JPS5928253B2 - Method for detecting coagulation state of blood, etc. - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for optically and electrically detecting the state of an agglutination reaction in blood or the like.

Traditionally, medical tests have been carried out to determine the physiological state of a sample by mixing reagents or serum, etc. with certain components of the sample, causing an agglutination reaction, and checking whether or not agglutination occurs in the maternal sample. ing.

Since both the operation and final judgment of these tests are performed manually, there is a considerable risk of erroneous judgments.
F. The present invention is configured to electronically perform the operation of determining the result of an agglutination reaction as either agglutination or non-aggregation,
The purpose of this invention is to provide a method that allows quick determination and highly reliable determination. When a sample subjected to an agglutination reaction on a transparent substrate is viewed optically, the following characteristics appear.

1) In the non-agglomerated state, the light transmittance in the reaction zone has a substantially uniform value with little local variation.

92) In the agglomerated state, agglomerates with low light transmittance and liquid parts with high light transmittance are generated on the reaction zone, so that the light transmittance varies greatly depending on the location.

The present invention was invented focusing on the relative characteristics of (1) and (2) above. That is, the five reaction regions are scanned with a light beam having a minute cross-sectional area to obtain the amount of light transmitted through the minute cross-sectional area, and this is photoelectrically converted into an electric signal, and this electric signal is replaced according to the above-mentioned characteristics. This system is configured to identify the agglutination state of the specimen by identifying the specimen. 0 The invention will now be explained with reference to the drawings which show an apparatus configured to carry out the method of the invention.

FIG. 1 schematically shows the detection section of this device, in which a transparent substrate 1
A reaction zone 2 is formed on top by placing a specimen and injecting a reagent.

The illustrated example shows a case where aggregation occurs. A light beam 4 having a minute cross-sectional area emitted from the light source 3 passes through the reaction area 2, is focused by the lens system 5, and enters the photoconductive element 6. The light beam 4 is moved by a combination of movements in the right angle directions as shown by arrows 7 and 8, and scans the entire reaction area 2 at a constant speed. An electric closed circuit 9 including the photoconductive element 6 is connected in series with a DC power source 10 and a fixed resistor 11. The voltage across the fixed resistor 11 becomes an electrical signal 12 having the illustrated polarity with respect to the ground potential. In this device, for example, when the light beam 4 hits an agglomerate, the amount of light transmitted is small, and therefore the amount of light irradiated to the photoconductive element 6 is also small, so that the resistance of the element 6 becomes large.

Therefore, the current flowing through the closed circuit 9 is small, and the electrical signal 12, which is the output voltage, is also small. However, when the light beam 4 is in the liquid part outside the aggregate, the amount of light transmitted is large and the photoconductive element 6 is irradiated with light, so the electric signal 12 has a large value.
In this way, the electrical signal 12 is obtained as an electrical signal having a magnitude corresponding to the amount of light transmitted through the reaction area 2. FIG. 2 is a block diagram of an identification device that uses the electric signal 12 as an input signal to determine the presence or absence of aggregates.

13 is a peak value hold circuit that holds the maximum value of the electrical signal 12 input during a certain period and outputs it as an output signal 32;
4 is a subtraction circuit that outputs a value obtained by subtracting a reference value 15 (described later) from the signal 32, and 16 is a subtraction circuit 14 that compares the output signal 33 of the subtraction circuit 14 with the electric signal 12.
This is a comparator that generates a pulse signal 34 with a constant amplitude when the signal 33 is larger than the electric signal 12 (comparison condition).

17 is a differentiator that differentiates the pulse signal 34 of the comparator 16;
A rectifier 18 passes only a negative signal from the output signal 35 of the differentiator 17, and this negative signal resets the operation of the peak value hold circuit 13.

The peak value hold circuit 13 is also reset by a constant scan completion signal 19 issued from a scan drive system (not shown) each time one scan is completed. 20 is an integrator that integrates the output signal 34 of the comparator 16 over the entire scanning time of scanning the entire reaction area, and 21 is a comparator that compares the output signal 36 of the integrator 20 with a reference value 22 (described later). If the output signal 36 of the integrator 20 is larger, a signal 23 indicating aggregation is output.

