KR100973829B1 - Blood Typing Apparatus and Blood Typing Method Thereof - Google Patents

Abstract

The present invention relates to a blood diagnostic apparatus and a blood diagnostic method using the same. The blood diagnostic apparatus of the present invention comprises a passive element valve, a mixer part, and a chamber part. The passive element valve includes a first inlet through which the reaction reagent flows so that the reaction reagent and the blood flow into separate passages, and a second inlet through which blood flows. The mixer portion is connected to the first inlet and the second inlet, respectively, and the reaction reagent and the blood flow into the inner passage, and the reaction reagent and the blood flow along the inner structure installed in the inner passage and are mixed with each other. While the chamber unit is connected to the mixer unit, a space of a predetermined size is provided to contain the reaction reagent and blood passed through the mixer unit. Due to such a configuration, the blood diagnosis apparatus and the blood diagnosis method using the same of the present invention improve the coagulation reaction between the blood and the reaction reagents, and thus the blood type is more smoothly compared to the conventional manual blood discrimination test even with a very small amount of blood in the μl unit. Can be determined.

Blood, reaction reagents, determination, testing, mixing, purge, flocculation

Description

Blood diagnostic device and blood diagnostic method using the same {Blood Typing Apparatus and Blood Typing Method Thereof}

The present invention relates to a blood diagnostic apparatus and a blood diagnostic method using the same, and more particularly, to an improved blood diagnostic apparatus and a blood diagnostic method using the same to perform a blood test even with a very small amount of blood.

Blood is classified into several blood group groups, such as ABO and Rh blood groups. Human blood is distinguished from each blood type group to prevent side effects of blood transfusion caused by hemolysis of red blood cells when blood transfusion is needed, such as surgery. Blood type discrimination tests mix blood and reaction reagents, resulting in an antigen-antibody reaction of blood and reaction reagents.

Conventional blood group determination tests manually mix blood and reaction reagents on glass slides. Then, the aggregation reaction between the blood and the reaction reagent is observed under a microscope, and the blood type is visually determined. However, when a specific blood type or an irregular antibody is present, the aggregation reaction between the blood and the reaction reagent is not smoothly performed, and thus, it is difficult to determine the blood type by a conventional manual blood type determination test. As a result, a conventional blood test may be repeatedly performed in an emergency situation for surgery, or the reading time may be increased.

In addition, the conventional blood type determination test may use automated blood test equipment rather than manual work. Blood test automation equipment consists of erythrocyte sedimentation, centrifugal centrifugal process. Such blood test automation equipment has high accuracy of test results, but the equipment is large and relatively expensive compared to performance. Moreover, blood test automation equipment has the disadvantage of being difficult to carry or carry. Therefore, to date, even if the conventional blood type discrimination test is provided with blood test automation equipment even in a university hospital, the blood type discrimination test by hand is still widely performed.

The present invention has been proposed to solve the conventional problems as described above, and to provide a blood diagnostic apparatus that improves the aggregation reaction of blood and reaction reagents by actively mixing blood and reaction reagents compared to manual work. have.

Another object of the present invention is to provide a blood diagnosis method capable of more accurately determining a blood type even in the case of a very small amount of blood or in particular a blood type using a fuzzy algorithm program that models not only a blood diagnostic apparatus but also an expert's judgment process. There is this.

Blood diagnostic apparatus according to an embodiment of the present invention includes a passive element valve, a mixer (mixer), the chamber portion. The passive element valve includes a first inlet through which the reaction reagent flows, and a second inlet through which the blood flows, such that the reaction reagent and the blood flow into separate passages, respectively. The mixer unit is connected to the first inlet and the second inlet, respectively, and the reaction reagent and the blood flow into the inner passage, and the reaction reagent and the blood flow along the inner structure installed in the inner passage. Are mixed. The chamber part is connected to the mixer part, and a space having a predetermined size is provided to contain the reaction reagent and the blood passing through the mixer part.

The inner structure of the mixer unit includes a first spiral member twisted one or more revolutions in a clockwise direction, and a second spiral member twisted one or more revolutions in a counterclockwise direction while being connected to the first spiral member in succession. The first spiral member and the second spiral member are repeatedly formed while changing their order.

The inner structure of the mixer portion further includes a barrier member for changing the flow flow. The barrier member is formed on at least one of a surface of the first spiral member, a surface of the second spiral member, and an inner surface of the inner passage.

A reaction reagent passage is formed inside the first inlet, and a blood passage is formed inside the second inlet, and the reaction reagent passage and the blood passage are formed to be in contact with each end thereof to communicate with the internal passage. .

The passive element valve has a cross sectional area of the reaction reagent passage and a cross sectional area of the blood passage at the point where the reaction reagent passage and the blood passage are merged with one passage.

