WO2018190661A1 - Lateral flow assay sensor for comprising bridge structure for increasing detection sensitivity - Google Patents

Abstract

The present invention relates to a lateral flow assay sensor that increases detection sensitivity of the sensor by adding a bridge structure. The lateral flow assay sensor of the present invention is a lateral flow assay sensor that forms channels on a paper by using printing technology, comprising: a sample injection channel into which a sample is injected; a diagnosis channel spaced apart from one end of the sample injection channel by a predetermined distance so as to be elongated in the vertical direction, and having a diagnosis region coated with a detection material in the middle thereof; a bridge pad superimposed on each of the sample injection channel and the diagnosis channel so as to form a bridge for moving the sample from the sample injection channel to the diagnosis channel; and an absorption pad positioned on the top of the diagnosis channel to absorb a developing solution.

Description

Lateral flow analysis sensor with bridge structure for increased detection sensitivity

The present invention relates to a lateral flow analysis sensor by adding a bridge structure to increase the detection sensitivity of the sensor.

With the recent development of medicine and medical engineering, various causes of diseases and treatment methods are introduced, as well as more accurate diagnosis of various diseases is possible. Such accurate disease diagnosis enables patients to apply more effective and safe treatment methods, which is increasing the rate of disease treatment.

However, the diagnosis and treatment of the disease is mainly made in a specialized institution such as a hospital, there is a problem that the diagnosis target to be diagnosed, it is impossible to diagnose and diagnose the disease itself without visiting the hospital. However, it is difficult for a diagnosis subject or a patient to bear expensive test and treatment costs.

 In an effort to solve the above problems and difficulties, a telemedicin system has been introduced. This telemedicine system can be diagnosed by the patient or patient at a relatively low cost without visiting the hospital, but only the external abnormalities are diagnosed, and the diagnostic methods are also very limited. It is difficult to make an accurate diagnosis.

In particular, most of the diseases that can be a life-threatening problem are caused by infection with antigens such as pathogens. In the case of these diseases, prompt diagnosis is required. It is urgent to develop a device for diagnosis.

A biosensor is a device that can selectively measure a target material quickly by combining a signal conversion technology with a material that selectively reacts or binds to a specific target material. These biosensors have a variety of applications such as point-of-care, home diagnostics, agriculture / food / environmental measurement, biological terrorist measurement, industrial process monitoring, and research. Commonly used biosensors are molecules that diagnose viruses and various biomolecules using lateral flow immunoassay (LFA), polymerase chain reaction (PCR) based on lateral flow analysis using membranes. There is a biosensor.

The lateral flow immunoassay (LFA) sensor is widely used for on-site examinations such as pregnancy diagnosis or blood glucose measurement, and is widely used as a rapid medical diagnosis technology because of its advantages of easy and rapid analysis. Has been. In addition, the simplified flow sensor based on side flow immunoassay is being developed as a method for rapidly checking food safety and livestock health because the analysis cost is low and does not require expensive measuring instruments or specialized personnel.

Many related studies have been conducted on lateral flow immunoassay sensors, and commercial products have been released, but the sensitivity desensitization is a problem in many applications. Degradation is mainly due to the antigen-antibody response and the low efficiency of the reader quantifying it. In order to overcome these drawbacks, the development of biomarkers for efficient antigen-antibody reactions and the improvement of material functions of various pads and membranes are being conducted.

Accordingly, the present inventors add a bridge structure to the side flow analysis sensor in the course of studying a technique for increasing the detection sensitivity of the side flow immunoassay sensor, and through the added bridge structure, the analysis sample and the reaction solution are evenly mixed. , The present invention was completed by confirming that the precipitate generated during the reaction is separated and the detection sensitivity of the sensor is increased.

 In the prior art, Korean Patent No. 1262407 describes a lateral flow analysis measuring instrument including a bridge member, but the bridge of the prior art functions as a passage through which a test sample can flow or a blood separation filter. However, the effect of increasing the detection sensitivity by increasing the mixing ratio through the bridge structure of the present invention is not described at all.

It is an object of the present invention to provide an improved side flow analysis sensor which increases the detection sensitivity of the sensor by adding a bridge structure.

