentration of the platelet aggregation-inducing agent to calculate aggregation. An aggregation-inducing agent concentration-aggregation rate curve creation means for creating an aggregation-inducing agent concentration-aggregation rate curve, and an output device for displaying or printing out the aggregation-inducing agent concentration-aggregation rate curve created by the curve creation means. to be accomplished.

(Function) Platelet-rich plasma that has been separated from blood, added with a platelet aggregation-inducing agent, and stirred undergoes an aggregation phenomenon corresponding to the characteristics of the additive over the course of one hour. Therefore, by changing the concentration of the aggregation-inducing agent in multiple stages, a plurality of time-aggregation rate curves in which the aggregation rate is plotted sequentially over time can be obtained.

Based on this curve data, when the aggregation rate value at the reference time point when the reaction is completed is plotted against the change in the concentration of the coagulation 4J inducing agent, an inducing agent concentration-aggregation rate curve is obtained. Therefore, based on this curve data, it is possible to determine in what pattern the aggregation rate changes with respect to the change in the aggregation concentration. In addition, it has been experimentally confirmed that the concentration at which secondary aIN begins to appear (on the dye collection side 1 is a concentration corresponding to about 50% of the maximum aggregation rate), and therefore the agglutination threshold can be immediately determined. , it is possible to immediately determine whether aggregation is normal or not, and the degree of enhancement or suppression.In this determination, it may be determined by applying a ruler for platelet aggregation ability determination or by plotting on a platelet aggregation ability determination recording paper. can.

According to the device of the present invention, stiffness! The inducing agent concentration-aggregation rate curve is automatically created, and the area of the aggregation rate curve up to the reference point aggregation rate is calculated, and the aggregation inducing agent concentration-aggregation rate curve as each area value data is also automatically created. You can also create one.

〔Example〕

An example of the means for measuring platelet aggregation of the present invention will be described.

A platelet aggregation-inducing agent such as ADP (^denos 1ne) is added to platelet-rich plasma (PRP) separated from the collected blood.
Di-Phosphate), for example, in 3 stages 1. c+μ
M (mole), added at a concentration of 2.0 μm, 4.0 μm,
As shown in FIG. 1, the aggregation rate curve of PRP over time is measured. At this time, platelets perform primary a collection by adding ADP to PRP, and when platelets are activated by 80P stimulation, a release reaction occurs and endogenous ADP
New secondary aggregation occurs, and both primary and secondary aggregation can be observed with an appropriate amount of ADP.

In other words, ^DP1.0, the aggregation caused by H is a primary aggregation-dissociation type in which the aggregates are almost completely dissociated;2. Aggregation due to ΩμM results in biphasic aggregation! 2. Primary and secondary Raito type agglomeration, in which dissociation of agglomerates does not occur, and agglomeration due to 4.0 pot test are non-dissociated type aggregation, in which aggregation occurs following initial agglomeration, and is secondary aggregation that cannot be distinguished from primary agglomeration. It exhibits aggregation due to endogenous ADP.

For these three types of time-aggregation rate curves, calculate the aggregation rate at a reference point, for example, 5 minutes after the start of measurement, when the various reactions described above have finished and almost reached an equilibrium state, and As shown in FIG. 2, the values are plotted against ^DR11 degrees to create an inducing agent concentration-aggregation rate curve. Then, from this curve pattern, a normal state is immediately determined by comparing it with a standard curve that has been experimentally created in advance.

It was also found that the maximum aggregation rate was approximately 80%,
The agglomeration threshold therefore corresponds to 40% of its height.1.
It turns out that it is about 7p. It should be noted that if the number of concentration steps is further increased, a correspondingly more variety of time-aggregation rate curves can be obtained, and a layer-precise elicitor concentration-aggregation rate curve can be obtained.

When making such a determination, if a transparent ruler 1 shown in FIG. 3 is used, it can be immediately determined to which type the aggregation-inducing agent concentration-aggregation rate curve plotted above belongs or to which aggregation threshold value it belongs. This ruler has been used by many healthy people, hyperintensity patients,
For example, standard aggregation-inducing agent concentration-aggregation rate curves prepared by the method of the present invention for 8 levels of A-DP for hyperactive patients, mildly hyperactive patients, and suppressed patients are plotted on the horizontal axis ^DP
The concentration and the aggregation rate are plotted on the vertical axis, and the classification is shown in the table below. It should be noted that even if the aggregation inducing agent is the same, the curve may deviate depending on the solution, etc., and the one-dot line indicates the range of variation among the types to which it belongs.

