Lateral flow chromatography test strip comprising polycrystalline structure and manufacturing method thereof

Abstract

The invention relates to a lateral flow chromatography test strip comprising a polycrystalline structure and a manufacturing method thereof. More specifically, the present invention relates to a lateral flow chromatography test strip and a method for efficiently manufacturing the same, which can realize high-sensitivity field diagnosis by using a plasmon polycrystalline structure growth technique and can simultaneously detect a plurality of targets including polycrystalline structures, and more specifically, to a lateral flow chromatography test strip and a method for efficiently manufacturing the same.

Description

Technical Field

[0001] The present invention relates to a lateral flow chromatography test strip comprising a polycrystalline structure and a method for manufacturing the same. More specifically, the present invention relates to a lateral flow chromatography test strip comprising a polycrystalline structure and a method for efficiently manufacturing the same, which utilizes plasmonic polycrystalline structure growth technology to achieve high-sensitivity on-site diagnosis and simultaneously detect multiple targets. Background Art

[0002] The development of diagnostic methods and instruments that qualitatively or quantitatively measure trace amounts of substances such as nucleic acids and proteins contained in biological samples such as blood, urine, saliva, and tears continues to advance rapidly. Since the introduction of radioimmunoassay (RIA) using radioisotopes in the 1950s, enzyme immunoassays (ELISA) have been developed and refined in the 1970s and 1980s.

[0003] A representative method recently developed for detecting proteins or nucleic acids is the chromatography-based lateral flow assay (LFA). Lateral flow assays are a diagnostic method that typically uses antigen-antibody reactions on a membrane to visually confirm infection. This method exploits the phenomenon that as a sample, such as blood or urine, flows through a porous membrane via capillary flow, probes become immobilized on specific sites on the membrane through antigen-antibody reactions.

[0004] Conventional lateral flow chromatography test strips utilizing lateral flow chromatography consist of a sample pad for introducing the sample, a conjugate pad where the biochemical reaction between the sample and the probe occurs, an assay pad where the sample undergoes immunoassays as it flows through capillary action, and an absorbent pad downstream of the assay pad to absorb excess sample and maintain lateral flow. The assay pad also features a test line and a control line that immunoreact with the target.

[0005] This lateral flow chromatography method is widely used in many fields such as pregnancy diagnosis, cancer diagnosis, detection of the presence of other specific proteins or genes, or microbial detection.

[0006] However, conventional lateral flow chromatography (LFC) methods struggle to achieve high-sensitivity detection when the concentration of biological samples is low, making them difficult to use in point-of-care (POC) diagnostics. In particular, there is a lack of technology capable of highly sensitive detection of unlabeled biological samples.

[0007] Furthermore, conventional lateral flow test strips have difficulty in detecting multiple targets simultaneously and with high sensitivity.

[0008] Furthermore, when manufacturing lateral flow test strips using conventional methods, there are the following problems: forming multiple test lines, adjusting the distance between test lines, and forming test lines made of different materials are difficult.

[0009] As a background art of the present invention, Korean Patent Publication No. 10-2014-0110795 describes a biosensor test strip utilizing a redox cycle, wherein a product generated by a catalyst marker participates in the cycle. Summary of the Invention

[0010] Technical problem to be solved by the invention

[0011] An object of the present invention is to provide a lateral flow chromatography test strip that can diagnose label-free samples on-site with high sensitivity.

[0012] Another object of the present invention is to provide a lateral flow test strip that can simultaneously detect multiple targets with high sensitivity.

[0013] Another object of the present invention is to provide a method for manufacturing a lateral flow chromatography test strip, which is easy to form multiple test lines and test lines made of different materials, and easy to adjust the distance between the test lines.

[0014] The objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned should be clearly understood from the detailed description.

[0015] Means for solving technical problems

[0016] According to one aspect, a lateral flow chromatography test strip is provided, wherein the lateral flow chromatography test strip includes a test strip component, the test strip component includes a test area, the test area includes one or more detection portions where a biochemical reaction occurs between a target contained in the sample and a probe when the sample flows by capillary action, and the detection portion includes a polycrystalline structure having multiple grain boundaries composed of multiple nanoparticle clusters.

