WO2021177548A1

WO2021177548A1 - Fully automatic lateral flow immunoassay kit and measuring device for measuring multiple biomarkers of saliva

Info

Publication number: WO2021177548A1
Application number: PCT/KR2020/016121
Authority: WO
Inventors: 유성덕
Original assignee: 주식회사 이바이오젠
Priority date: 2020-03-03
Filing date: 2020-11-17
Publication date: 2021-09-10
Also published as: KR102156569B1

Abstract

The present invention relates to a fully automatic lateral flow immunoassay strip automatic analysis apparatus for measuring multiple biomarkers from saliva, the apparatus comprising a measuring device and a lateral flow immunoassay (hereinafter, LFA) kit, that is an LFA kit. The lateral flow immunoassay strip automatic analysis apparatus according to the present invention has, in connection with LFA strip reaction determination, data stored regarding the coloring ratio between a test line and a control line with regard to each concentration by using a sample for each target substance concentration. When the coloring ratio between a test line and a control line is imaged through a camera as a result of a reaction between a sample with an unknown target substance concentration and a LFA strip, and calculated through image processing, same is compared with existing data such that the concentration of the target substance in the sample is calculated and displayed, thereby enabling fast, highly reproducible, and highly sensitive analysis.

Description

Fully automatic lateral flow immunoassay kit and measuring device for the measurement of multiple biomarkers in saliva

The present invention relates to an automatic side flow immunoassay strip analysis device, and more particularly, to a side flow immunoassay strip automatic analysis device for measuring multiple biomarkers in saliva and a method for detecting multiple biomarkers from saliva using the same it's about

Lateral Flow Immuno Assay (LFA) is an analysis technology that can detect a sample (target material) in an unknown sample through a sandwich immunoassay technique using nanoparticles and a sample flow using a membrane.

The diagnostic strip used in the lateral flow immunoassay includes an elongated rectangular support made of an adhesive plastic material, and a sample pad, a conjugate pad, a detection pad, and an absorption pad, which are disposed approximately sequentially from one side to the other on the support. is made including

The lateral flow immunoassay method is used for diagnosis in various medical/environmental fields or for non-medical self-testing because the analysis principle is simple, the analysis time is short, and the production cost is low.

In particular, the lateral flow immunoassay is widely used for field analysis because it is inexpensive, easy to carry and fast detection, and can be easily used by the general public without specialized skills.

The side-flow immunoassay is largely divided into a sandwich assay method using primary and secondary antibodies against a target substance and a competitive assay method using only a single antibody.

A representative example of using the sandwich assay method of lateral flow immunoassay is urine chorionic gonadotropin (hCG), such as "a kit for diagnosing a cow's pregnancy and a method for diagnosing a pregnancy using the kit" of Korean Patent Registration No. 10-0930517. It is a pregnancy diagnostic kit that collects from and detects whether you are pregnant.

In the sandwich assay lateral flow analysis method, hCG injected into the sample pad flows along the membrane (detection pad) by capillary action while binding to the gold nanoparticle-antibody conjugate immobilized on the conjugate pad.

At this time, when the gold nanoparticles, which are detection indicators (probes), develop color due to the binding of the secondary antibody immobilized to the detection region, the result can be confirmed.

In the sandwich assay method, the side-flow immunoassay method visually evaluates the detection factors by the color development of gold nanoparticles that form an immunocomplex with the target material.

The side-flow immunoassay of the competitive assay method is mainly used to detect small-molecular substances, and a representative example is a drug test kit.

In the competitive assay lateral flow analysis method, the target material injected into the sample pad flows along the membrane (detection pad) by capillary action while binding to the gold nanoparticle-antibody conjugate immobilized on the conjugate pad.

At this time, the gold nanoparticle-antibody conjugate to which the target material is bound cannot bind to the target material-protein complex immobilized in the detection region, and accordingly, the gold nanoparticles, which are detection indicators (probes), do not collect on the detection line, resulting in negative color development. weaker than in the case of the sample.

The side-flow immunoassay of the competitive assay method evaluates the detection factors by visually confirming that the gold nanoparticles, which form an immunocomplex with the target substance, cannot bind to the detection line, resulting in weak or no color development.

For precise diagnosis and detection of detection substances, a reading system capable of realizing high-sensitivity analysis and quantitative analysis through improved analysis technology is required.

However, the lateral flow immunoassay method is not excellent in analytical sensitivity as it relies on visual identification evaluation, and there is a problem in that quantitative analysis is difficult.

In addition, the conventional side-flow immunoassay has low measurable sensitivity, so it is difficult to measure a sample requiring higher sensitivity.

In addition, in the case of a sample requiring quantitative analysis, analysis technology cannot be applied.

The present invention was invented to improve the above problems. Instead of using only one side of the side flow immunoassay strip, more detection strips can be mounted, and a plurality of detection strips can be detected using the internal rotation device and detection device. An object of the present invention is to provide a fully automatic lateral flow immunoassay kit and measuring device that can detect substances more accurately and conveniently.