The reference value 15 is also referred to as the threshold fluctuation width,
In a reaction zone exhibiting an agglomerated state, an electric signal consisting of a voltage X large enough to indicate an agglomerate and an electric signal consisting of a voltage Y (Y>X) large enough to indicate a liquid part are generated. The potential corresponds to the potential difference (Y-X).

The reference value 22 is a scanning time corresponding to an area of an agglomerate sufficient to determine that aggregation is present, which is converted into an electrical signal.

Since each of the above reference values 15 and 22 is an important value for ensuring accuracy of determination, each is set to an appropriate size depending on the properties of the specimen and the purpose of determination.

Next, the present invention will be explained with reference to FIGS. 3 and 4 showing actual inspection examples.

Figure 3 shows the case with aggregation (column A) and the case with no aggregation (column B).
The output signal waveforms at each part of the device are shown for each column (column).

reaction zone 2a, where AO indicates aggregation and BO indicates non-agglomeration;
2b is a model diagram showing the optical characteristics of sample 2b, where AO indicates a state in which aggregates and a partially unreacted sample are both suspended in the liquid part, and BO indicates a state in which the sample is almost homogeneously distributed. . The output signal waveforms shown in A1 to A5, Bl to B4, and C are:
A among all scan lines. , one scanning line 29a shown in BO,
29b shows a waveform when scanning is performed. A1 is fixed resistance 1
1 shows an electrical signal 12a generated at both ends of 1. A2 indicates the output signal 32a of the peak value hold circuit 13. In A2, two positions C and d are reset, which will be described later. A3 indicates the output signal 33a of the subtraction circuit 14 having a magnitude obtained by subtracting the reference value 15 from the output signal 32a of the peak value hold circuit 13. Further, in A3, the original electrical signal 12a is shown by a broken line. Output signal 33 shown in A3
A and the original electrical signal 12a are compared by a comparator 16, and a pulse signal 34a of constant amplitude shown at A4 is generated from the comparator 16 in portions E and f that satisfy the above-mentioned comparison conditions.
If we look at column B in the same way, we can see that ``the electrical signal 12b is
1, and the output signal 32b of the peak value hold circuit 13 becomes as shown in B3.

Since the output signal 33b of the subtraction circuit 14 is smaller than the original electrical signal 12b as shown in B3, it does not satisfy the above-mentioned comparison condition, so there is no output from the comparator 16 as shown in B4. The output signal 34a of A4 is differentiated by the differentiator 17 to become a pulse signal 35a shown at C, and its negative signal resets the peak value hold circuit 13 at two points C and d.

The times C and d are the falling points of the pulse signal 34a of A4. This resetting in the middle of the scanning line 29a allows the study of the fluctuation range of the electrical signal 12a to be performed in the direction of the scanning direction, in order to examine the electric signal in the reaction zone that is in front of the agglomerate (left side in AO) and as close to the agglomerate as possible. This is necessary because the determination is performed based on the maximum value of the signal 12a. On the other hand, in column B, since there is no output signal from the comparator 16, it is reset by the constant scan completion signal 19 after the scan is completed. Comparator 16
Among the pulse signals 34a outputted from the integrator 20, those that proceed to the integrator 20 are processed as shown in A5 to A7.

That is, the pulse signal 34a shown at A4 is integrated by the pulse generation time by the integrator 20, and the pulse signal 34a shown at A4 is integrated into the integrated time signal 36 shown at A5.
a, and enters the comparator 21. A6 shows the integrated time signal 36a obtained over the entire scanning time T and the reference value 22.
When the cumulative time signal 36a exceeds the reference value 22, the comparator 21
A signal 23 indicating that there is aggregation is emitted. This A, ~A7
The determination method shown in is the method described in claim 1.

This determination method is based on the fact that when the tortoise signal 12a decreases from the previous maximum value to more than the reference value 15, it means that the light beam 4 is scanning the aggregate. It is based on the principle that the total time during which such a state occurs is proportional to the total area of the agglomerate, and it can be said to be an effective method for detecting the agglomerate state. In addition, in the above method, if the reference value 15 is set to a different value at the beginning and end of detection of each aggregate,
As shown in FIG. 4, it is possible to prevent an erroneous determination by the identification device based on minute fluctuations in the electrical signal corresponding to an agglomerate.