The chamber portion is formed such that the cross-sectional area of the inner space is gradually widened along the flow direction from the point connected to the inner passage of the mixer portion.

The cross section of the inner space is formed to be stepped sequentially from the point where the chamber portion is connected to the inner passage of the mixer portion.

In the blood diagnosis method according to an embodiment of the present invention, a blood diagnostic apparatus is used to mix blood with the reaction reagent and induce an aggregation reaction for a predetermined time. In addition, the blood diagnosis method analyzes an image in which the reaction reagent and the blood are coagulated, and applies the cohesive strength of the coagulated reaction image to a fuzzy rule to determine a blood antigen-antibody reaction according to the coagulation intensity. .

The blood diagnostic apparatus is manufactured by a micro-optical molding apparatus for curing light curable resin by irradiating light.

The image analysis step identifies the maximum size of the aggregated particles, the sharpness of the suspension, the total size of the aggregated particles, and the number of the aggregated particles in the aggregated reaction image observed by the microscope.

The clarity of the suspension converts the aggregated reacted image obtained through the microscope into a gray scale image file and quantifies the gray scale mean of the background excluding the aggregated particles in the gray scale image.

The blood discrimination step calculates cohesive intensity by applying input information about the coagulated reaction image to the fuzzy rule. The input information sets the maximum size (M) of the aggregated particles, the sharpness (C) of the suspension liquid, and the value (R) obtained by dividing the total size of the aggregated particles by the number of the aggregated particles based on three types.

The input information is divided into three sets of low, middle, and high according to the dilution concentration of blood, and weights the three sets. The input information is calculated by weighting three kinds of criteria according to the degree of determining the cohesive strength.

The blood discrimination step determines whether the calculated value of the aggregation intensity falls within a predetermined level range, and determines the blood antigen-antibody response based on the level.

Blood diagnostic apparatus and blood diagnostic method using the same according to an embodiment of the present invention by improving the aggregation reaction of the blood and the reaction reagent, even with a small amount of blood in μl more smoothly compared to the blood discrimination test by the conventional manual manual Can be determined. Because of this, the embodiment of the present invention has the advantage of being able to reduce the pain and discomfort of the patient due to repeated collection of blood, and also can be used for diagnosing blood of infants who are threatened with life by blood collection of a general adult.

In addition, embodiments of the present invention can quickly and easily discriminate blood types in which specific blood types or irregular antibodies are present, as compared with conventional manual blood discrimination tests. Therefore, the embodiment of the present invention has an advantage of minimizing medical accidents due to blood type misjudgment.

In addition, the embodiment of the present invention is not limited to the blood typing test, there is an advantage that can be applied to various immune serological tests using antigen-antibody reactions, such as hepatitis test, AIDS, syphilis.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art may easily implement the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

1 is a schematic diagram showing the components of a blood diagnostic apparatus according to an embodiment of the present invention, Figure 2 is a photograph showing a prototype of the blood diagnostic apparatus shown in FIG.

As shown in Figure 1 and 2, the blood diagnostic apparatus 100 according to an embodiment of the present invention is configured as follows to mix the blood and the reaction reagent more actively. As a result, the blood diagnosis apparatus 100 may further improve the aggregation reaction between the blood and the reaction reagent, and thus, the specific blood type or the irregular antibody may be more smoothly discriminated than in the related art. Such a blood diagnostic apparatus 100 is manufactured by, for example, micro-optical molding equipment as shown in FIG.

Figure 3 is a schematic diagram showing a micro-optical molding equipment for manufacturing the blood diagnostic apparatus shown in FIG.

As shown in FIG. 3, the micro-optic shaping equipment irradiates the light curable resin 60 with the light UV generated from the light source 10, so that the light curable resin 60 is irradiated with the light. By hardening, it manufactures with the blood diagnostic apparatus 100 of a predetermined shape.

Micro-optic shaping equipment includes a light source 10 as well as several optical system components. The optical component includes a filter 20 selectively passing according to the wavelength of light, and a shutter 30 positioned behind the filter 20 to adjust the irradiation amount of light by the opening and closing operations. In addition, the optical system component may further include a mirror 40 that changes the path of light propagation, and a focus lens 50 that focuses the light toward the photocurable resin 60.

In the micro-optical molding equipment, the elevator 80 behaves according to preset shape data according to the operation of the position control unit 70 in a state where the light irradiation point is fixed. The photocurable resin 60 positioned on the upper part of the elevator 80 may be manufactured in the shape of the blood diagnosis apparatus 100 while sequentially curing the light irradiated portion in a solid state. Here, the position controller 70 may be divided into the z stage 71 and the x-y stage 72. The z stage 71 moves the elevator 80 in the z axis direction, and the x-y stage 72 repositions the z stage 71 on the xy plane. The elevator 80 is positioned to be immersed below the liquid surface of the photocurable resin 60 by a predetermined thickness, and the first cross-sectional shape of the blood diagnosis apparatus 100 is primarily cured. After the molding of one layer is completed, the elevator 80 is lowered, thereby repeatedly curing the photocurable resin 60 in accordance with the cross-sectional shape of the blood diagnostic apparatus 100.