In addition, the present invention stores the data on the color development ratio of the control line and the test line for each concentration using the sample for each concentration of the target material in determining the LFA strip reaction, and the reaction between the sample and the LFA strip whose concentration of the target material is unknown. In this way, when the color development ratio of the control line and the test line is taken through the camera and calculated through image processing, the concentration of the target substance in the sample is calculated and displayed by comparing it with the existing data. is to provide

In addition, the present invention easily detects and analyzes a target material regardless of skill level by using the fully automatic side flow immunoassay kit and measuring device for measuring multiple biomarkers in saliva according to the present invention, and a series of detection and analysis processes This is to provide a measurement method that can significantly improve the efficiency of analysis by automatically performing

In order to achieve the above object, the present invention is a sample collection unit comprising a cotton for collecting a sample including saliva in the oral cavity of the subject to be measured; A polygonal columnar body having an internal space into which the cotton for collection is inserted and having a plurality of reaction surfaces in the vertical direction, and a lateral flow immunoassay (hereinafter referred to as Lateral Flow ImmunoAssay) provided in plurality at the lower side of the plurality of reaction surfaces, respectively a strip for LFA) and an LFA cartridge accommodating the lower part of the main body and including a reaction container in which the reaction solution to be reacted with the sample discharged from the cotton for collection is accommodated; And it is possible to provide a side flow immunoassay kit for measuring multiple biomarkers in saliva, characterized in that it comprises a reaction inducing unit for allowing the reaction solution from which the sample is eluted and the LFA strip to react.

In addition, the present invention is a measurement having an internal space capable of accommodating a lateral flow immunoassay (hereinafter referred to as LFA) cartridge comprising a polygonal column-shaped body and a reaction vessel accommodating the lower part of the body and containing a reaction solution. housing; a drive transmission unit for generating a driving force for forward and reverse rotation of the docking station; a CCD camera mounted on one side of the measuring housing to photograph a test line formed by the reaction solution reacting to the LFA strip provided on the lower side of the plurality of reaction surfaces provided in the vertical direction of the body; and a processing unit that calculates the concentration of the target substance in the sample by comparing and analyzing the reaction determination data stored in advance from the test line photographed with the CCD camera. It may be possible to provide a fully automatic measuring device for measuring.

In addition, the present invention, a first step of collecting a sample including saliva in the oral cavity of the measurement subject with a cotton for collection; A second step of eluting the sample into the reaction solution in the LFA cartridge by putting the cotton for collection into the lateral flow immunoassay (LFA) cartridge seated in the measurement housing; After the first time elapses, the cotton for collection is applied to the LFA cartridge so that the reaction solution from which the sample is eluted is reacted with a plurality of strips for LFA respectively formed on a plurality of reaction surfaces provided on the polygonal columnar body of the LFA cartridge. A third step of lowering further into the inside or opening the stopper formed in the reaction vessel containing the reaction solution to which the sample discharged from the collecting cotton reacts, and contacting the reaction solution with the plurality of strips for LFA; After a second time period greater than the first time has elapsed, the LFA cartridge is rotated and stopped so that the plurality of reaction surfaces face the CCD camera provided in the measurement housing, and the control line and test line of the LFA strip are connected to the CCD camera. A fourth step of taking a picture; And the image processor calculates the concentration of the target material in the sample by comparing and analyzing the image captured by the CCD camera with the reaction determination data stored in the color development ratio between the control line and the test line, and the calculated result value is measured It is also possible to provide a method for detecting a target substance using a fully automatic measuring device that measures multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that it includes a fifth step of outputting through a display panel formed on one side of the housing. is of course

According to the present invention having the configuration as described above, the following effects can be achieved.

First of all, the present invention does not utilize only a single side of the LFA strip, unlike the prior art and existing products, and provides a detection strip for each reaction surface of the polygonal prism-shaped body, so that more detection strips can be mounted. It has the advantage of being able to detect a substance.

In addition, the present invention is provided in the measurement housing through the drive transmission unit for rotating and elevating the LFA cartridge and the sample collection unit, and the processing unit for calculating the concentration of the target material compared to the prior art and the existing products more diverse and more target material It has the advantage of being able to detect more accurately, quickly and conveniently.

In addition, the measuring device according to the present invention stores data on the color development ratio of the control line and the test line for each concentration by using the sample for each concentration of the target material in determining the reaction of the LFA strip, and measures the concentration of the target material. By photographing the color development ratio of the control line and the test line with the camera due to the reaction between the unknown sample and the LFA strip, and comparing it with the existing data through the processing of the image processor, the concentration of the target material in the sample is calculated and displayed directly through the display panel. Therefore, it has the advantage that rapid, high reproducibility, and high-sensitivity analysis are possible.

1 is a conceptual diagram schematically illustrating the overall structure of an LFA kit according to an embodiment of the present invention and a fully automatic measuring device for measuring multiple biomarkers of saliva with this kit.

Figure 2 is a conceptual diagram schematically showing the structure of the LFA kit according to an embodiment of the present invention.

3 is a conceptual diagram schematically showing the structure of an LFA kit according to another embodiment of the present invention.

4 is a conceptual diagram schematically illustrating the overall structure of a fully automatic measuring device for measuring multiple biomarkers of saliva according to an embodiment of the present invention.

5 is a conceptual diagram schematically illustrating the overall structure of a fully automatic measuring device for measuring multiple biomarkers of saliva according to another embodiment of the present invention.

6 is a conceptual diagram schematically illustrating the overall structure of an LFA kit according to another embodiment of the present invention and a fully automatic measuring device for measuring multiple biomarkers of saliva with this kit.

7 is a conceptual diagram schematically showing the overall structure of the LFA kit according to another embodiment of the present invention.

8 is a conceptual diagram schematically illustrating the overall structure of an LFA kit according to another embodiment of the present invention and a fully automatic measuring device for measuring multiple biomarkers of saliva with this kit.

9 and 10 are conceptual views schematically showing the overall structure of the LFA kit according to another embodiment of the present invention.

11 is a block diagram sequentially illustrating a method for detecting a target material using a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit according to an embodiment of the present invention.

12 is a conceptual diagram sequentially illustrating a method for detecting a target material using a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit according to an embodiment of the present invention.

Advantages and features of the present invention, and methods for achieving them, will become apparent with reference to the embodiments described below in detail in conjunction with the accompanying drawings.