The side flow analysis sensor according to the present invention,

A side flow analysis sensor for forming a channel using a printing technique on paper, comprising: a sample input channel, which is a region into which a sample is input; A diagnostic channel spaced apart from one end of the sample input channel by a predetermined length and formed in a vertical direction, and having a diagnostic region coated thereon with a detection material therein; A bridge pad placed on the sample input channel and the diagnostic channel so as to overlap each other to form a bridge to move the sample of the sample input channel to the diagnostic channel; And an absorbent pad disposed on an upper end of the diagnostic channel to absorb the development solution.

In addition, the sample input channel may be formed as one channel and connected to a bridge pad, and the sample input channel may be configured as a sample pad into which a sample is input, and a conjugation pad containing a coloring material.

The sample input channel is further formed with a connecting portion connected to the bridge pad on the top, the width of the connecting portion is formed in the range of 0.3 ~ 0.7 times the width of the lower body portion of the sample input channel from the sample input channel to the bridge pad The sample can be mixed by diffusion while moving.

The sample input channel, the main body portion is formed in the width of 3 ~ 5mm and the length of 5 ~ 10mm, the connecting portion is formed to be narrowed to the width of 1 ~ 3mm in the length of 1 ~ 2mm at the upper end of the main body portion and the narrowed width 1 ~ Formed by 4 mm in length; The bridge pad has a width of 3 to 5 mm and a length of 3 to 7 mm; The diagnostic channel may have a width of 3 to 5 mm and a length of 10 to 15 mm, and the sample input channel and the diagnostic channel may be spaced at intervals of 2 to 4 mm.

In addition, the side flow analysis sensor of the present invention may be provided in a structure in which the sample input channel is formed into two channels and is joined to one channel on the upper side and then connected to the bridge pad.

In this case, the sample input channel is further formed with a connecting portion connected to the bridge pad on the upper portion of the two channels joined, the width of the connection portion is formed in the range of 0.3 ~ 0.7 times the width of one channel of the sample input channel By moving from the sample input channel to the bridge pad can be mixed by the sample diffusion.

The sample input channel, the first channel and the second channel is formed in a width of 3 ~ 5mm and a length of 3 ~ 7mm and formed side by side at intervals of 3 ~ 5mm, the first channel and the second channel has a top 2 ~ Joining in the length range of 5mm to form a narrow to 1 to 3mm, further extend the narrowed width to 1 to 4mm long to form a connection; The bridge pad has a width of 3 to 5 mm and a length of 3 to 7 mm; The diagnostic channel may have a width of 3 to 5 mm and a length of 10 to 15 mm, and the sample input channel and the diagnostic channel may be spaced at intervals of 2 to 4 mm.

The side flow analysis sensor according to the present invention,

When the mixed solution of the sample, the coloring material, the buffer, and the developing solution introduced into the sample input channel flows to the diagnostic channel, the flow is made through the bridge pad. In particular, the connection part of the sample input channel in contact with the bridge pad has a narrow width. Flowing from the narrow width to the wide width of the bridge pad allows diffusion to take place, further facilitating mixing. That is, turbulence is formed by laminar flow and sudden spreading by the same width of flow, thereby further promoting the mixing of the mixed solution.

In addition, the precipitate generated during the reaction by promoting the mixing is also generated enough to pass through the bridge pad section is filtered for the sediment in the bridge pad, and finally the mixed solution to be transferred to the diagnostic channel containing the foreign matter including the whole material The reliability of diagnosis can be improved by minimizing the increase of detection sensitivity.

Figure 1a is a plan view showing a one-channel side flow analysis sensor sample input channel according to a preferred embodiment of the present invention.

Figure 1b is a plan view showing a side flow analysis sensor according to another embodiment of a sample input channel according to the present invention.

Figure 2 is a plan view showing a side flow analysis sensor formed with two channels of the sample input channel in accordance with a preferred embodiment of the present invention.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the information provided herein is to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

1A is a plan view illustrating a one-channel side flow analysis sensor of a sample input channel according to a preferred embodiment of the present invention.

As shown, the side flow analysis sensor 10 formed of one sample input channel according to the present invention includes a sample input channel 20 which is a region into which a sample is input; A diagnostic channel 30 formed to be spaced apart from one end of the sample input channel in a vertical direction and formed long in the vertical direction, and having a diagnostic region 31 coated thereon with a detection material therein; A bridge pad 40 which forms a bridge by overlapping each of the sample input channel and the diagnostic channel to move the sample of the sample input channel to the diagnostic channel; It is configured to include; the absorbent pad 50 is placed on the upper end of the diagnostic channel to absorb the development solution.