That is, when this is used, the standard curve shown in FIG. 3 overlaps the plot point in FIG. 2 shown by the x mark, and IIJ is immediately determined, and at the same time, the aggregation threshold value is also found on the scale of the ruler. In addition, such a plot can be plotted at the standard aggregation inducing agent concentration in Figure 3.
Even when performed on recording paper for platelet aggregation ability determination on which an aggregation rate curve is printed, the determination is similarly accurate and immediate. Furthermore, the data indicated by the O mark in Fig. 3 similarly indicates that ADP is 0.0.
, 5.1.0, 1.5, 2,0, 2.5, ko, 0. 4.0 w extinction (Q, This is a case where the ruler l is applied to the data of 8 levels of concentration.

It is more clearly determined to be type m.

Depending on the type of aggregation-inducing agent, it may be necessary to create a standard pattern that is not based on the 5-minute value but at another reference point such as the 3-minute value or the 10-minute value.

FIG. 4 shows the configuration of an example of a platelet aggregation ability determining apparatus for carrying out the method of the present invention.

In the figure, reference numeral 10 denotes a photoelectric aggregation measurement means, and as is well known, Cubela-11, which collects PRP separated from blood and added with a platelet aggregation-inducing agent, is set in a heat-retaining tank 14. Stir with Stara 12.

It is configured to irradiate light from one side with a light source 13 and make the transmitted light enter the light receiving element 13a.

As is well known, the light receiving element 13a receives the PRP from the star 1.
It is adjusted to output a detection signal at a level corresponding to a translucent aggregation rate value of 0% when set to 2 and 100% when platelet-poor plasma (ppp) is set.

20 is a time-aggregation rate curve creating means, and the light receiving element 13
Based on the detection signal of a, an aggregation rate curve of PRP over time is created. Reference numeral 21 denotes an aggregation-inducing agent concentration-aggregation rate curve creation means, which creates a common reference point at which time has elapsed for significant aggregation to occur corresponding to the characteristics of the aggregation-inducing agent for the time-aggregation rate curves at multiple levels of concentration. The aggregation rates of each are determined, and from these aggregation rate data, an aggregation rate curve is created in which the aggregation rate is plotted against the aggregation-inducing agent concentration. 22 stores a standard pattern shown in FIG. 2 in the memory 22a, and compares this standard pattern with the measured aggregation-inducing agent concentration-aggregation rate curve to determine which type or aggregation threshold value it is. This is a curve creation means. These circuit means 20, 21, 22 can be composed of a CPU. 23 is a printer that records the aggregation-inducing agent concentration-aggregation rate curve created by the circuit means 22 and 23, together with the specified type or agglutination threshold of this curve pattern.

With such a device, the cuvette 11 is reset each time the concentration is changed in one port of the aggregation rate measurement means, and photoelectric measurements are performed on multiple types of PRP, so that the time-aggregation rate curve creation means 20 can measure multiple types of PRP each time. PRP of
Along with creating an agglomeration rate curve.

The aggregation-inducing agent concentration-aggregation rate curve creation means 21 creates an aggregation-inducing agent concentration-1B rate curve from the aggregation rates at each reference point, and automatically sends the test subject's platelet aggregation ability data to the printer 23. output. At this time, the recording paper of the printer may be printed with a standard pattern as shown in FIG.

If the agglutination-inducing agent concentration-actual car curve creation means 21 is configured to calculate the area of the time-to-aggregation rate curve up to the agglutination rate value at the reference time and to plot each surface area value, the reference point is calculated. An aggregation-inducing agent concentration-aggregation rate curve can be easily created based on the area value rather than the aggregation rate value itself at the time, and discrimination between determination curves can be made more clearly.

(Effects of the Invention) As described above, according to the present invention, 'r4 determination is efficiently performed in order to form a single meaningful curve for the data to be determined, and from that pattern, platelet aggregation ability can be determined objectively. ·High precision·
Judgment is made under high reliability. Further, the aggregation threshold value, which is the concentration of the aggregation-inducing agent at which secondary aggregation begins to occur, can be easily estimated by reading the concentration at a certain ratio of the aggregation rate to the maximum actual vehicle.