The blood diagnostic apparatus 100 manufactured as described above is divided into a passive element valve 110, a mixer 120, and a chamber 130. Hereinafter, the components of the blood diagnosis apparatus 100 will be described in more detail with reference to FIG. 1.

Passive element valve 110 is configured to have a separate passage for the reaction reagent and the blood, respectively, to control when the reaction reagent and blood is mixed. That is, the passive element valve 110 is divided into a first inlet 111 into which the reaction reagent is introduced, and a second inlet 112 into which blood is introduced. However, the first inlet 111 and the second inlet 112 may be different in size so as to control the amount of inlet to the reaction reagent or blood, but is formed in approximately the same shape.

The first inlet 111 is formed with a reaction reagent inlet 113 into which the reaction reagent is introduced from the outside, and a reaction reagent passage 114 extending from the reaction reagent inlet 113 to allow the reaction reagent to flow therein. The second inlet 112 is also formed by a blood inlet 115 through which blood is introduced from the outside, and a blood passage 116 extending from the blood inlet 115 to allow a reaction reagent to flow therein. The first inlet 111 and the second inlet 112 are formed to be inclined at a preset crossing angle θ ₁ so that the end portions of the respective passages come into contact with each other. That is, the first inlet 111 and the second inlet 112 induces the reaction reagent and blood to be separately input to be mixed with each other. The connecting portion 117 is connected to the first inlet 111 and the second inlet 112, respectively, and the reaction reagent passage 114 and the blood passage 116 are merged to have one internal passage. In this case, the cross-sectional area of the inner passage in the connecting portion 117 is preferably the same as the cross-sectional area of the reaction reagent passage 114 and the blood passage 116 is combined at the point where the reaction reagent and blood are mixed.

Due to this configuration, the passive element valve 110 mixes the reaction reagent 101 and the blood 102 while controlling the fluid flow to the reaction reagent 101 and the blood 102, respectively, as shown in FIG. You can. That is, the reaction reagent 101 is first introduced through the first inlet 111, but is stopped without exiting into the inner passage of the connecting portion 117 (FIG. 4A). That is, the reaction reagent 101 remains stationary due to the difference in internal pressure at the point where the area of the passage is sharply increased. This phenomenon can be easily understood through the capillary principle. As a next step, blood 102 flows through the second inlet 112 (FIG. 4B). Then, the passive element valve 110 flows the reaction reagent 101 and the blood 102 into the inner passage of the connecting portion 117 as the internal pressure increases.

The internal pressure in the passive element valve 110 is changed to a state as shown in FIG. 5 in consideration of the ratio of the width w and the height h of the passage. That is, the internal pressure in the passive element valve 110 maintains a negative pressure state lower than 0 in the a state of FIG. 4, and then changes to a positive pressure state in the b state of FIG. 4. ) And blood 102 will flow.

FIG. 6 is a perspective view illustrating an inside of a mixer of the blood diagnosis apparatus of FIG. 1.

As shown in FIG. 1 and FIG. 6, the mixer 120 is a member for mixing the reaction reagent 101 and the blood 102 introduced from the passive element valve 110 with each other. The mixer 120 has an internal passage 121 through which the reaction reagent 101 and the blood 102 can flow while communicating with the connection portion 117 of the passive element valve 110. In particular, the mixer 120 has a structure as shown in FIG. 6 in the inner passage 121, thereby improving the mixing effect of the reaction reagent 101 and the blood 102.

The internal structure of the mixer unit 120 includes a first spiral member 122 twisted at least one rotation in a clockwise direction and a second spiral member twisted at least one rotation in a counterclockwise direction while being connected to the first spiral member 122 in succession. It consists of 123. The first spiral member 122 and the second spiral member 123 are repeatedly formed in the longitudinal direction while changing their order. In addition, the internal structure of the mixer unit 120 further includes a barrier member 124 that further changes the flow flow. The barrier member 124 may be formed on each surface of the first spiral member 122 and the second spiral member 123 or may be formed on an inner surface of the inner passage. In the internal structure of the mixer unit 120, the more the first spiral member 122 and the second spiral member 123 are formed or longer in the longitudinal direction, the more the reaction reagent 101 and the blood 102 are mixed. Improve. The internal structure of the mixer unit 120 is a complicated three-dimensional form, but can be easily manufactured with the micro-optical molding equipment described above.

FIG. 7 is a photograph of a degree of mixing of blood and a reaction reagent before and after passing through a mixer of the blood diagnosis apparatus of FIG. 1.