[0017] According to one embodiment, the test strip member may include, in a manner arranged in one direction, a sample area for introducing a sample; the test area; and an absorption area for absorbing excess sample to maintain lateral flow.

[0018] According to one embodiment, the lateral flow test strip can be used for surface enhanced Raman spectroscopy (SERS), fluorescence, or plasmon-enhanced fluorescence (PEF) analysis.

[0019] According to one embodiment, the nanoparticles may be composed of one or more of Au, Ag, and Pt.

[0020] According to one embodiment, for the lateral flow chromatography test strip, the polycrystalline structure may be combined with a gene probe.

[0021] According to one embodiment, the detection portion may include one or more of the following detection lines: i) two or more vertical detection lines perpendicular to the sample flow direction; ii) two or more horizontal detection lines horizontal to the sample flow direction; and iii) one or more vertical detection lines perpendicular to the sample flow direction and one or more horizontal detection lines horizontal to the sample flow direction.

[0022] According to one embodiment, the detection portion may include one or more vertical detection lines perpendicular to the sample flow direction and one or more circular, elliptical or polygonal detection portions located behind the vertical detection lines relative to the sample flow direction.

[0023] According to one embodiment, a hydrophobic treatment component that has been subjected to hydrophobic treatment may be provided locally around the detection portion in such a manner that samples of the vertical detection lines are collected at the detection portion.

[0024] According to one embodiment, the detection portion may further include a detection portion containing a foreign material.

[0025] According to one embodiment, the detection portion may include: a first detection portion, forming a graphene oxide layer, and two or more second detection portions, forming the polycrystalline structure; the first detection portion is combined with a gene probe labeled with a fluorescent molecule whose fluorescence is quenched, and the second detection portion is combined with a probe for binding to the fluorescent molecule, and the fluorescent molecule restores fluorescence when the gene probe binds to the miRNA target molecule. Thus, the lateral flow chromatography test strip of the present invention can be used as a multiplex sensor.

[0026] According to one embodiment, the probe attached to each of the second detection parts may be anti-FAM, anti-TAMRA, anti-methylene blue, anti-Cy5, anti-Cy3, anti-Alexa Fluor 488, anti-Rhodamine Red, anti-Texas Red, or anti-Fluorescein / Oregon Green.

[0027] According to one embodiment, the test strip component may be composed of one or more of cellulose acetate (CA), mixed cellulose ester (MCE), nitrocellulose (NC), CN95, and a filter membrane formed of a synthetic polymer.

[0028] According to one embodiment, the sample may be selected from one or more of cells, metabolites, proteins, nucleic acids, DNA, RNA, enzymes, organic molecules, viruses, extracellular vesicles, microvesicles, exosomes, and fat in urine, saliva, sweat, blood, and tears.

[0029] According to one embodiment, the detection portion may have a first detection portion and a second detection portion arranged in sequence relative to the flow direction of the sample, and the polycrystalline structure of the first detection portion that first contacts the sample is bound to a probe peptide dissociated by matrix metalloproteinase (MMP), and a probe for detecting the dissociated probe peptide is bound to the second detection portion, thereby the lateral flow chromatography test strip of the present invention can be used as an MMP sensor.

[0030] According to one embodiment, the detection section may have a first detection section relative to the sample flow direction and a second detection section and a third detection section located behind the first detection section, and the second detection section and the third detection section are parallel to each other. The polycrystalline structure of the first detection section is bound to a gene that reacts with an intercalation dye as a first control, the polycrystalline structure of the second detection section is bound to an antibody that can bind to methylated DNA (Me-DNA), and the polycrystalline structure of the third detection section is bound to the gene probe. Therefore, the lateral flow chromatography test strip of the present invention can be used as a Me-DNA sensor.

[0031] According to one embodiment, the detection portion may include a first detection portion, a second detection portion, and a third detection portion horizontally arranged relative to the sample flow direction, and the first detection portion, the second detection portion, and the third detection portion are parallel to each other, and the polycrystalline structure of the first detection portion is bound to a gene that reacts with an intercalation dye as a first control, the polycrystalline structure of the second detection portion is bound to an antibody that can bind to methylated DNA (Me-DNA), and the polycrystalline structure of the third detection portion is bound to the gene probe, so that the lateral flow chromatography test strip of the present invention can be used as a Me-DNA sensor.