The sample input channel 20 is a sample pad into which an analytical sample including an antigen-containing sample is input, and includes a reaction solution having a coloring material in the input assay sample, or additionally constitutes a separate conjugation pad. The sample input and the coloring material, which is a gold particle to which the antibody is bound, can be separately supplied. At this time, when the conjugation pad is additionally configured, as shown in FIG. 1B, the middle of the sample pad 21 is spaced apart in the longitudinal direction, and the conjugation pad 22 containing the coloring material is placed on the upper portion thereof, so that the sample pad While moving it can be combined with the coloring material to make the bond. In addition, the coloring material may be coated on the sample pad 21 to allow mixing to be performed while the injected sample is moved. In the present invention, a sample containing various buffer solutions and / or developing solutions is analyzed as a sample, and a solution containing a buffer solution and / or a developing solution as a coloring material is referred to as a reaction solution. It is expressed as a substance.

In the diagnostic channel 30, a sample flows using a nitrocellulose membrane, and a diagnostic region 31 having a diagnostic line 311 coated with a detection material is formed in the middle of the diagnostic channel. As the nitrocellulose membrane, NCk, NC94 and NC120 of Merck Millipore were used.

The bridge pad 40 has a predetermined length overlapping each other in the sample input channel 20 and the diagnostic channel 30 so as to connect a solution containing a sample and a coloring material injected into the sample input channel to the diagnostic channel. It is a pad. The bridge pad 40 is used to filter the foreign matter including the precipitate by the reaction of the sample is injected using an asymmetric membrane.

As the bridge pad, GX, GR, and GF in Vivid, an asymmetric membrane manufactured by PALL, may be used, and asymmetric super micron polysulfone having a pore size of 0.45 μm, 0.8 μm, and 8 μm; MMM) can be used to control the flow rate of the reactants and to filter the precipitate. In addition, the bridge pad can be used in a stacked form by selecting two kinds of the product can improve the separation effect and the detection strength of the precipitate.

On the other hand, the sample input channel 20 may be configured to be further connected to the bridge pad by further forming a connecting portion 24 which is a narrow section at the top. In this case, the bridge pad 40 may be formed to have a width wider than the width of the connection portion, for example, the width of the sample input channel before the width is narrowed, so that the bridge pad 40 may be connected to the diagnostic channel.

In this configuration, the mixed solution of the sample, the coloring material, the buffer, and the developing solution introduced into the sample input channel is moved to the bridge pad 40 while proceeding in a narrow width at the connection part 24, and at this time, because the width is suddenly expanded, The mixed solution is diffused and the mixed solution is better mixed by diffusion. That is, turbulent flow is formed by the laminar flow and the sudden widening of the flow of the same width to further promote the mixing of the mixed solution, and the precipitate generated during the reaction by the promotion of the mixing also includes the bridge pad 40 section. Filtering is performed in the bridge pad 40 so as to be sufficiently generated while passing through the mixture, and the mixed solution finally moved to the diagnosis channel 30 increases the detection sensitivity by minimizing the inclusion of foreign substances including precipitates, thereby increasing the diagnostic reliability. Can be improved.

In addition, the connection portion 24 is preferably formed in the range of 0.3 ~ 0.7 times the width of the connection portion 24 with respect to the width of the body portion 23 of the sample input channel so that the mixing by the sample diffusion. In the case of forming 0.3 times or less, the flow rate of the sample decreases, so that the diagnostic time is long, and when formed in 0.7 times or more, the width of the connection part within the above range is due to the lack of mixing effect due to diffusion due to the rapid width expansion. It is preferable to form

As the sample input channel as a preferable structure of the one-channel side flow analysis sensor,

The sample input channel 20 has a main body 23 having a width of 3 to 5 mm and a length of 5 to 10 mm, and the connecting part 24 has a length of 1 to 3 mm at a length of 1 to 2. mm at the upper end of the main body. It is formed by narrowing the width and further formed by narrowing the width 1 to 4mm in length; The bridge pad 40 has a width of 3 to 5 mm and a length of 3 to 7 mm; The diagnostic channel 30 is formed of a width of 3 ~ 5mm, and a length of 10 ~ 15mm; The sample input channel 20 and the diagnostic channel 30 may be configured to be spaced apart at 2 ~ 4mm intervals.

