JPS6058555A - Blood coagulation measuring method and apparatus therefor - Google Patents

Abstract

PURPOSE:To obtain the titled apparatus capable of performing two kinds of measurements by one operation and the predetermined blood coagulation measurement rapidly and accurately, by a method wherein a liquid mixture of serum and a coagulating reagent is irradiated with light to collect scattered light or transmitted light which is, in turn, converted to an electric signal and the change amounts of a time and the signal when said signal reaches a predetemined level are calculated. CONSTITUTION:In measuring a prothrombin time and a fibrinogen concn., a calcium added tissure thromboplastin reagent is warmed at first and serum is added to said reagent while the resulting mixture is introduced into a thermostatic reaction cell (a). At this time, a controller 5 receives a start signal. Monochromatic light is incident to the cell (a) from LED of a light emitter (e) and the scattered light thereof is received by the photodiode of a photoelectric converter (d) and converted to an electric signal having intensity corresponding to the degree of blood coagulation. This analogue signal is amplified and converted to a digital signal by an A/D converter (e) to be inputted to a microcomputer MiC. This MiC receives the start signal from the switch 4 of a pipet 2 and transmits an output signal to a motor 11 to rotor 10 at a high speed and the serum in the cell (a) is mixed with the coagulating reagent.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application This invention relates to a blood coagulation measuring method and a blood coagulation measuring apparatus.

Structure of conventional examples and their problems Conventionally, bleeding time and whole blood coagulation time tests have been performed as tests for bleeding disorders, but recently prothrombin 1lfrlli (
FT), partial thromboplastin time (PTT), activated partial thromboplastin time (APTT), and fibrinogen concentration (Fbg ing.

Inspection costs are also high.

The method for measuring each of the above items is outlined below.

PT: 3 ml of calcium-added 51m thromboplastin reagent
Pre-wet the tube at 7°C, add plasma to it, and measure the total time until it coagulates.

P′rT t APTT ): A mixture of partial thromboplastin reagent (activated partial thromboplastin reagent) and plasma is heated at 37°C for a certain period of time, and a pre-filled calcium chloride solution is added to it. Measure the time until solidification.

Fbg: Fibrinogen standard solution (fibrinogen concentration known) diluted in Oronperonal buffer solution (diluted in several steps) Prepare all. Add thrombin reagent to each diluted fibrinogen standard solution and measure the time until coagulation. Draw a calibration curve for the fibrinogen concentration and the obtained coagulation time on a logarithmic graph. For a 10-fold dilution of the test plasma with an unknown concentration, add the thrombin reagent and similarly calculate the coagulation 1i/l!l. The fibrinogen concentration is determined by measuring the fibrinogen concentration.

As mentioned above, in the measurement of fibrinogen concentration (Fbg), operations such as plasma dilution and preparation of a calibration curve are complicated;
Measurement takes time. In addition, the reagents used are expensive, and the overall cost of the test is quite high.Blood coagulation tests are often performed in all medical cases where bleeding is expected, and tests must be performed as quickly as possible. However, the current situation is that it has not been possible to meet the above-mentioned demand for a rapid test because it has conventionally taken a long time to measure the fibrinogen concentration.

In view of the actual circumstances beyond the purpose of the invention, the first invention is to provide a method for quickly performing blood coagulation measurements, and the second invention is to provide an apparatus that can quickly and accurately perform blood coagulation measurements. The blood coagulation measuring method of the first invention, which has the overall purpose of achieving the following, consists of: (1) a process of completely mixing a blood inkstone and a coagulation reagent, and (2) irradiating the mixed liquid with a source to detect scattered light or transmitted light. The prothrombin time (PT) is calculated from the process of capturing the electrical signal and converting it into an electrical signal, and the time taken for the electrical signal to reach a predetermined level and the amount of change in the electrical signal.
or a process of determining partial thromboplastin time (PTT) or activated partial thromboplastin time (APTT) and fibrinogen concentration (Fbg).