The determination will be extremely efficient and accurate if a ruler or recording paper for determining platelet aggregation ability is used that depicts standard aggregation-inducing agent concentration-aggregation rate curves for inhibited, normal, and enhanced aggregation.

According to the apparatus of the present invention, an aggregation-inducing agent concentration-aggregation rate curve is automatically created.

[Brief explanation of the drawing]

Figure 1 is a diagram showing the time-aggregation rate curve of platelet-rich plasma according to an example of the method for determining platelet aggregation ability of the present invention, and Figure 2 is a diagram showing the 5-minute value of aggregation rate versus R concentration according to the same example. Figure 3 showing the aggregation-inducing agent concentration-aggregation rate curve plotted.
The figure shows a platelet aggregation ability determination ruler used for determining the aggregation-inducing agent concentration-aggregation rate curve according to the same example, and FIG. FIG. 5 is a diagram showing the configuration of the 2-East fusion foot device and a diagram showing a typical aggregation rate curve over time of platelet-rich plasma to explain the conventional determination method. l...judgment ruler

Claims (1)

[Claims] 1) Create time-aggregation rate curves over time for platelet-rich plasma separated from blood and added with a platelet aggregation-inducing agent for multiple levels of platelet aggregation-inducing agent concentrations; From each type of time-aggregation rate curve, the aggregation rate at a common reference point after the end of the reaction to the platelet aggregation-inducing agent is determined, and these aggregation rates are plotted against the platelet aggregation-inducing agent concentration to determine the aggregation-inducing agent concentration. - A method for determining platelet aggregation ability, which comprises creating an aggregation rate curve and determining aggregation ability based on the pattern of the aggregation-inducing agent concentration-aggregation rate curve. 2) An aggregation-inducing agent concentration-aggregation rate curve is created by calculating the area of the time-to-aggregation rate curve at the reference time point against the platelet aggregation-inducing agent concentration and plotting the respective area values. The method for determining platelet aggregation ability according to claim 1. 3) A platelet aggregation ability determination ruler for determining the pattern of the aggregation-inducing agent concentration-aggregation rate curve in the platelet aggregation ability determination method according to claim 1 or 2, wherein the ruler is set on a transparent plate with the horizontal axis representing platelet aggregation. A ruler for evaluating platelet aggregation ability, characterized in that the aggregation-inducing agent concentration and the aggregation rate are plotted on the vertical axis, and standard aggregation-inducing agent concentration-aggregation rate curves are drawn for inhibition, normality, and enhancement of aggregation, respectively. 4) A platelet aggregation ability determination recording paper for determining a pattern of an aggregation-inducing agent concentration-aggregation rate curve in the platelet aggregation ability determination method according to claim 1 or 2, wherein the aggregation-inducing agent concentration-aggregation rate curve is Standard aggregation-inducing agent concentration-aggregation rate curves for suppression, normality, and promotion of aggregation are printed on printed or handwritten paper, with the horizontal axis representing platelet aggregation-inducing agent concentration and the vertical axis representing aggregation rate. A recording paper for determining platelet aggregation ability, characterized by the following. 5) A photoelectric platelet aggregation measuring means for measuring the degree of aggregation based on the amount of light received by transmitting light while stirring platelet-rich plasma separated from blood and added with a platelet aggregation-inducing agent; an aggregation rate curve creation means for creating a time-aggregation rate curve with respect to the passage of time of platelet-rich plasma from a detection signal; an aggregation-inducing agent concentration-aggregation rate curve creating means for calculating aggregation rates at a common reference point and plotting these aggregation rates against platelet aggregation-inducing agent concentration to create an aggregation-inducing agent concentration-aggregation rate curve; and an output device for displaying or printing out the aggregation-inducing agent concentration-aggregation rate curve created by the curve creation means. 6) The aggregation-inducing agent concentration-aggregation rate curve creation means calculates the time to the aggregation rate at the reference time with respect to the platelet aggregation-inducing agent concentration.
6. The platelet aggregation ability determination device according to claim 5, wherein the area of the aggregation rate curve is calculated and the respective area values are plotted to create an aggregation-inducing agent concentration-aggregation rate curve.