In this case, the gradually narrower width of the connection from the body portion of the sample input channel 20 to the connection portion removes the bent portion at right angles to prevent partial stagnation during solution flow to prevent the speed from being lowered. It is for.

In addition, when the width of the channel or pad is widened, the mixing effect can be seen to some extent, but there is a disadvantage in that the input amount of the sample is increased for diagnosis, and when the width is narrowed, the connection part 24 moves from the connection pad 24 to the bridge pad 40. Since the diffusion effect is reduced, it is preferable to configure the above range.

2, the sample input channel of the side flow analysis sensor of the present invention may be provided in two channels.

That is, the lower side into which the analysis sample or other solution containing the sample is introduced, and formed into two channels, and the upper side of the two channels may be combined into one to be connected to the bridge pad to allow the flow of the solution.

When the sample input channel 20 is formed into two channels, a solution containing a sample (analysis sample including a sample) is added to one side, and a solution containing a coloring substance (reaction solution containing a coloring substance) to the other side. ) Can be introduced into the flow while flowing into the bridge pad 40 finally joined.

At this time, since the diffusion is performed while moving from the connecting portion 24 to the bridge pad 40, the mixing is performed by diffusion, thereby generating a reaction precipitate by sufficient mixing to be separated from the bridge pad.

In addition, the conjugate pad may be placed in one of the two channels so that the development solution may be mixed with the analysis sample solution containing the sample of the generated material without the need for preparing and adding the coloring material. The conjugate pad settling method may be arranged in a bridge form by spaced apart some sections in any one channel of the two channels, or may be configured by overlapping without a spaced interval. In addition, it can be configured by coating the generating material on any one channel without the conjugate pad.

The sample input channel 20 composed of the two channels may also be connected to the bridge pad 40 by forming the connection part 24 on the upper portion of the two channels joined. The width of the connection part 24 may be formed in a range of 0.3 to 0.7 times with respect to the width of one channel of the sample input channel to be mixed by the sample diffusion while moving from the sample input channel to the bridge pad.

More specifically, the sample input channel 20,

The first channel 25 and the second channel 26 are formed in a width of 3 ~ 5mm and a length of 3 ~ 7mm and formed side by side at intervals of 3 ~ 5mm, the first channel and the second channel 2 ~ Joining in the length range of 5mm to form a narrower to the width of 1 ~ 3mm, and further extends the narrowed width to a length of 1 ~ 4mm to form a connection portion 24. When the channels are joined to be inclined at an angle of about 30 to 60 ° toward the center, it is possible to minimize the flow rate decrease due to the channel bending during the flow of the solution.

At this time, the bridge pad 40 is formed in a width of 3 ~ 5mm, and a length of 3 ~ 7mm. The bridge pad is formed to a size of 0.3 ~ 0.7 times the width of the connection portion of the sample input channel to obtain a diffusion effect.

In addition, the diagnostic channel 30 is formed with a width of 3 ~ 5mm, and a length of 10 ~ 15mm. The width is the same width as the other channels or pads except for the width of the connection portion of the sample input channel, the length is not limited to the above range can be varied by increasing the input amount of the developing solution.

In addition, the sample input channel 20 and the diagnostic channel 30 is spaced apart by 2 ~ 4mm intervals. When the thickness is less than 2 mm, the distance passing through the bridge pad 40 is short, and thus, the mixing reaction time may not be sufficiently provided. Since this does not generate a sediment by sufficient mixing while passing through the bridge pad and does not separate it, it is preferable that the sediment is supplied to the diagnostic channel to reduce the detection sensitivity. In addition, although 4 mm or more may be possible, the input amount of the developing solution is increased and the detection sensitivity is not improved.

Example

Experimental Example 1) Mixing confirmation of the sample and the reaction solution using the bridge structure

The degree of mixing with the assay sample (blood, urine, etc.) for detection and the reaction solution for the detection signal is one of the important factors in the diagnosis. Thus, the degree of mixing of the sample and the reaction solution through the bridge structure in the side flow analysis sensor of the present invention was confirmed.

The channel of the sensor is composed of nitrocellulose (NC) membrane, two sample input channels and one bridge structure, and the control has no bridge structure and two sample input channel structure sensors. Was used.

 Nitrocellulose membranes were NC75, NC94, NC120 from Merck Millipore, and glass pads (Glass fiber, 8964, 6613) from PALL, or GX, GR, and GF in Vivid were used as bridge pads.