[0032] According to another aspect, a method for manufacturing a lateral flow chromatography test strip is provided, comprising: a test strip component preparation step A-1) of preparing a test strip component for lateral flow chromatography; a tape application step A-2) of applying tape to areas of the test strip component other than areas to be patterned; a polycrystalline structure formation step A-3) of immersing the test strip component in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip component; and a tape removal step A-4) of removing the tape from the test strip component.

[0033] According to another aspect, a method for manufacturing a lateral flow chromatography test strip is provided, comprising: a test strip component preparation step B-1) of preparing a test strip component for lateral flow chromatography; a polycrystalline structure formation step B-2) of immersing the test strip component in a composition comprising a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip component; a tape application step B-3) of applying a tape to an area of the test strip component other than an area to be patterned; and a tape removal step B-4) of removing the tape from the test strip component.

[0034] According to another aspect, a method for manufacturing a lateral flow chromatography test strip is provided, comprising: a test strip member preparing step (C-1) of preparing a test strip member for lateral flow chromatography; a graphene oxide layer forming step (C-2) of forming a graphene oxide layer on the test strip member using a graphene oxide solution; a first tape attaching step (C-3) of attaching a tape to an area of the test strip member other than an area where graphene oxide patterning is to be performed; a first tape removing step (C-4) of removing the tape from the test strip member; a second tape attaching step (C-5) of attaching a tape to an area of the test strip member other than an area where a polycrystalline structure is to be patterned; a polycrystalline structure forming step (C-6) of immersing the test strip member in a composition comprising a noble metal precursor and a reducing agent solution to form a polycrystalline structure having multiple grain boundaries on the test strip member composed of a plurality of nanoparticle clusters; and a second tape removing step (C-7) of removing the tape from the test strip member.

[0035] According to another aspect, a method for manufacturing a lateral flow chromatography test strip is provided, comprising: a strip member preparing step D-1) preparing a strip member for lateral flow chromatography; a polycrystalline structure forming step D-2) immersing the strip member in a composition comprising a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the strip member; a first tape attaching step D-3) attaching a tape to an area of the strip member other than an area where the polycrystalline structure is to be patterned; a first tape removing step D-4) removing the tape from the strip member; a second tape attaching step D-5) attaching the tape to an area of the strip member other than an area where graphene oxide is to be patterned; a graphene oxide layer forming step D-6) forming a graphene oxide layer on the strip member using a graphene oxide solution; and a second tape removing step D-7) removing the tape from the strip member.

[0036] Effects of the Invention

[0037] According to one embodiment of the present invention, the lateral flow chromatography test strip of the present invention includes a plasmon polycrystalline structure, thereby enabling high-sensitivity on-site diagnosis of label-free biological samples.

[0038] According to one embodiment of the present invention, the lateral flow test strip of the present invention includes a plurality of detection lines including plasmon polycrystalline structures, thereby enabling high-sensitivity simultaneous detection of multiple targets.

[0039] According to one embodiment of the present invention, in the method for manufacturing a lateral flow chromatography test strip of the present invention, the polycrystalline structure is patterned by solution processing, which can easily generate multiple detection lines and detection lines of different materials, and can also effectively adjust the distance between detection lines. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 FIG1 is a diagram schematically illustrating a lateral flow chromatography test strip including a polycrystalline structure on a test line according to an embodiment of the present invention.

[0041] Figure 2 Part (a) shows a detection portion manufactured using cellulose acetate as a test strip member according to one embodiment of the present invention. Figure 2 Part (b) to Figure 2 Part (e) shows Figure 2 Scanning electron microscope (SEM) photograph of the detection part (a) Figure 2 (b) of the polycrystalline verification results ( Figure 2 (c) of the nanoparticles), the formation mechanism of nanoparticles ( Figure 2 (d) of the nanoparticles) and the nanoparticle distribution diagram ( Figure 2 (e) of the 2012 / 13 / EC meeting).