Fig. 4 D2 to D4 show cases of malfunction, and Fig. 4 E, to E
3 shows a signal waveform when normal operation is performed by changing the reference value 15.

For example, figure D.

In a reaction zone 2d where two aggregates are adjacent to each other with a minute interval 30 as shown in FIG.
In the electric signal 12d obtained at point d, a small peak value 31 corresponding to the interval 30 is formed at point g, as shown at point D.
When this electric signal 12d is processed by the apparatus shown in FIG. 2 in the same manner as in the above method, the following results are obtained.

That is, the output signal 32d of the heater value hold circuit 13 is D.
2, and is set at point g (described later). The output signal 33d of the subtraction circuit 14 is as shown in D3, and the pulse signal 34d of the comparator 16 is only a signal corresponding to the left agglomerate as shown in D4. This pulse signal 34d has the disadvantage that it does not correspond to the size of the aggregate. The heater value hold circuit 13 is overset by the differential signal at the falling edge (point g) of the pulse signal 34d. So now, when the reference value 15 is generated, when the pulse number 34d of D4 is generated, a certain value F (Fig. 4 E2) is obtained from the potential difference (Y-X).
If a small value is obtained by subtracting the value by E2, the subtraction circuit 14 outputs an output signal 33e shown as E2.

Comparing this output signal 33e with the original electrical signal 12d, we see that the pulse signal 3 corresponds to the size of the two aggregates without being affected by the presence of the small peak value 31, as shown in E3.
4e is emitted. The falling part of this pulse signal 34e (
The peak value hold circuit 13 is reset for the first time by the differential signal at point h). As with the reference value 15, the above-mentioned certain value F needs to be determined after careful consideration.

Note that the pulse signal 34e of E3 then proceeds to the integrator 20 and undergoes respective determinations.

The determination method to which the processing methods shown in D, E, to E3 are added is the method described in claim 2.

Note that, in order to actually operate the identification device shown in FIG. ) can be added.

As can be seen from the above examples, the method of the present invention optically and electrically determines the presence or absence of agglutination of a sample subjected to an agglutination reaction, so the determination time is short and the determination can always be made under constant conditions. This method has great effects, such as providing highly reliable judgments and eliminating individual differences unlike conventional methods, which are performed with the naked eye.

[Brief explanation of drawings]

1 and 2 schematically show an apparatus for carrying out the present invention, and the first
The figure is a schematic diagram of the scanning and photoelectric conversion device, FIG. 2 is a block diagram of the identification device, FIGS. 3A-C are explanatory diagrams showing specimen examples and scanning signals for each specimen, and FIGS. 4A-B is an explanatory diagram similar to FIG. 3. 2: reaction zone, 6: photoconductive element, 12, 12a, 12b,
12d: Electrical signal, 15: Reference value, 16, 21: Comparator, 22: Reference value.

Claims (1)

[Claims] 1. A specimen is scanned with a light beam having a minute cross-sectional area, and the light passing through the specimen is subjected to photoelectric conversion to extract an electrical signal of a size corresponding to the amount of transmitted light, and the electrical signal adjacent to the electrical signal is The method is characterized in that the time during which the range of variation between the local maximum value and the local minimum value exceeds the width of the reference value is integrated over the entire scanning time, and if this integrated time exceeds the reference time, it is determined that there is aggregation. A method for detecting the coagulation state of blood, etc. 2. Scan the specimen with a light beam with a minute cross-sectional area, perform photoelectric conversion on the light that has passed through the specimen, extract an electrical signal of a size corresponding to the amount of transmitted light, and calculate the adjacent maximum and minimum values of the electrical signal. In a method for detecting the state of aggregation of blood, etc., the time during which the range of fluctuations between , an aggregation of blood, etc., characterized in that an electrical signal is identified using different reference values when the electrical signal changes from a local maximum value to a local minimum value, and when it fluctuates finely in the local minimum value portion. How to detect the condition.