However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various different forms.

In the present specification, the present embodiment is provided to complete the disclosure of the present invention, and to fully inform those of ordinary skill in the art to which the present invention pertains to the scope of the present invention.

And the invention is only defined by the scope of the claims.

Accordingly, in some embodiments, well-known components, well-known operations, and well-known techniques have not been specifically described to avoid obscuring the present invention.

In addition, like reference numerals refer to like elements throughout the specification, and terms used (referred to) herein are for the purpose of describing the embodiments and are not intended to limit the present invention.

In this specification, the singular also includes the plural unless specifically stated otherwise in the phrase, and elements and operations referred to as 'comprising (or having)' do not exclude the presence or addition of one or more other elements and operations. .

Unless otherwise defined, all terms (including technical and scientific terms) used herein may be used with the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs.

Also, terms defined in commonly used dictionary are not to be interpreted ideally or excessively unless defined.

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings.

First, FIG. 1 is a conceptual diagram schematically illustrating the overall structure of an LFA kit according to an embodiment of the present invention and a fully automatic measuring device for measuring multiple biomarkers in saliva with this kit.

And, Figure 2 is a conceptual diagram schematically showing the structure of the LFA kit 100 according to an embodiment of the present invention.

And, Figure 3 is a conceptual diagram schematically showing the structure of the LFA kit 100 according to another embodiment of the present invention.

4 is a conceptual diagram schematically illustrating the overall structure of the fully automatic measuring device 200 for measuring multiple biomarkers of saliva according to an embodiment of the present invention.

And, FIG. 5 is a conceptual diagram schematically illustrating the overall structure of a fully automatic measuring device for measuring multiple biomarkers of saliva according to another embodiment of the present invention.

And, Figure 6 is a conceptual diagram schematically showing the overall structure of the LFA kit 100 according to another embodiment of the present invention and a fully automatic measuring device for measuring multiple biomarkers of saliva with this.

And, Figure 7 is a conceptual diagram schematically showing the overall structure of the LFA kit 100 according to another embodiment of the present invention.

And, FIG. 8 is a conceptual diagram schematically illustrating the overall structure of the LFA kit 100 according to another embodiment of the present invention and a fully automatic measuring device for measuring multiple biomarkers of saliva with this.

And, Figure 9 and Figure 10 is a conceptual diagram schematically showing the overall structure of the LFA kit 100 according to another embodiment of the present invention.

In addition, FIG. 12 is a conceptual diagram sequentially illustrating a method for detecting a target material using a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit according to an embodiment of the present invention.

For reference, in FIGS. 7 and 9 , reference numeral 117 denotes a buffer container for saliva.

It can be seen that the present invention has a structure including a Lateral Flow Immuno Assay (LFA) kit, that is, an LFA kit 100 and a measuring device 200, as shown.

When looking at the LFA kit 100 with reference to FIGS. 2 and 3 along with FIG. 1 , it can be seen that the LFA kit 100 has a structure including a sample collecting unit 110 , an LFA cartridge 120 , and a reaction inducing unit 130 .

First, the sample collection unit 110 includes a cotton 111 for collection from which a sample including saliva is collected in the oral cavity of a measurement subject.

The LFA cartridge 120 has an internal space into which the collecting cotton 111 is inserted, and a polygonal column-shaped body 121 having a plurality of reaction surfaces 121r in the vertical direction, and a plurality of reaction surfaces 121r lower It includes a strip 122 for LFA provided in plurality on the side, and a reaction vessel 123 that accommodates the lower portion of the body 121 and accommodates the reaction solution in which the sample discharged from the cotton 111 for collection is reacted. .

The reaction inducing unit 130 reacts with the reaction solution from which the sample is eluted and the strip 122 for LFA.

It goes without saying that the present invention can be applied to the above-described embodiments, and can also be applied to the following various embodiments.

First, the sample collecting unit 110 may further include a first rod 112a having a lower end to which the collecting cotton 111 is mounted and an upper end coupled to the fixing member 250 of the measuring device.

In addition, the sample collection unit 110 may further include a handle 113 provided at the upper end of the rod 112 and having an upper surface in which a coupling groove 113g coupled to the fixing member 250 of the measuring device is recessed. .

In addition, the sample collection unit 110 may further include a separation preventing piece 114 extending from the outer peripheral surface of the lower side of the first rod 112a and regulating the separation of the cotton 111 for collection to the upper side.

Also, the sample collecting unit 110 may further include a male thread 115 formed along the outer peripheral surface of the lower end of the first rod 112a and detachably coupled to the stopper 131 .

Here, it is also possible to grasp that the collecting cotton 111 is coupled through the lower end of the first rod 112a having the male thread 115 formed therein.

At this time, the upper surface of the collecting cotton 111 is in contact with the lower surface of the separation prevention piece 114, and the lower end of the first rod 112a is detachably coupled with the stopper 131 of the reaction inducing unit 130. 111) is preferably exposed from the lower surface.

The main body 121 may be provided in the shape of a polygonal prism manufactured by adding or subtracting the number of the response surfaces 121r according to the type of measurable target material such as a triangular prism, a quadrangular prism, or a hexagonal prism.

In addition, the reaction vessel 123 may have an inner surface facing the plurality of reaction surfaces 121r on which the LFA strip 122 is mounted.

That is, a buffer for saliva, which is a reaction solution in which the collected sample is dissolved, flows out from the lower end of the body 121 through the reaction inducing unit 130 and reacts with the LFA strip 122 in the reaction vessel 123 ) so that the inner surface faces the reaction surface 121r, that is, it is preferable that the lower side of the main body 121 is accommodated in the reaction vessel 123 .