As shown in FIGS. 4 and 7, the reaction reagent 101 and the blood 102 are introduced in a bisected state before passing through the mixer unit 120 (a in FIG. 7), but the mixer unit 120 is removed. It is also confirmed visually that the mixing degree became very high after passing (FIG. 7B). The reaction reagent 101 and the blood 102 are introduced into the chamber 130 in the state of the mixed material 103 while the agglomeration reaction occurs after passing through the mixer unit 120.

The chamber 130 is a space in which the mixed material 103 coming out of the mixer 120 is contained, and a space of a predetermined size or more is provided so that the mixed material 103 can be observed. The chamber unit 130 will be further described with reference to FIGS. 1 and 8.

FIG. 8 is a photograph illustrating an inflow form of the mixed substance in the chamber of the blood diagnosis apparatus of FIG. 1, and FIG. 9 is a schematic view illustrating an inflow form of the mixed substance in the chamber of the blood diagnosis apparatus of FIG. 8. .

As shown in FIGS. 1, 8, and 9, the chamber portions 130, 131, and 140 may have different flow forms depending on the shape of the internal space in which the mixed material 103 is contained. As a comparative example, if the chamber part 140 has a rectangular inner space shape as shown in FIG. 8A or FIG. 9A, the end of the fluid is maintained in an arch form by capillary pressure. That is, the chamber part 140 may suddenly increase the cross-sectional area of the passage, whereby the flow of the fluid may be stopped while the internal pressure is converted into a negative pressure value. For this reason, the bubble may generate | occur | produce in the chamber part 140, and it becomes difficult to observe aggregation reaction.

On the other hand, the chamber 130 according to the embodiment of the present invention is formed such that the cross-sectional area of the inner space is gradually increased as shown in (b) of FIG. 8 or (b) of FIG. That is, the chamber 130 is formed to be stepped sequentially in the flow direction of the fluid from the inner passage 121 of the mixer 120, thereby minimizing the change in the internal pressure. In addition, the chamber 130 according to an embodiment of the present invention is formed by inclining the inlet angle (θ ₂ ) of the fluid to a predetermined value or less in a form that is gradually wider as shown in FIG. To prevent the rapid diffusion of the fluid exiting 120.

In the following, in order to evaluate the performance of the blood diagnostic apparatus 100, the blood test results by the conventional manual labor and the blood test results using the blood diagnostic apparatus 100 was examined.

The conventional manual blood experiments were carried out according to the following steps.

① Dilution method using physiological saline prepares blood at 3% and reaction reagent (anti-A) at dilution rate of 25% and 10%, respectively.

② Drop the reaction reagent (3µl) onto the slide glass using a micro pipette.

③ Drop the blood (3 μl) into the reaction reagent using a micro pipette.

④ Mix blood and reaction reagent with a micro pipette.

⑤ Check the coagulation reaction of blood and reaction reagents within 2 minutes using a microscope.

On the other hand, the blood test using the blood diagnostic apparatus 100 according to an embodiment of the present invention was performed according to the following steps.

① Dilution method using physiological saline prepares blood at 3% and reaction reagent (anti-A) at dilution rate of 25% and 10%, respectively.

② Inject reaction reagent (3 μl) into the first inlet of the blood diagnosis apparatus 100 using a micro pipette. The reaction reagent then remains filled in the first inlet.

③ Inject blood (3µl) into the second inlet using a micro pipette. The internal pressure of the blood diagnostic device changes as the blood is injected, and the blood flows with the reaction reagent by capillary principle.

④ Switch blood diagnosis device to stand upright. The blood and the reaction reagent pass through the mixer section by capillary force and gravity and enter the chamber section.

⑤ Check the coagulation reaction of blood and reaction reagents within 2 minutes using a microscope.

FIG. 10 is a photograph of the results of ABO blood group experiment using a conventional blood discrimination test, and FIG. 11 is a photograph of the results of ABO blood group test using the blood diagnosis apparatus shown in FIG. 1.

In the blood discrimination picture shown in FIGS. 10 and 11, the aggregation of blood and the reaction reagent was examined for blood of A type. 3% diluted blood (3 μl) and 25% diluted reaction reagent (3 μl) were nearly similar to the aggregation in FIG. 10 (a) performed manually and in FIG. 11 (a) performed by the blood diagnostic apparatus. A response was observed to elicit. However, 3% diluted blood (3µl) and 10% diluted reaction reagent (3µl) were respectively shown in FIG. 10 (b) performed by hand and FIG. 11 (b) performed by the blood diagnostic apparatus, respectively. Although the aggregation reaction was induced, larger aggregated particles were observed in FIG. 11B using the blood diagnostic apparatus. That is, the blood diagnostic apparatus mixes blood and reaction reagents more effectively, thereby making it easier to discriminate blood by the aggregation reaction even with a small amount of reaction reagents.