[0042] Figure 3 FIG2 is a diagram schematically illustrating a lateral flow chromatography test strip including polycrystalline structures in two or more detection portions of a test area according to an embodiment of the present invention. Figure 3 Part (a) shows a test strip comprising multiple detection sections, Figure 3 Part (b) shows a test strip including multiple detection sections with adjustable spacing, Figure 3 Part (c) shows a test strip comprising multiple polycrystalline structures, Figure 3 Part (d) shows a test bar including a polycrystalline structure made with a variety of test bar components, Figure 3 Part (e) shows a test strip including a vertical detection portion and a circular detection ring.

[0043] Figure 4 The graph shows the results of measuring SERS signals of methylene blue at different concentrations using a lateral flow chromatography test strip including a polycrystalline structure in a detection line according to an embodiment of the present invention.

[0044] Figure 5 This is an image showing a SERS signal uniformly displayed only in the polycrystalline structure formation region by plasmon mapping of a detection line of a lateral flow chromatography test strip including a polycrystalline structure in the detection line according to an embodiment of the present invention.

[0045] Figure 6 The figures show the fluorescence signals of methylene blue at different concentrations using a lateral flow chromatography test strip including a polycrystalline structure in a detection line according to an embodiment of the present invention.

[0046] Figure 7a Figure 2 is a photograph of a lateral flow test strip including a detection portion of a heterogeneous material and a scanning electron microscope (SEM) image of each detection line according to an embodiment of the present invention. The heterogeneous material detection portion includes a detection line including a polycrystalline structure and a detection line including graphene oxide.

[0047] Figure 7b FIG2 is a diagram illustrating SERS signals and fluorescence signals measured using a lateral flow chromatography test strip having heterogeneous materials according to an embodiment of the present invention. The heterogeneous materials include a detection line having a polycrystalline structure and a detection line having a graphene oxide layer.

[0048] Figure 8 The figure schematically shows a lateral flow chromatography test strip including a polycrystalline structure used as a matrix metalloproteinase (MMP) sensor according to one embodiment.

[0049] Figure 9Part (a) shows an image of Raman mapping performed on a vertical detection line of a polycrystalline structure according to one embodiment of the present invention, namely, an MMP probe peptide in the first detection portion, which was immobilized at different concentrations.

[0050] Figure 9 Part (b) to Figure 9 Part (d) is a graph showing Raman spectra of the vertical detection line of the polycrystalline structure according to one embodiment of the present invention, i.e., the first detection portion, where the MMP probe peptide is plotted at different concentrations, a calibration curve, and signal uniformity in the first detection portion.

[0051] Figure 10 Part (a) is an image of Raman mapping performed on the second detection part according to different concentrations of Tamra probe. The second detection part is a detection circle located behind the first detection part at a predetermined distance according to one embodiment of the present invention.

[0052] Figure 10 Part (b) to Figure 10 Part (d) shows Raman spectra of the detection zone of the polycrystalline structure according to one embodiment of the present invention, that is, the Tamra probe of the second detection part, which are plotted at different concentrations, a calibration curve, and a graph of signal uniformity in the second detection part area.

[0053] Figure 11 1 is a comparative image of the intensity of the Raman signal detected under different designs of the detection zone of the polycrystalline structure, ie, the hydrophobic treatment area around the second detection portion according to an embodiment of the present invention.

[0054] Figure 12 Part (a) is a photograph showing a lateral flow test strip including a vertical detection line (ie, a first detection portion) and a detection circle (ie, a second detection portion) of a polycrystalline structure according to one embodiment of the present invention.

[0055] Figure 12 Part (b) and Figure 12 Part (c) is to pass the sample containing MMP through Figure 12 Part (a) shows Raman mapping images of a lateral flow test strip before and after the first detection section.

[0056] Figure 12 Part (d) is to pass the sample containing MMP through Figure 12 Part (a) shows a Raman mapping image of the second detection section after the lateral flow test strip.

[0057] Figure 13 FIG1 is a schematic diagram schematically illustrating a method of using a lateral flow chromatography test strip including a polycrystalline structure as a methylated DNA (Me-DNA) sensor according to an embodiment.