On the other hand, the reaction inducing unit 130, as shown in Fig. 2 (b), is built in the lower side of the body 121 and a solution capsule 132 in which the reaction solution is accommodated, and the upper and lower caps (131u, 131d) further comprising Embodiments of the structure may be applied.

The upper cap 131u constitutes the stopper 131, has an upper surface in contact with the lower surface of the cotton 111 for collection, is coupled to the lower end of the first rod 112a, and seals the upper portion of the solution capsule 132 will do

The lower cap 131d constitutes the stopper 131, is disposed on the lower surface of the solution capsule 132, and passes through the upper cap 131u according to the descending and rising of the first rod 112a. It engages with the lower end of 112a) to open the lower surface of the solution capsule 132.

On the other hand, the LFA cartridge 120 may further include a guide channel 124 passing through the center of the main body 121 as shown in FIG. 3 and having an inner surface corresponding to the outer surface shape of the handle 113 .

Here, the center of the guide channel 124, the first rod 112a, and the lower cap 131d is arranged on a straight line as shown in the dashed-dotted line in FIG. 3, the lower end of the first rod 112a and the lower cap 131d ) is necessary for the precise positional combination of

That is, the upper cap 131u is basically fixed to the first rod 112a by the male thread 115, and is rotated and pulled out from the lower end of the first rod 112a when collecting a sample including saliva. It can be said that it is disposable and discarded.

In addition, the lower cap 131d is made of a material such as elastic rubber, synthetic rubber, and synthetic resin and is engaged with the male thread 115 and then as the first rod 112a rises from the lower surface of the solution capsule 132 . It is separated so that the reaction solution flows out into the reaction vessel (123).

In addition, the reaction inducing part 130 is formed in the center of the upper surface of the lower cap 131d, and rotates and descends while further comprising a fastening groove (not shown) having an inner circumferential surface with a female thread that engages with the male thread 115. may be

In other words, the lower end male thread 115 of the first rod 112a is in the form of a bolt, and the first rod 112a descends into the solution capsule 132 and moves to the upper surface of the lower cap 131d. After that, if the lower cap (131d) is engaged with the fastening groove of the lower cap (131d) to continue to rotate and descend.

Thereafter, when the lower end of the first rod 112a moves to a position where the first rod 112a is no longer screw-rotated, that is, to the bottom dead center, a measuring device to be described later automatically raises the first rod 112a. Accordingly, the lower cap 131d opens the lower surface of the solution capsule 132 , and the reaction solution from which the sample is eluted flows into the reaction vessel 123 .

At this time, since the cotton 111 for collection is contracted in the process of being fixed to the lower cap 131d according to the screw rotation, the sample including saliva is effectively eluted to the reaction solution side, and reaches the bottom dead center and automatically rises again. In the process, the measuring device to be described later continues to rise after repeatedly raising and lowering the first rod 112a several times.

By repeating the rising and falling of the first rod 112a in this way, the reaction solution is more uniformly mixed with the sample, and the reaction with the strip 122 for LFA can be induced to be reliably made.

On the other hand, the measuring device 200, again with reference to FIG. 4 together with FIG. 1, a polygonal column-shaped body 121 and a reaction vessel 123 that accommodates the lower portion of the main body 121 and accommodates the reaction solution. It may include a measurement housing 210 having an internal space that can accommodate the LFA cartridge 120 including the.

The measurement housing 210 may further include an indicator 211 capable of displaying the current analysis progress.

Indicator 211, for example, 'Mixing', 'Binding', 'Analyzing', 'Finishing', etc., step by step, so that the measurer can grasp the current progress, the corresponding part is turned on in a bar shape with an LED. Various applications and deformation designs are available. Of course it is possible.

Meanwhile, the measurement housing 210 may further include a wired charging terminal 212 for connection with an external power source or various power sources for stable power supply.

In addition, the measuring device 200 includes a drive transmission unit 220 that generates a driving force for rotating the joint head 260 and the docking station 270 forward or backward, or raising or lowering the joint head 260 . can do.

The joint head 260 has a lower surface provided with a fixing member 250 that is detachably coupled to the upper end of the first rod 112a.

The docking station 270 is built in the measurement housing 210 so that the LFA cartridge 120 is mounted so that it can rotate forward and reverse.

In addition, the measuring device 200 is mounted on one side of the measuring housing 210 and reacted to the LFA strip 122 provided on the lower side of the plurality of reaction surfaces 121r provided in the vertical direction of the main body 121 . A camera 230 for photographing the test line 301 formed by the reaction solution may be included.

Of course, the camera 230 may further include a light source 231 such as a flash to obtain a clean and clear image.

The camera 230 may be a CMOS or CCD camera, but is not limited thereto.

In addition, the measuring device 200 may include a processing unit 240 for calculating the concentration of the target material in the sample by comparing and analyzing the reaction determination data stored in the test line 301 photographed by the camera 230 .

On the other hand, the measuring device 200 is provided on one side of the measuring housing 210, is electrically connected to the drive transmission unit 220, the camera 230, and the processing unit 240, the pre-stored reaction determination data and the test line ( The controller 280 including the image processor 281 for calculating the color development ratio of 301 may be further provided.

In addition, the measuring device 200 is provided on one side of the measuring housing 210, is electrically connected to the controller 280, and further includes a start switch 282 for controlling the on/off of the power and the operation of the controller 280. may be provided.

In addition, the measurement device 200 is provided on one side of the measurement housing 210, is electrically connected to the start switch 282 and the controller 280, and outputs an analysis result through the image processor 281 (display panel ( 290) may be further provided.

Meanwhile, the drive transmission unit 220 is provided on the upper side of the measurement housing 210 and is connected to the joint head 260 to rotate the first motor 221 and the joint head 260 . ) and may include a second motor 222 for raising or lowering the joint head 260 by being connected.