The reagent for measuring FT and Fbg in one operation is the reagent for PT measurement described above, and the reagent for measuring PTT and Fbg in one operation is the reagent for PTT measurement 11 described above, The same applies to the case where APTT and Fbg are measured in one operation. Fbg is F T ,
It has been experimentally confirmed that substantially the same value is obtained when measuring both PTT and APTT.

According to the method of the first invention, light and electrical signals are used to immediately convert PT, PTT, or APT from a change in the electrical signal.
Since T and Fbg can be measured and two types of measurements can be performed in one operation, it is possible to significantly shorten the time compared to conventional methods.

Further, as shown in FIG. 1, the blood coagulation measuring device of the second invention has: (1) a cell for injecting plasma and a coagulation reagent, and (2) a content liquid k? ? ? , means for stirring; ■ a light emitter C that irradiates light to the cell a; and ■ a photoelectric converter d that receives scattered light or transmitted light from the cell a and converts it into an electrical signal. , ■ Δ/1) converter C that converts the electrical signal into a digital signal, and a microcomputer MiC, for example, is used. ■1 The value of the digital signal from the A/D converter e is expressed as (2) a means f2 for determining whether the change in the digital signal has stopped; and (3) the time from the start of inputting the digital signal until reaching a predetermined level in the digital signal change area (i.e., FT or i- j：
Means f3 and possible 4 for calculating PTT or APTTJ
means f4 for calculating the total amount of change in the digital signal and converting it into a value corresponding to fibrinogen concentration (Fbg);
and all of them, and further 0 each of the arithmetic means f3. f, and means g for displaying the values obtained.

Description of Examples The constant temperature reaction SEFVA in which plasma and coagulation reagents are injected is placed in a constant temperature bath of 1 yen as shown in Figure 2.
1) R solid reagent is added by pressing the push button 3 of the pipette 2 before sorting, and the plasma and coagulation reagent are instantly mixed by stirring means HLC provided outside the thermostatic chamber 1.

The Bibev l-211c is provided with a switch 4 that recommends VC when the push button 3 is pressed. On the other hand, the stirring means is composed of a rotating body 10 made of an elastic material such as rubber and a motor 11 having a shaft that fixes the rotary body in a completely eccentric state. Motor 1
When 1 is rotated at high speed, the lower part of cell a rotates at high speed,
A stirring effect similar to that obtained by shaking can be obtained.

Examples of the light emitter C include a light emitting diode and an incandescent lamp. A laser or the like is used. As the photoelectric converter d, for example, a photodiode, a photofungistor, a CdS
Yuko etc. are used. The photoelectric converter d is provided at a position where only the scattered light from S/L/a enters, while the transmitted light from S/L/a enters.

・1 after 12 interfaces with A/D converter ef
The microcomputer MiC connected to the controller 5,
Arithmetic unit 6. A numeric display 8 and a printer 9 are used as a 0 display means g which includes a memory 7, a, and a generation means i. The microcomputer MiC sends the VC setting of the temperature control sub-circuit 14 of the heater 13 of the thermostatic chamber 1. It also sends an on signal to the motor control sub-circuit and an off signal '(il-) after the timer time has elapsed.

We will explain the dynamics of prothrombin time (FT) and fibrinogen concentration (Fbg) when all J is constant.

First, a sample of 77 μm of force/L/1-romboblastin was prewarmed at 37° C., and plasma 2 was added to it and placed in a thermoplastic reaction chamber. This time control unit 5 receives a start signal indicating 0 points.

Cell alc monochromatic light is input from the light emitter C, that is, the light emitting diode (LED J, and the scattered light is received by the electric converter d, that is, the photodiode VC, and the intensity of the scattered light is determined by blood and V coagulation (suspension). 1 of strength according to the degree of
converted into a difference signal. This '1H, 1 word is an analog signal, and after being amplified by an amplifier, it is
The signal is converted into a digital signal by the converter eVcj and inputted to the microcomputer MiC.