Hereinafter, a blood diagnosis method using the blood diagnosis apparatus as described above will be described in more detail with reference to FIG. 12.

In the blood diagnosis method according to an embodiment of the present invention, as a first step, a reaction reagent and blood are mixed using a blood diagnosis apparatus, thereby inducing an aggregation reaction between the reaction reagent and blood (S1).

Then analyze the image (aggregated image) of the reaction reagent and blood (S2). In general, a medical professional can check the blood type based on the aggregated reaction image. However, the blood discrimination test may be difficult to identify by using a very small amount of blood, or aggregated reaction images in the presence of specific blood types or irregular antibodies.

In the blood diagnosis method according to an embodiment of the present invention, the cohesive strength is determined based on the coagulation reaction image as well as the coagulation of blood, and the blood type is determined through the coagulation intensity. This analysis of aggregated images is an image analysis program using matlab to grasp information such as maximum size of aggregated particles, clarity of suspended solids, total size of aggregated particles, and number of aggregated particles.

FIG. 13 is photographs converted into image files in analyzing the aggregated image illustrated in FIG. 12.

As shown in FIG. 13, the RGB image file obtained through a microscope (FIG. 13 a) is a gray scale image file (FIG. 13 b) and a binary image file (FIG. 13) for image analysis. Is converted to form c) of 13. In the gray scale image, the sharpness of the suspension may be quantified by obtaining the gray scale average of the background excluding the aggregated particles. In the binary image, the size and number of aggregated particles are calculated.

Then, the cohesive intensity is derived using the fuzzy rule, and the blood type is determined according to the cohesive intensity (S3). In other words, fuzzy rules are built on expert knowledge and experience, and cohesive strength is applied to these fuzzy rules. As shown in FIG. 14, the input information for determining the cohesive strength is subjected to a series of calculation processes through a fuzzy rule in a linguistic form, and is derived as output information indicating the level of cohesive strength.

Input information for determining the cohesive strength includes the maximum size of the aggregated particles, the clarity of the suspended liquid except the aggregated particles, and the total size of the aggregated particles. Divided by. This input information is derived from the image analysis program as above.

The sharpness of the suspension liquid, which is one of the input information, is determined as follows. If the aggregation reaction of blood and the reaction reagent occurs strongly, the suspension becomes transparent as the number of unaggregated red blood cells in the suspension decreases. Therefore, the sharpness of the suspension liquid grasps the average gray scale value of the background portion excluding the aggregated particles in the gray scale image file, and the stronger the aggregation reaction, the higher the gray scale value.

The maximum size of the aggregated particles is the most important input information in determining the cohesive strength of the blood. That is, the aggregated particles A1, B1, B2, and B3 shown in FIG. 15A, and the aggregated particles C1, C2, C3, and C4 shown in FIG. 15B are the same number. The sum of the total areas has the same condition. However, in the coagulation reaction in FIG. 15A, the coagulation reaction in FIG. 15B is the largest because the largest size of the coagulation particles A1 is one of the coagulation particles A1, B1, B2 and B3. It can be judged that it is stronger than the aggregation reaction.

The total area of the aggregated particles and the number of aggregated particles are not used as independent variables, but the ratio of the two variables is treated as one input information. The reason for treating the ratio of two variables as one input information will be described with reference to FIG. 16.

That is, since the size of the aggregated particles (M1 ~ M8) shown in (a) of Figure 16 is larger than the size of the aggregated particles (S1 ~ S4) shown in Figure 16 (c), (a) of FIG. It can be judged that the coagulation reaction in is stronger than the coagulation reaction in FIG. 16C. However, since the total area of the aggregated particles S1 to S4 shown in FIG. 16C is smaller than the total area of the aggregated particles M1 to M8 shown in FIG. 16A, the conclusion of the aggregation reaction is as follows. It is not desirable to derive In addition, the number of agglomerated particles L1 to L4 shown in FIG. 16B is the same as the number of agglomerated particles S1 to S4 shown in FIG. 16C, but the agglomerated particles M1 to M8. It can be concluded that a stronger aggregation reaction was induced in terms of overall size.

Therefore, the total area of the aggregated particles and the number of aggregated particles are treated as one input information and defined as a value obtained by dividing the total area of the aggregated particles by the number of aggregated particles. Such input information (total size of aggregated particles / number of aggregated particles) can be determined to be stronger as the numerical value becomes larger. Then, it can be judged that cohesive strength is roughly the strongest of FIG. 16B, and the weakest of FIG. 16C.

As such, it may be determined that the aggregation reaction is stronger as the maximum size of the aggregated particles is larger, the suspension is more transparent, and the larger the value obtained by dividing the total area of the aggregated particles by the number of aggregated particles. However, these input information is applied to the fuzzy rule, and is derived as output information of the level of the cohesive strength through a series of calculation processes.