In addition, the drive transmission unit 220 is provided on one side of the measurement housing 210 , is connected to the docking station 270 , and rotates the polygonal column-shaped body 121 seated in the docking station 270 by a predetermined angle. It may include a third motor 223 for generating a driving force to stop and stop.

Here, in the repeated operation of raising and lowering the first rod 112a by the second motor 222 , the lower cap 131d of the reaction inducing unit 130 as described above is connected to the lower end of the first rod 112a. Combined, the reaction solution in the solution capsule 132 is more uniformly mixed with the sample to reliably react with the LFA strip 122 .

The above-described predetermined angle is a pattern in which n response surfaces 121r of the main body 121 having a polygonal column shape are rotated and stopped by 360/n degrees to face the camera 230 provided in the measurement housing 210, n -1 or n repetitions.

Here, n is a positive integer of 3 or more.

For example, when the main body 121 has a quadrangular pole shape, the pattern of rotating and stopping by 360/4, that is, 90 degrees so that the four response surfaces 121r face the camera 230 can be repeated 3 or 4 times. will be able

Meanwhile, in the present invention, the measurement housing 210 may further include a power button 283 capable of turning on/off the power of the entire device together with the start switch 282 as shown in FIG. 5 unlike FIGS. 1 to 4 .

Here, of course, the measurement housing 210 may include a wired charging terminal 212 capable of supplying power through a USB cable together with the function of a wired charging terminal.

At this time, one side of the measurement housing 210 may further include a display panel 290 as shown in FIG. 5 as in the embodiment of FIGS. 1 to 4 , and the test mentioned in the embodiment of FIGS. 1 to 4 . Although the line 301 and the control line 302 to be described later are displayed only in the form of a line, in the present embodiment, the test line and the control line may be displayed in the above-described line form, as well as being displayed in various forms as follows. can

That is, in the embodiment shown in FIGS. 5 to 10, which will be described later, including FIG. 5, any shape and shape that can enhance visual recognition, such as a spot, pixel, dot shape, along with the aforementioned line shape, is applied. You can do it.

In addition, the color developing material of the lines, spots, pixels, and dots of the test line and the control line described above is gold (Au), quantum dots, colored glass or colored plastic beads, fluorescent materials, enzymes, enzymatic reactions or non-enzymatic reactions. Of course, it may be any one or a combination of at least one or more selected from the group consisting of a chromogenic substrate used for

On the other hand, the present invention may be applied to an embodiment of the structure using the cotton swab 116 instead of the first rod 112a as shown in FIGS. 5 to 10 .

That is, the sample collection unit 110 may further include a cotton swab 116 having a lower end to which the cotton collection 111 is mounted.

Here, the reaction inducing unit 130 is a second rod (112b) having a lower end accommodated in the reaction vessel 123 and an upper end coupled to the fixing member 250 of the measuring device when viewed with reference to FIGS. 5 to 7, It is mounted on the lower side of the second rod 112b and may further include a first impeller 133a for stirring the reaction solution so that the sample discharged from the cotton 111 for collection reacts with the reaction solution.

At this time, on the upper surface of the main body 121, as shown in FIG. 7(a), a rod hole 121a through which the second rod 112b can enter and exit, and a first input hole ( 121b) may be further provided.

In addition, the drive transmission unit 220 is shown with reference to FIGS. 5 to 7 , the first, second, and third motors 221 , together with the fixing member 250 (refer to the structure of FIGS. 222, 223) may be further provided.

Here, in the second rod 112b having a lower end accommodated in the reaction vessel 123 and an upper end coupled to the clamp 250 of the measuring device, the fixing member 250 is detachably coupled to the upper end of the second rod 112b. will become

The joint head 260 has a lower surface provided with a fixing member 250 .

The first motor 221 is provided on the upper side of the measurement housing 210 and is connected to the joint head 260 to rotate the joint head 260 .

The second motor 222 is connected to the joint head 260 to raise or lower the joint head 260 .

The third motor 223 is provided on one side of the measurement housing 210, is connected to the docking station 270, and rotates and stops the polygonal column-shaped body 121 seated in the docking station 270 by a predetermined angle. to generate driving force.

Here, the first motor 221 may grasp that the first impeller 133a mounted on the lower side of the second rod 112b rotates.

Here, n is a positive integer of 3 or more.

Meanwhile, the reaction inducing unit 130 may further include a stopper 131 capable of opening and closing the upper surface of the reaction vessel 123 , referring to FIGS. 8 to 10 .

In addition, the reaction inducing unit 130 may further include a second impeller 133b mounted on the upper side of the stopper 131 to stir the reaction solution so that the sample discharged from the cotton 111 for collection reacts with the reaction solution. There will be.

In addition, of course, the reaction inducing unit 130 may further include a third rod 134 having both upper and lower ends connected to the stopper 131 and the bottom surface of the reaction vessel 123 .

Accordingly, the stopper 131 is rotated upward or rotated together with the reaction vessel 123 as shown in FIG. 8(b) by the drive transmission unit 220 including the docking station 270 on which the reaction vessel 123 is seated. While descending, the upper surface of the reaction vessel 123 will be opened and closed.

In addition, a second input hole 121c may be further provided on the upper surface of the main body 121 so that a saliva buffer or a cotton swab 116 can be inserted as shown in FIG. 9( a ).

In addition, of course, the main body 121 may further include a collection guide hopper 118 formed in a funnel shape as shown in FIG. 10 .

Here, the collection guide hopper 118 is provided to prevent the solution from jumping out of the body 121 during high-speed rotation when extracting a sample from the cotton 111 for collection of the cotton swab 116 .