One of the two sample input channels of the side flow analysis sensor of the control group containing no bridge structure and the side flow analysis sensor containing the bridge structure contained blue food coloring and a 1% (v / v) surfactant (Fitzgerald's surfactant 10G). In the other one, 60 μl of yellow food coloring and 1% (v / v) surfactant (Fitzgerald's surfactant 10G) were added in the same amount to allow fluid to flow, and the mixing of the two pigments was observed.

Figure 3 shows the results of confirming the structure of the side flow analysis sensor of the two sample input channels and the mixing of the fluid according to its structure. In the case of the control group without the bridge structure in the sensor (A), blue and yellow pigments were not mixed at all, whereas in the case of the sensor (B) with the bridge structure of the present invention, after passing through the bridge pad, the diagnostic channel In the blue and yellow pigments are mixed to give a green color, it was confirmed that the same result in the glass fiber or Vivid GX, GR, GF used as a bridge pad. This mixing of fluids is possible because the bridge pad used for the bridge structure facilitates diffusion of reactants by converting the laminar flow in the sensor into turbulent flow.

Through this, it was found that by applying the bridge structure in the side flow analysis sensor of the present invention, it is possible to promote the mixing of the analysis sample and the reaction solution in one sensor.

Experimental Example 2) Confirmation of increase in detection intensity through precipitation separation of reactants

The biggest problem of biosensors used in various diagnostics is a decrease in detection sensitivity. One of the causes of the deterioration of the detection sensitivity is the generation of noise and the reaction in the sensor due to the precipitate generated during the reaction between the analytical sample (blood, urine, etc.) for detection and the reaction liquid for the detection signal. Is impeding the flow of. Thus, it was confirmed that the detection intensity of the sensor is increased by separating the precipitate generated during the reaction using the bridge pad of the present invention.

Influenza virus protein and a side flow sensor capable of detecting the same were used to confirm the increase in detection intensity through precipitation separation of the reactants.

The sensor channel used NC75, NC94 and NC120 in the Nitrocellulose membrane membrane manufactured by Merck Millipore, and the bridge pad was an asymmetric membrane with 0.45㎛ GX, GR, GF and pore size of Vivid in PALL's products. Phosphorous asymmetric Super Micron Polysulfone (MMM) was used, and the conjugation pad was used by injecting the conjugation solution into the Vivid membrane used as the bridge pad without using the conjugation pad separately.

On the diagnostic line of the sensor, the antibody against Influenza A nucleoprotein was fixed at a concentration of 1 µg / ml, and in the bridge pad, a conjugate solution (RT dry 3 × AuNP-FIuA with PVP 28K 2%, 4% S10G) was injected and dried at room temperature and fixed. Induce the first reaction by injecting a sample (0.1M Tris-HCl (pH 8.5), 1% Triton X-100) containing the influenza A nucleoprotein at a concentration of 100 ng / ml into the sample input channel. , A gold enhancement reagent (0.1M Tris-HCl (pH 8.5) base in H ₂ SO ₄ -10mM H ₃ NO) can be used to increase the detection signal of gold particles in the secondary reaction. To induce a secondary reaction.

The sample is introduced into the sample input channel of the sensor and flows to induce the first reaction to combine the influenza A nuclear protein with the influenza A nuclear protein of the diagnostic line, and then to the signal-increasing reagent and influenza A nuclear protein introduced for the second reaction. The bound gold particles (AuNP-FIuA) reacted to generate a color reaction on the diagnostic line, and the color intensity was measured.

As shown in FIG. 4, in the case of a sensor using a 0.45 μm MMM alone as a bridge pad, the detection emphasis is similar to that of a sensor composed only of a nitrocellulose membrane without a bridge structure (sensor 1, sensor 9, and sensor 17). On the other hand, the detection intensity was increased for the sensor (Sensors 3 to 8, Sensors 11 to 16, and Sensors 19 to 24) that used Vivid alone or Vivid and 0.45 μm MMM as a bridgepad. In particular, it was confirmed that NC94 was used as the nitrocellulose membrane, and the detection intensity of the sensor including the bridge pad structure was increased the most.

In addition, comparing the detection intensity of the sensor with Vivid alone and the sensor with Vivid and 0.45㎛ MMM as a bridge pad, it was confirmed that the detection intensity of the sensor with Vivid and 0.45㎛ MMM was higher than the sensor with Vivid only. .