The fuzzy rule is to determine whether the input information is included in the numerical set of linguistic concepts by regularizing the accumulated experience and knowledge of the experts into the numerical set of linguistic concepts. That is, the input information is divided into several sets according to the numerical range, and the degree of attribution to the set is represented in the form of a function. This type of function is defined as a membership function. At this time, the value of the function has a real value between 0 and 1. Each input information is divided into three sets of low, middle, and high, and is expressed as a function indicating the degree of attribution to it.

The membership function of the fuzzy rule was determined through actual blood experiments. The actual blood experiments used blood with three different dilution concentrations to determine the effectiveness of the program, and confirmed that the difference in the coagulation reaction according to the dilution concentrations showed different coagulation intensity levels. Here, the blood experiments used two different Type A blood samples, and each blood sample was diluted again to 3%, 2%, 1%. Blood experiments were repeated five times for each dilution concentration, with reaction reagents diluted to 50%. Therefore, 10 experimental data were collected for each dilution concentration.

The blood test image shown in (a) of FIG. 17 is a blood sample diluted to 1%, the blood test image shown to (b) of FIG. 17 is a blood sample diluted to 2%, and (c) of FIG. The blood experimental image shown in is a blood sample diluted to 3%. The coagulation reaction is clearly shown in the blood experiment image that the higher the dilution concentration of blood, the stronger the reaction.

The blood test image shown in FIG. 18 is only converted into a binary image file, and it is confirmed that the coagulation reaction is stronger as the dilution concentration of blood is higher. However, in the blood test image shown in FIG. 18, the size and number of aggregated particles are calculated according to an image analysis program.

The data of the input information calculated in this way may be graphed as shown in FIGS. 19 to 21, respectively. As can be seen more clearly in FIGS. 19 to 21, it is confirmed that the aggregation reaction is stronger as the dilution concentration of blood is higher.

The input information illustrated in FIGS. 19 to 21 may be re-expressed as a membership function indicating a degree of attribution to the set, and the membership function of the input information therefor is illustrated in FIGS. 21 to 24.

FIG. 22 is a graph showing the data on the maximum size of the aggregated particles shown in FIG. 19 as a membership function of the fuzzy rule, and FIG. 23 is a graph showing the data on the clarity of the suspension shown in FIG. 20 as a membership function of the fuzzy rule. 24 is a graph showing data on a value obtained by dividing the total area of the aggregated particles shown in FIG. 19 by the number of aggregated particles as a membership function of the fuzzy rule.

The data of the 3% diluted blood sample is used to construct a function corresponding to the "high" of the fuzzy set, and the 2% diluted blood sample constitutes the function corresponding to the "middle" of the purge set. Blood samples diluted 1% were used to construct a function corresponding to the "low" of the purge set.

Fuzzy reasoning is the process of inferring a new relationship or fact from a given fact or relationship. In other words, if it is defined as "IF x is A, THEN y is B", it is the process of dealing with the input "x is A" and the corresponding "how can y be obtained". Where A represents a fuzzy set of input information (maximum size of aggregated particles, sharpness of suspended solids, total area of aggregated particles divided by the number of aggregated particles), and B is a fuzzy set of cohesive strengths.

Since the input information of the three criteria differs in the degree of determining the cohesive strength, weights are set differently for each criterion. In other words, different weights are also set for sets divided into "low", "middle", and "high", so that the weights of the input information of the three criteria and the weights of the sets are respectively set. Addition and multiplication are used to determine the cohesive strength Table 1 shows the weights for the input information and the weights for each fuzzy set The weights of the input information determine how much the expert considers when visually determining the cohesive strength. The numerical value can also be changed according to the criterion for determining the cohesive strength, since the maximum size of the coagulated particles is the most important factor in determining the cohesive strength, so that the weight of the input information is set highest.

Input information Agglomerated particles
Maximum size (m) Suspension Clarity (C) Total Area of Aggregated Particles / Number of Aggregated Particles (R) 3 height height height 2 middle middle middle One lowness lowness lowness weight 0.8 0.3 0.6

M used for the purge rule is the maximum size of the aggregated particles, C is the sharpness of the suspension, R is the total area of the aggregated particles divided by the number of aggregated particles, L means the final cohesive strength. Cohesive strength is divided into Level 1, Level 2, and Level 3. If M, C, and R are all high conditions, the cohesive strength has a maximum value of 5.1, as calculated by Equation (1). If M, C, and R are all low conditions, the cohesive strength has a minimum value of 1.7, as calculated in Equation (2).