At this time, the collection guide hopper 118 is required, when the cotton 111 for collection is inserted into the body 121, and in the state in which the cotton 111 for collection is inserted into the body 121 (b) When the stopper 131 is opened and the cotton swab 116 containing the cotton 111 for collection is lifted as shown, the sample is removed from the cotton 111 for collection in contact with the lower end of the funnel, that is, the lower portion of the collection guide hopper 118. This is because it can achieve a squeezing effect.

This is because the sample extraction efficiency can be improved by squeezing the sample from the cotton 111 for collection, and when the sample extraction efficiency is increased, the detection reaction time can be shortened.

Meanwhile, the drive transmission unit 220 may further include a fourth motor 224 and a fifth motor 225 , referring to FIGS. 8 and 9 .

The fourth motor 224 is provided on one side of the measurement housing 210, is connected to the docking station 270, and rotates the circular or polygonal pole-shaped body 121 seated on the docking station 270 by a predetermined angle. It stops and generates a driving force capable of forward and reverse rotation.

At the same time, the fourth motor is connected to the lower end of the third rod 112c in the third rod 112c that extends to the stopper 131 that can open and close the upper surface of the reaction vessel 123 and is accommodated in the reaction vessel 123 . It will also be able to perform a role of forward and reverse rotation of the third rod (112c).

In addition, the fifth motor 225 serves to generate a driving force for raising or lowering the body 121 seated in the docking station 270 .

Accordingly, the fourth motor 224 may grasp that the second impeller 133b mounted on the upper side of the stopper 131 rotates.

The above-described predetermined angle is a pattern in which the n response surfaces 121r of the main body 121 having a circular or polygonal column shape rotate and stop by 360/n degrees to face the camera 230 provided in the measurement housing 210 . may be repeated n-1 times or n times.

Here, n is a positive integer of 3 or more.

For example, when the main body 121 has a circular or polygonal prism shape, the pattern of rotating and stopping by 360/4, that is, 90 degrees so that the four response surfaces 121r face the camera 230 is performed three or four times can be repeated.

Hereinafter, a method for detecting a target material using a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit according to various embodiments of the present invention will be described with reference to FIGS. 11 and 12 .

For reference, reference numerals not indicated in FIGS. 11 and 12 refer to FIGS. 1 to 10 .

First, a sample including saliva is collected in the oral cavity of the subject to be measured with the cotton 111 for collection mounted on the lower end of the first rod 112a (S1: first step).

Next, the first rod 112a is pushed into the LFA cartridge 120 seated in the measurement housing 210 as shown in FIG. , the sample is eluted in the reaction solution in the LFA cartridge 120 (S2: second step).

Then, so that the reaction solution from which the sample is eluted is reacted with the plurality of LFA strips 122 respectively formed on the plurality of reaction surfaces 121r provided in the polygonal columnar body 121 of the LFA cartridge 120, the first After time has elapsed, the rod 112 is further lowered into the LFA cartridge 120 as shown in FIG. The stopper 131 is opened to bring the reaction solution into contact with the plurality of LFA strips 122 (S3: third step).

Subsequently, the LFA cartridge 120 is rotated and stopped so that the plurality of reaction surfaces 121r face the camera 230 provided in the measurement housing 210 after a second time period greater than the first time has elapsed to stop the LFA strip ( 122), the control line 302 and the test line 301 are photographed by the camera 230 (S4: fourth step).

Then, the image processor 281 compares and analyzes the image captured by the camera 230 with the reaction determination data stored in the color ratio between the control line 302 and the test line 301 to calculate the concentration of the target substance in the sample. and output the calculation result through the display panel 290 formed on one side of the measurement housing 210 (S5: fifth step).

Here, although it is variable depending on the type of target material or the analysis situation, the first time is 0.1 seconds to 5 minutes, and the second time may be appropriately adjusted within 1 second to 15 minutes.

At this time, in the third step (S3), the second motor 222 raises or lowers the handle 113 connected to the fixing member 250 and the coupling groove 113g as shown in FIG. Of course, it is also possible to additionally carry out the operation of accelerating the reaction of the reaction solution.

The LFA (Lateral Flow Immuno Assay) strip automatic analysis apparatus according to the present invention includes an LFA kit including a saliva collection swab and a reaction solvent, a measurement and analysis device, and a wired charger.

In the LFA (Lateral Flow Immuno Assay) strip automatic analysis apparatus according to the present invention, data on the color development ratio of the control line and the test line for each concentration is stored using the sample for each concentration of the target material in determining the LFA strip reaction, and the target Due to the reaction of the LFA strip with the sample whose concentration is unknown, the color development ratio of the control line and the test line is photographed through a camera (eg, CMOS or CCD camera) and calculated through image processing. The concentration of the target substance in it is calculated and displayed.

In the LFA (Lateral Flow Immuno Assay) strip automatic analysis device according to the present invention, 1) After collecting saliva, a cotton swab is inserted into the LFA kit, mounted on the measuring device, and when the “Start” button is pressed, the reaction and analysis are performed automatically and the result is confirmed 2) saliva on the cotton swab is effectively eluted and mixed with the reaction solvent and can be used continuously as long as there is no device failure, the analysis kit is a consumable, 5) the measuring device includes a drive transmission unit to drive the LFA cartridge, 6) the camera (e.g., CMOS or CCD camera) and flash are installed, and 7) the back side of the measuring device has an analysis result confirmation window and an instrument panel to check the progress.

Hereinafter, one embodiment of the present invention will be described.

The embodiments described herein are exemplary preferred embodiments, and the technical spirit of the present invention is not limited thereto.