The microcomputer MiC is the swiss of Vibe J+-2.
Upon receiving the start signal from the edge 4, the timer in the device is started, and at the same time, the motor IIK output signal is transmitted for a certain short period of time to rotate the rotating body 1° at high speed. By pressing, the plasma in the cell/L/a and the coagulation reagent are mixed instantly.

The controller 5 has a clog generating means i (clovk generator) which is started by the start signal, and works in cooperation with the memory device 7 and the computing device 6 to generate the clotting generator from the time when the plasma and the coagulation reagent are mixed. Printer 9' (i
1- drive to print the prototype. An example of 10-bit printing is shown in Figure 'ti3.

The output does not change at a certain level until point A, where the reaction progresses and fibrin is about to be formed, and then the scattered light suddenly increases, passes through point B, and reaches point C, where the increase stops. This change is actually quite drastic.

The middle height of the waist is 40% if point A is 0% and point 6 is 100%.
point B)
T).

A value calibrated by multiplying the voltage output difference HTh between points A and C in digital quantity by a certain coefficient is used as the fibrinogen concentration (Fbg), and on the numerical display 8, the above 7'' al-thrombin time ( PT) Displayed together with T.

The functional flowchart for the microcomputer MiC described above is shown in FIG. 4, and the execution program flowchart is shown in FIG. 5, which will be explained below.

Initialize with stay 77'' (11, stay, t 7-(21
Wait for the start signal from the switch 4 of the float 2, and if there is an input, start the black puffer at step (31). Then, input the sample clock at step (41).・Increase the count by +1f.Step (61 reads data VIII from the A/D converter e, VIII is the input voltage value in the clock count IK.In step +71, transfer the data to VtI+ and the gross count It memory. do.

All drive signals are transmitted to the display means g using the stainless steel knob (8).

In the Ste knob (91), the deviation of the input voltage value V tl
+ -V (I-1) = Δ■ is calculated, and step-lip (
10), it is determined whether the deviation ΔV is 0 or not. Then, it is determined whether the 0 point has been reached in Figure 3+Al.

If NO in this judgment, return to step (4) and read, display, and store the data again. If YES, proceed to step (111) and check whether the vv mountain is equal to or greater than the specified value 70.
In other words, in the prisoner in Figure 3, perform all discrimination as to whether it is before point A or reached point 0. If NO, return to Stennob (4) and select YES.
If so, that is, if it reaches 0 points, stay knob (12)
Move to.

Stay 77” (121VC oil, final f[VIN+
The deviation H from the initial value V(11) is calculated, and step (13
) with deviation HIC calibration gold line 9. You can calculate the value by multiplying it by a certain coefficient. This value Hs is fibrinogen me (
In step (14), the fibrinogen concentration [1-1sk is 0, which is displayed on the numerical display 8, and then the time tm corresponding to 4096 of the deviation H is calculated.

First, step 9 (151T operation Vllll + (40/1
001 X H= Vm'e, then stay J]"
(16), the two closest values V between Vm'i and nlC
Read (K1. v (K-) 1 ) from the table in memory, the third [1911B+
Calculation of proportional allocation of Δ of (Vm - V[KI) / (
V (KI1) - VIKI] = Δ, S T
, t 7'' (18) IC O, prothrombin time T is calculated by K(K+Δ)Xto=T.Here, t
o is the period of the clorif. At the stainless knob (19), the prothrombin time T is displayed on the numerical display 81C. Then, the data obtained in the last step (2o) is printed.

In addition, 40% of 10 thrombin time (P"l")'tH
The time taken is based on experience.

However, in the third (2) tAuc, the rise from point A to point c is actually quite steep, and as a practical matter, 1
Even if the time is equivalent to 0% to 90%, there is not much error.