Through this, it can be predicted that the asymmetric super micron polysulfone (MMM) increases the separation efficiency of the precipitate and, in the case of Vivid, increases the detection strength of the sensor by increasing the activity of the reaction solution for increasing the detection strength.

Experimental Example  3) Bridge structure  Containing Lateral flow analysis  On the sensor Bridge pad  Check flow rate control of fluid according to type and composition

The change of the flow velocity of the fluid according to the type and configuration of the bridge pad used in the side flow analysis sensor including the bridge structure was confirmed.

The sensor channel was NC180 from Merck Millipore's nitrocellulose (NC) membrane, and there was one sample input channel and a bridge structure. The control group did not have a bridge structure and one sample input. A sensor (control group 2) using a kind of sample pad was manufactured and used as a channel structure type sensor (control group 1) and a bridge pad. As a bridge pad of the sensor of the present invention, asymmetric super micron polysulfone (MMM) having a pore size of 0.1, 0.2, 0.45, 0.8, 8, 10, 20 μm was used as an asymmetric membrane. In this case, in the case of the asymmetric membrane, since the pore sizes of the upper and lower surfaces of the membrane are different, the pore size of the asymmetric membrane surface in contact with the channel may affect the flow of the fluid.

Blue food coloring and 1% (v) in the sample input channel of the control group 1, which is a sensor that does not include the bridge structure and the control pad 2, which is a sensor using the absorbent pad 22 (AP22) as a bridge pad, and the sensor including the bridge structure. / v) After the surfactant (Fitzgerald company surfactant 10G) was added to allow the fluid to flow, it was measured the time that all the channels of the sensor is dyed. In this case, the case where the surface of the asymmetric membrane with the small pore contacted the channel of the sensor was low, and the case where the surface of the asymmetric membrane with the large pore size contacted the sensor channel was indicated as High.

Figure 5 shows the results of confirming the flow rate of the fluid in the sensor according to the type and configuration of the bridge pad used in the bridge structure of the present invention. As a result of checking the flow rate of the fluid in the sensor during the same time period (A), when the low pore size of the asymmetric membrane is in contact with the sensor channel, the fluid is bridged in the sensor using the 0.1 μm MMM membrane as the bridge pad. Failure to pass through the pads was observed, and the flow of fluid was significantly slowed even with MMM membranes of 0.2, 0.45, and 0.8 μm. On the other hand, it was confirmed that the effect of the pore size of the MMM membrane does not appear much in the case of the high surface where the large pore size of the asymmetric membrane is in contact with the channel of the sensor.

In addition, as a result of measuring the time for staining all the channels of the sensor (B), it was confirmed that the flow rate of the fluid varies depending on the type of the asymmetric membrane and the pore size of the asymmetric membrane in contact with the channel of the sensor.

Through this, it can be seen that the flow of the fluid can be adjusted according to the type of the bridge pad in the side flow analysis sensor including the bridge structure of the present invention and the surface of the bridge pad in contact with the channel of the sensor.

Experimental Example 4) Confirmation of the increase in the detection sensitivity by increasing the mixing of the assay sample and the reaction solution

It was confirmed that the increase of the mixing of the analysis sample and the reaction solution by the bridge structure increases the detection sensitivity.

The sensor channel was NC120 and NC180 in the nitrocellulose (NC) membrane of Merck Millipore, and there was one sample input channel, and it was made in the form including the bridge structure. A sample input channel structure type sensor was manufactured and used. Vivid GF membrane was used as asymmetric membrane of PALL as a bridge pad.

Serum containing CRP (c-reactive protein) is injected into the sample input channel of the sensor and flows. Serum passes through a conjugate pad to which gold particles coated with antibodies to CRP are fixed, and the CRP and gold in serum The CRP antibody coated on the particles was allowed to bind. CRP and the CRP antibody coated on the gold particles are combined to flow the diagnostic channel of the sensor, and when combined with another CRP antibody fixed to the diagnostic line, the gold particles bound to the CRP on the diagnostic line Color development occurred and the color intensity was measured.

Figure 6 shows the result of comparing the detection intensity of the sensor with the bridge structure of the present invention and the sensor of the control group. Looking at the color development (A) and the detection intensity (B) of the diagnostic line when using a serum containing 100ng / ㎖ CRP, compared to the control sensor not containing a bridge structure of the sensor comprising the bridge structure of the present invention The detection intensity was increased, and it was confirmed that the detection intensity of the sensor including the bridge structure was increased compared to the sensor without the bridge structure, regardless of the type of nitrocellulose membrane used as the sensor channel.