<Example 1> Preparation of LFA (Lateral Flow Immuno Assay) strip automatic analysis device

The LFA strip cartridge was configured to mount two LFA strips on each side as a square face. The LFA strip cartridge has a swab for collecting the sample inside, and when the swab for collecting the sample is inserted from the top down, the upper layer of the reaction solution container surrounded by the inner membrane is pierced and the sample is placed on the swab. The attached sample is dissolved in the reaction solution, and after a certain period of time, if you push the handle of the cotton swab from top to bottom once more, the membrane on the lower layer of the reaction solution container is pierced and the reaction solution in which the sample is dissolved flows out into the container at the bottom of the cartridge and the LFA strip installed designed to react with

The sample collection part had a handle and was composed of a cotton swab structure with a pointed tip. The sampling part was made in a pointed shape so that it could easily penetrate the upper and lower layers of the reaction solution container inside the cartridge.

The camera for analysis consisted of a light source and a CCD camera to photograph the color development of the control line and test line of the LFA strip.

The kit mounting part and rotation driving part consisted of an LFA strip cartridge mounting part and a rotating part that the cartridge can rotate. The kit mounting part and rotation driving part were made to automatically rotate one side after a certain period of time after the reaction of the sample and the LFA strip so that the camera can photograph the LFA strip located on each side of the quadrangular plane.

The control unit is composed of a board that controls the CCD camera, light source, rotation unit, LCD window, power supply and progress indicator. The control unit is designed to enable image processing to obtain the color ratio between the control line and the test line.

The power supply and observation part are composed of power button, LCD window and progress indicator.

<Example 2> Operation of LFA (Lateral Flow Immuno Assay) strip automatic analysis device

After turning on the device's power switch, a sample was collected using a sampling swab, the swab was pushed into the cartridge, and the start button was pressed.

The cotton swab moves into the solvent container and moves up and down several times, and as the cotton swab rotates, it is fixed with the lower stopper. There was a hole in the pit, and the mixed solution flowed out into the reaction vessel.

The solution is applied to the LFA kit strip and the reaction is performed, and the reaction result is displayed as a line on the membrane. When the reaction is completed, the camera of the device takes the membrane to obtain an image. After shooting 4 sides, the results of the lines from each side were analyzed and the reaction results were displayed in the result window. At this time, the image processor digitizes the color development ratio of the control line and the test line in the captured image and compares the color development ratio of the existing control line/test line to find the concentration of the target substance in the sample, and the target concentration appears on the LCD window ended

As described above, it can be seen that the present invention has a basic technical idea to provide a fully automatic lateral flow immunoassay kit and measuring device for the measurement of multiple biomarkers in saliva capable of detecting various target substances.

And, within the scope of the basic technical spirit of the present invention, many other modifications and applications are also possible for those of ordinary skill in the art.

Claims

a sample collection unit including a cotton for collection from which a sample including saliva is collected in the oral cavity of the subject;

A polygonal columnar body having an internal space into which the cotton for collection is inserted and having a plurality of reaction surfaces in the vertical direction, and a lateral flow immunoassay (hereinafter referred to as Lateral Flow ImmunoAssay) provided in plurality at the lower side of the plurality of reaction surfaces, respectively a strip for LFA) and an LFA cartridge accommodating the lower part of the main body and including a reaction container in which the reaction solution to be reacted with the sample discharged from the cotton for collection is accommodated; and

A side flow immunoassay kit for measuring multiple biomarkers in saliva, characterized in that it comprises a reaction inducing part for allowing the reaction solution from which the sample is eluted and the LFA strip to react.
The method according to claim 1,

The sample collection unit,

A first rod having a lower end to which the collecting cotton is mounted and an upper end coupled to a fixing member of the measuring device;

a handle provided at the upper end of the first rod and having an upper surface in which a coupling groove coupled to the fixing member of the measuring device is recessed;

A separation prevention piece extending from the outer peripheral surface of the lower side of the first rod and regulating the removal of the cotton for collecting to the upper side;

It is formed along the outer circumferential surface of the lower end of the first rod and further includes a male thread coupled to and detached from the stopper formed in the reaction vessel,

The side flow immunoassay kit for measuring multiple biomarkers in saliva, characterized in that the cotton for collection is penetrated through the lower end of the first rod having the male thread formed thereon.
The method according to claim 1,

The sample collection unit,

Side flow immunoassay kit for measuring multiple biomarkers of saliva, characterized in that it further comprises a cotton swab having a lower end to which the cotton for collection is mounted.
The method according to claim 1,

The reaction inducing unit,

In the first rod having a lower end to which the collecting cotton is mounted and an upper end coupled to a fixing member of the measuring device, a stopper formed in the lower end of the first rod and the reaction vessel;

a solution capsule built in the lower side of the body and containing the reaction solution;

An upper cap constituting the stopper, which has an upper surface in contact with the lower surface of the cotton for collection, is coupled to the lower end of the first rod, and seals the upper portion of the solution capsule;

A lower cap constituting the stopper, which is disposed on the lower surface of the solution capsule and engages with the lower end of the first rod penetrating the upper cap according to the descending and rising of the first rod to open the lower surface of the solution capsule. further comprising,

The side flow immunoassay kit for measuring multiple biomarkers in saliva, characterized in that the reaction solution is accommodated in the reaction container to which the feed discharged from the cotton for collection contracted by the descent of the first rod is accommodated.
The method according to claim 1,

The reaction inducing unit,

In a second rod having a lower end accommodated in the reaction vessel and an upper end coupled to a fixture of the measuring device, mounted on the lower side of the second rod, the sample discharged from the collecting cotton reacts with the reaction solution A side flow immunoassay kit for measuring multiple biomarkers in saliva, characterized in that it comprises a first impeller stirring the reaction solution to do so.
The method according to claim 1,