In addition, partial thromboplastin 11J1 (P'FT
) and activated partial thrombosis: 'lJ? ? 174
] (APTT) +7) It was experimentally confirmed that the fibrinogen concentration (Fbg) @Hs (=K-H) is almost equal to the value in the prothrombin time (PT) measurement. The rise from point A to point C was similarly steep.

Fig. 6 shows an example of the correlation between fibrinogen concentration and voltage deviation H of prothrombin time (Fig. 3 in PTI measurement) by the conventional method, and Fig. 7 shows the correlation between fibrinogen concentration and activated partial thromboplastin time (Fig. 3 in PTI measurement) by the conventional method. As an example of the correlation with the voltage deviation H in the APTT) measurement, a follow-up test of m confirmed that the H value has sufficient validity as a fibrinogen concentration value. The equation of the regression line in Figure 6 is y = 0.23 x + 18.95 and the correlation coefficient is r = 0.855, and the equation of the regression line in Figure 7 is 7 = 0.19 x + 6.88. The correlation coefficient is r = 0.901. In addition, as shown by the line in Figure 2, the photoelectric converter d can also receive the transmitted light from the cell a; in this case, the The graph corresponding to Figure 3 is a graph that continues at a constant high level, then falls sharply, and then reaches a constant low level.

Effects of the Invention According to the blood coagulation measuring method of the first invention, prothrombin time (PT) or partial thromboplastin time (PT
r ) or activated partial thromboplastin time (AP
TT) and fibrinogen concentration (Fbg).In other words, it has the advantage of being able to quickly perform two types of measurements with just a single cI operation. Further, according to the blood coagulation measuring device of the second invention, there is an effect that the measurement value can be determined extremely accurately using the measurement method according to the first invention.

[Brief explanation of drawings]

Fig. 1 is a block diagram showing the configuration of the second invention, Fig. 2 is a block diagram of the embodiment, Fig. 3 +A+ is a time/output correlation graph, Fig. 3 [Bl is Fig. 3 + point B of AI] Enlarged view of nearby area,
Figure 4 is a functional flowchart, Figure 5 is a flowchart of the execution program, Figure 6 is a correlation graph between H and fibrinogen concentration during prothrombin time measurement, and Figure 7 is H and fibrinogen concentration during activated partial thromboplastin time measurement. This is a correlation graph. a... Cell, b... Stirring means, C... Light emitter, d
...Photoelectric converter, e...A/D converter, fl...
Storage means, f2...Signal change stop determination means, f3. f
4...Arithmetic means, g...Display means No. 3 Figure 4 Figure 4 - Q-t-0, S'IJ4 animals are resistant to WE, but Keyaki exchange meeting ヱO\enemy To 1 section 10he Ichiinka C-Ko Z Hill

Claims (1)

[Claims] (11) A process of completely mixing plasma and a coagulation reagent, a process of irradiating the mixed liquid with light and sound to capture scattered light or transmitted light, and converting it into an electrical signal, A method for measuring blood coagulation that includes the process of calculating the prothrombin time or partial thromboplastin time or activated partial thromboplastin time and fibrinogen concentration from the time when a predetermined level is reached and the amount of change in the electrical signal. (21 Plasma and Coagulation) A cell for injecting the reagent, a means for stirring the 7 μm content liquid, a light emitter for irradiating the SE/I/VC light, and a device for receiving all of the scattered or transmitted light from the cell and converting it into an electrical signal. a photoelectric converter for converting the electric signal, an A/D converter for converting the electric signal into a digital signal, and the A/D converter for converting the electric signal into a digital signal;
The value of the digital signal from the converter along with the elapsed time is
means for storing E3; means for calculating the entire time period from the start of input of the digital signal until reaching a predetermined level of the digital signal change region; and means for calculating the entire change layer of the digital signal to a value corresponding to the fibrinogen concentration. 1. A blood coagulation measuring device comprising means for converting into a value, and means for displaying the values obtained by each of the arithmetic means.