In addition, when the detection intensity is compared to the type of nitrocellulose membrane, both the control sensor not including the bridge structure and the sensor including the bridge structure of the present invention are detected more than the case of using NC180 in the nitrocellulose membrane. It was confirmed that the strength was high.

In addition, the color of the channel according to the concentration of the CRP (0, 0.1, 1, 10, 100ng / ㎖) using a sensor consisting of NC120 and looking at the detection intensity (D) of the control group does not include a bridge structure It was confirmed that the detection intensity of the sensor including the bridge structure of the present invention is higher than that of the sensor.

Through this, it was found that the side flow analysis sensor including the bridge structure of the present invention can increase the detection intensity by promoting the mixing of the analysis sample and the reaction solution by the bridge structure.

Claims (8)

1. In the side flow analysis sensor to form a channel using a printing technology on paper,

   A sample input channel 20 which is a region into which a sample is input;

   A diagnostic channel 30 formed to be spaced apart from one end of the sample input channel in a vertical direction and formed long in the vertical direction, and having a diagnostic region 31 coated thereon with a detection material therein;

   A bridge pad 40 which forms a bridge by overlapping the sample input channel 20 and the diagnostic channel 30 so as to move the sample of the sample input channel to the diagnostic channel;

   And an absorbent pad (50) placed on the upper end of the diagnostic channel to absorb the development solution.
2. The method of claim 1,

   The sample input channel 20 is formed in one channel, side flow analysis sensor, characterized in that connected to the bridge pad.
3. The method of claim 2,

   The sample input channel (20) is a side flow analysis sensor, characterized in that composed of a sample pad (21) into which a sample is input, and a conjugation pad (22) containing a coloring material.
4. The method of claim 2,

   The sample input channel 20 further includes a connection part 24 connected to the bridge pad 40 at an upper end thereof, and the width of the connection part is 0.3 to 0.7 times the width of the lower main body part 23 of the sample input channel. Formed in the range of the side flow analysis sensor characterized in that the mixing by the sample diffusion is made while moving from the sample input channel to the bridge pad.
5. The method of claim 4, wherein

   The sample input channel 20, the body portion 23 is formed of a width of 3 ~ 5mm and a length of 5 ~ 10mm, the connecting portion 24 is a width of 1 ~ 3mm in the length of 1 ~ 2mm at the top of the main body portion It is formed to be narrowed to form a narrower width of 1 ~ 4mm further;

   The bridge pad 40 has a width of 3 to 5 mm and a length of 3 to 7 mm;

   The diagnostic channel 30 is formed of a width of 3 ~ 5mm, a length of 10 ~ 15mm,

   The sample input channel 20 and the diagnostic channel 30 is a side flow analysis sensor, characterized in that spaced at intervals of 2 ~ 4mm.
6. The method of claim 1,

   The sample input channel 20 is formed in two channels, the side flow analysis sensor, characterized in that joined to one channel on the upper side and then connected to the bridge pad.
7. The method of claim 6,

   The sample input channel 20 further includes a connection part 24 connected to the bridge pad at the upper end of the portion where the two channels are joined, and the width of the connection part is 0.3 to 0.7 with respect to one channel width of the sample input channel. Side flow analysis sensor, characterized in that formed by the range of the doubling from the sample input channel to the bridge pad to be mixed by the sample diffusion.
8. The method of claim 7, wherein

   The sample input channel 20,

   The first channel 25 and the second channel 26 are formed in a width of 3 ~ 5mm and a length of 3 ~ 7mm and formed side by side at intervals of 3 ~ 5mm,

   The first channel 25 and the second channel 26 are joined in the length range of 2 to 5 mm to form a narrow width of 1 to 3 mm, and further extends the narrowed width to a length of 1 to 4 mm. );

   The bridge pad 40 has a width of 3 to 5 mm and a length of 3 to 7 mm;

   The diagnostic channel 30 is formed of a width of 3 ~ 5mm, a length of 10 ~ 15mm,

   The sample input channel 20 and the diagnostic channel 30 is a side flow analysis sensor, characterized in that spaced at intervals of 2 ~ 4mm.

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