(0.8 × 3) + (0.03 × 3) + (0.6 × 3) = 5.1

(0.8 × 1) + (0.03 × 1) + (0.6 × 1) = 1.7

Then, under all conditions, the cohesive strength falls between 5.1 and 1.7. The cohesive strength calculated in this way is at level 1 if present between (1.7 and 2.8333), to level 2 if present between (2.8333 and 3.9667) and between (3.9667 and 5.1). Defined as level 3. The fuzzy rule receives the name of the input information and the type of the fuzzy set by the program generating the number of cases, and exists in the number of 27 cases as follows.

As a result, in the blood diagnosis method according to an embodiment of the present invention, blood types can be determined by defining blood types according to respective levels after finally calculating the aggregation intensity.

Examples of the present invention further evaluated the cohesive strength as follows. The higher the dilution concentration of blood, the greater the number of erythrocytes, the better the aggregation reaction. In other words, the higher the diluted concentration of blood, the larger the size of the aggregated particles. Therefore, the experimental results using 1% dilution, 2% dilution, and 3% dilution blood should correspond to level 1, level 2, and level 3, respectively, and apply them to the fuzzy rule to apply cohesive strength. Was calculated. However, in order to determine the actual weak A or weak type B blood, the number of red blood cells is fixed and the aggregation reaction is different depending on the amount of protein on the surface of red blood cells. However, because weak A or weak B is a specific blood type, it can be simulated by varying the blood dilution concentration. As shown in Table 2, the evaluation of the cohesive strength can be found to correspond appropriately to each level except for some cases due to blood experimental error. Here, the level value is a result of the calculation of the cohesive strength by the weight.

No 1% diluted blood 2% diluted blood 3% diluted blood level
(level) Level value
(level value) level
(level) Level value
(level value) level
(level) Level value
(level value) One One 2.19 2 3.4 3 4.52 2 One 2.18 2 3.37 3 4.58 3 One 2.18 2 3.99 2 3.38 4 One 2.18 2 3.19 2 3.47 5 One 2.18 2 4.14 3 4.57 6 One 2.22 2 3.41 3 4.51 7 One 2.14 2 3.07 2 3.56 8 One 2.21 2 3.37 2 3.89 9 One 2.16 2 3.39 2 3.48 10 One 2.12 2 3.37 3 3.98

In comparison with the second experiment and the tenth experiment in 3% diluted blood, the level was 3 and the same conclusion was obtained, but the level value was 4.58 and 3.98, respectively. Based on this, since level values have more differences within the same level, levels may be further subdivided as necessary.

That is, the preferred embodiments of the present invention have been described, but the present invention is not limited thereto, and various modifications and changes can be made within the scope of the claims, the detailed description of the invention, and the accompanying drawings. Naturally, it belongs to the range of.

1 is a schematic diagram showing the components of a blood diagnostic apparatus according to an embodiment of the present invention.

2 is a photograph showing a prototype of the blood diagnostic apparatus shown in FIG.

Figure 3 is a schematic diagram showing a micro-optical molding equipment for manufacturing the blood diagnostic apparatus shown in FIG.

Figure 4 is a schematic diagram showing each step of mixing the blood and the reaction reagents using the blood diagnostic apparatus shown in FIG.

FIG. 5 is a graph illustrating a pressure distribution in the blood diagnosis apparatus illustrated in FIG. 4.

FIG. 6 is a perspective view illustrating an inside of a mixer of the blood diagnosis apparatus of FIG. 1.

FIG. 7 is a photograph of a degree of mixing of blood and a reaction reagent before and after passing through a mixer of the blood diagnosis apparatus of FIG. 1.

8 is a photograph of the inflow form of the mixed material in the chamber of the blood diagnostic apparatus shown in FIG.

FIG. 9 is a schematic diagram illustrating an inflow form of a mixed material in the chamber part of the blood diagnosis apparatus illustrated in FIG. 8.

Figure 10 is a photograph of the results of the ABO blood type test using a conventional blood discrimination test by manual.

FIG. 11 is a photograph of an ABO blood group test result using the blood diagnosis apparatus shown in FIG. 1.

12 is a flowchart illustrating a blood diagnosis method according to an embodiment of the present invention.

FIG. 13 is photographs converted into image files in analyzing the aggregated image illustrated in FIG. 12.

FIG. 14 is a schematic diagram illustrating a relationship of determining cohesive strength using a fuzzy rule in determining cohesive strength shown in FIG. 12.

FIG. 15 is a diagram exemplarily illustrating a difference in cohesive strength according to a maximum size of coagulated particles among input information for determining cohesive strength shown in FIG. 12.

FIG. 16 is a diagram exemplarily illustrating a difference in the cohesive strength according to (the total area of the coagulated particles / the number of coagulated particles) among the input information for determining the cohesive strength shown in FIG. 12.

FIG. 17 is a blood test image of each blood sample tested for dilution concentrations to determine a membership function of the fuzzy rule shown in FIG. 12.