The reaction inducing unit,

a stopper capable of opening and closing the upper surface of the reaction vessel;

a second impeller mounted on the upper side of the stopper and agitating the reaction solution so that the sample discharged from the cotton for collection reacts with the reaction solution;

Further comprising a third rod having both upper and lower ends connected to the stopper and the bottom surface of the reaction vessel,

The stopper is, by a drive transmission unit including a docking station on which the reaction vessel is seated, rotating upward or downward with the reaction vessel while opening and closing the upper surface of the reaction vessel Measuring multiple biomarkers side flow immunoassay kit for
a measurement housing having an internal space that can accommodate a lateral flow immunoassay (LFA) cartridge comprising a polygonal column-shaped body and a reaction vessel accommodating a lower portion of the body and containing a reaction solution;

a drive transmission unit for generating a driving force for forward and reverse rotation of the docking station;

a CCD camera mounted on one side of the measuring housing to photograph a test line formed by the reaction solution reacting to the LFA strip provided on the lower side of the plurality of reaction surfaces provided in the vertical direction of the body; and

Measure multiple biomarkers in saliva with a side flow immunoassay kit comprising a processing unit that calculates the concentration of the target substance in the sample by comparing and analyzing the reaction determination data stored in advance from the test line photographed with the CCD camera A fully automatic measuring instrument.
8. The method of claim 7,

The drive transmission unit,

In the first rod having a lower end to which a cotton for collecting sample, including saliva, is collected in the oral cavity of the subject to be measured, the fixing member detachably coupled to the upper end of the first rod;

a joint head having a lower surface provided with the fixing member;

a first motor provided on the upper side of the measurement housing and connected to the joint head to rotate the joint head;

a second motor connected to the joint head to raise or lower the joint head;

A third motor provided on one side of the measurement housing and connected to the docking station to generate a driving force for rotating and stopping the polygonal column-shaped main body seated on the docking station by a predetermined angle, characterized in that it further comprises A fully automatic measuring device that measures multiple biomarkers in saliva with a lateral flow immunoassay kit.
8. The method of claim 7,

The drive transmission unit,

A second rod having a lower end accommodated in the reaction vessel and an upper end coupled to a fixing member of a measuring device, the second rod comprising: a fixing member detachably coupled to the upper end of the second rod;

a joint head having a lower surface provided with the fixing member;

a first motor provided on the upper side of the measurement housing and connected to the joint head to rotate the joint head;

a second motor connected to the joint head to raise or lower the joint head;

A third motor provided on one side of the measurement housing and connected to the docking station to generate a driving force for rotating and stopping the polygonal column-shaped main body seated on the docking station by a predetermined angle,

The first motor is a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that rotating a first impeller mounted on the lower side of the second rod.
8. The method of claim 7,

The drive transmission unit,

It is provided on one side of the measurement housing, is connected to the docking station, rotates and stops the polygonal column-shaped body seated in the docking station by a predetermined angle, and generates a driving force capable of forward and reverse rotation, and at the same time, the reaction vessel In the third rod extended to the stopper capable of opening and closing the upper surface of the third rod and accommodated in the reaction vessel, a fourth motor connected to the upper side of the third rod to rotate the third rod forward and reverse;

Further comprising a fifth motor for generating a driving force for raising or lowering the body seated in the docking station,

The fourth motor is a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that rotating the second impeller mounted on the upper side of the stopper.
9. The method of claim 8,

The predetermined angle is

The pattern of rotating and stopping by 360/n degrees so that n response surfaces of the main body in the shape of a polygonal column face the CCD camera provided in the measurement housing are repeated n-1 times or n times,

Wherein n is a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that it is a positive integer of 3 or more.
10. The method of claim 9,

The predetermined angle is

The pattern of rotating and stopping by 360/n degrees so that n response surfaces of the main body in the shape of a polygonal column face the CCD camera provided in the measurement housing are repeated n-1 times or n times,

Wherein n is a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that it is a positive integer of 3 or more.
11. The method of claim 10,

The predetermined angle is

The pattern of rotating and stopping by 360/n degrees so that the n response surfaces of the main body in the shape of a circle or polygonal column face the CCD camera provided in the measurement housing are repeated n-1 times or n times,

Wherein n is a fully automatic measuring device for measuring multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that it is a positive integer of 3 or more.
A first step of collecting a sample including saliva in the oral cavity of the subject with a collection cotton;

A second step of eluting the sample into the reaction solution in the LFA cartridge by putting the cotton for collection into the lateral flow immunoassay (LFA) cartridge seated in the measurement housing;

After the first time elapses, the cotton for collection is applied to the LFA cartridge so that the reaction solution from which the sample is eluted is reacted with a plurality of strips for LFA respectively formed on a plurality of reaction surfaces provided on the polygonal columnar body of the LFA cartridge. A third step of lowering further into the inside or opening the stopper formed in the reaction vessel containing the reaction solution to which the sample discharged from the collecting cotton reacts, and contacting the reaction solution with the plurality of strips for LFA;

After a second time period greater than the first time has elapsed, the LFA cartridge is rotated and stopped so that the plurality of reaction surfaces face the CCD camera provided in the measurement housing, and the control line and test line of the LFA strip are connected to the CCD camera. A fourth step of taking a picture; and

An image processor compares and analyzes the color development ratio between the control line and the test line with the reaction determination data stored in the image taken by the CCD camera to calculate the concentration of the target substance in the sample, and calculate the result of the calculation in the measurement housing A method of detecting a target substance using a fully automatic measuring device that measures multiple biomarkers in saliva with a side flow immunoassay kit, characterized in that it includes a fifth step of outputting it through a display panel formed on one side of the .