FIG. 18 is a blood test image obtained by converting the blood test image shown in FIG. 17 into a binary image file.

19 is a graph showing the maximum size of the aggregated particles calculated by the blood test images shown in FIG. 18 for each blood test result.

20 is a graph showing the sharpness of the suspended solids calculated by the blood test images shown in FIG. 18 for each blood test result.

FIG. 21 is a graph illustrating a value obtained by dividing the total area of the aggregated particles calculated by the blood test image shown in FIG. 18 by the number of aggregated particles for each blood test result.

FIG. 22 is a graph showing the data on the maximum size of the aggregated particles shown in FIG. 19 as a membership function of the fuzzy rule.

FIG. 23 is a graph showing the data on the sharpness of the suspension shown in FIG. 20 as a membership function of the fuzzy rule.

FIG. 24 is a graph showing data on a value obtained by dividing the total area of the aggregated particles shown in FIG. 19 by the number of aggregated particles as a membership function of the fuzzy rule.

<Description of the symbols for the main parts of the drawings>

100: blood diagnostic device 101: reaction reagents

102: blood 103: mixed substance

110: passive element valve 120: mixer

130 chamber portion (chamber)

Claims (14)

A passive element valve including a first inlet forming a reaction reagent inlet and a reaction reagent passage, and a second inlet disposed obliquely at a predetermined crossing angle with respect to the first inlet and forming a blood inlet and a blood passage; A mixer including an internal structure connected to an end of the reaction reagent passage and an end of the blood passage to form an internal passage through which the reaction reagent and the blood are supplied, the internal structure being installed in the internal passage to mix the reaction reagent and the blood with each other; )part; And A chamber part connected to the mixer part to provide a space having a predetermined size for storing a mixture of the reaction reagent and blood passed through the mixer part, And a cross-sectional area of the end portion of the inner passage in contact with the reaction reagent passage and the blood passage is equal to the sum of the cross-sectional area of the reaction reagent passage and the cross-sectional area of the blood passage. The method of claim 1, The inner structure of the mixer unit includes a first helix member twisted at least one rotation in the clockwise direction, and a second helix member twisted at least one rotation in a counterclockwise direction while being connected to the first helix member in succession. And the first spiral member and the second spiral member are repeatedly formed while changing their order. The method of claim 2, The inner structure of the mixer portion further comprises a barrier member for varying the flow flow, The barrier member is formed on any one or more of the surface of the first spiral member, the surface of the second spiral member, the inner surface of the inner passage. delete delete The method of claim 1, And the chamber portion is formed such that the cross-sectional area of the inner space is gradually widened along the flow direction from the point connected to the inner passage of the mixer portion. The method of claim 6, The chamber is a blood diagnostic apparatus that the cross-sectional shape of the inner space is formed step by step from the point connected to the inner passage of the mixer. Agglomeration that mixes the reaction reagent and blood and induces an agglutination reaction for a predetermined time using the blood diagnostic apparatus according to any one of claims 1, 2, 3, 6, and 7. Reaction step; An image analyzing step of analyzing an image in which the reaction reagent and the blood are aggregated and reacted; And And a blood discrimination step of determining a blood antigen-antibody reaction according to the aggregation intensity by applying the aggregation intensity of the aggregated reaction image to a fuzzy rule. The method of claim 8, The blood diagnostic apparatus is a blood diagnostic method using a blood diagnostic apparatus manufactured by a micro-optical molding equipment for curing light curable resin by irradiating light. The method of claim 8, The image analysis step is a blood diagnostic method using a blood diagnostic apparatus for grasping the maximum size of the aggregated particles, the clarity of the suspension, the total size of the aggregated particles, the number of the aggregated particles in the aggregated reaction images observed by the microscope. The method of claim 10, The clarity of the suspension is a blood diagnosis using a blood diagnostic apparatus that converts the aggregated reaction image obtained through a microscope into a gray scale image file and quantifies the gray scale average of the background excluding the aggregated particles in the gray scale image. Way. The method of claim 10, The blood discriminating step calculates the cohesive strength by applying the input information about the coagulated reaction image to the fuzzy rule, The input information includes blood for setting the maximum size (M) of the aggregated particles, the sharpness (C) of the suspension liquid, and the value (R) obtained by dividing the total size of the aggregated particles by the number of the aggregated particles based on three types. Method of diagnosing blood using a diagnostic device. 13. The method of claim 12, The input information is divided into three sets of low, middle, and high according to the dilution concentration of blood, and weights the three sets. And the input information is weighted to three kinds of criteria according to the degree of determining the cohesive strength to calculate the cohesive strength. The method of claim 13, The blood determination step is a blood diagnostic method using a blood diagnostic apparatus that determines whether the calculated value of the aggregation intensity is included in a predetermined level range, and determines the blood antigen-antibody response type by the level.

Images