[0058] Figure 14 Part (a) of FIG. 1 is an image showing fluorescence signal measurement results corresponding to different intercalating dyes when a lateral flow chromatography test strip including a polycrystalline structure according to an embodiment is used as a methylated DNA (Me-DNA) sensor.

[0059] Figure 14 Part (b) is an image showing Raman mapping results corresponding to different intercalating dyes when a lateral flow chromatography test strip including a polycrystalline structure according to an embodiment of the present invention is used as a methylated DNA (Me-DNA) sensor.

[0060] Figure 15 FIG1 is a schematic diagram showing a method of using a lateral flow chromatography test strip comprising a polycrystalline structure according to an embodiment of the present invention as a miRNA sensor.

[0061] Figure 16 To schematically illustrate a method (Method A), a lateral flow chromatography test strip comprising a polycrystalline structure is manufactured by patterning to form a polycrystalline structure according to one embodiment of the present invention.

[0062] Figure 17 Comparative images showing the shapes of polycrystalline structures formed by different types of reducing agents when CA is used as a test strip component when a lateral flow chromatography test strip including a polycrystalline structure is manufactured using a method for manufacturing a polycrystalline structure (method A) according to an embodiment of the present invention.

[0063] Figure 18 Comparative images showing the shapes of polycrystalline structures formed from different types of strip components when manufactured using the same reducing agent using a method for manufacturing a lateral flow chromatography test strip including a polycrystalline structure according to an embodiment of the present invention (Method A).

[0064] Figure 19 To schematically illustrate a method (Method B), a polycrystalline structure is formed according to one embodiment of the present invention and then patterned to manufacture a lateral flow chromatography test strip including the polycrystalline structure.

[0065] Figure 20 Comparative images showing the shapes of polycrystalline structures formed using different types of reducing agents in a method (method B) of forming a polycrystalline structure and then patterning the structure to manufacture a lateral flow chromatography test strip comprising the polycrystalline structure according to an embodiment of the present invention.

[0066] Figure 21 1 is a comparative image showing the height of the detection portion of a test strip component formed according to different reducing agent types and reducing agent mixing ratios in a method (method B) of forming a polycrystalline structure and then patterning the polycrystalline structure to manufacture a lateral flow chromatography test strip including the polycrystalline structure according to one embodiment of the present invention.

[0067] Figure 22 The present invention is schematically illustrated as a method (method C) for manufacturing a lateral flow chromatography test strip including a heterogeneous material detection portion according to an embodiment of the present invention, wherein the heterogeneous material detection portion has a detection line including a polycrystalline structure (polycrystalline nanostructure detection portion) and a detection line including graphene oxide (graphene oxide detection portion).

[0068] Figure 23 Part (a) is based on Figure 22 Scanning electron microscopy (SEM) image of the polycrystalline nanostructured detection portion of the fabricated lateral flow test strip.

[0069] Figure 23 Part (b) is based on Figure 22 Scanning electron microscopy (SEM) image of the graphene oxide detection portion of the fabricated lateral flow chromatography test strip. DETAILED DESCRIPTION

[0070] The objectives, specific advantages and novel features of the present invention will become more apparent from the following detailed description and examples with reference to the accompanying drawings.

[0071] Previously, the terms or words used in this specification and claims should not be interpreted as having a general and dictionary meaning, but should be interpreted as the meaning and concept that is consistent with the technical idea of the present invention from the perspective that the inventor can reasonably define the terminology in order to best explain his or her invention.

[0072] In the specification, when a component such as a layer, part, or substrate is described as being "on," "connected to," or "coupled to" another component, this may mean that it is directly "on," "connected to," or "coupled to" the other component, and may also mean that one or more other components may be present between the two components. Conversely, when a component is described as being "directly on," "directly connected to," or "directly coupled to" another component, no other components may be present between the two components.

[0073] The terms used in this specification are only used to describe specific embodiments and are not intended to limit the present invention. A singular quantity expression includes a plural quantity expression unless the context clearly excludes it.

[0074] In this specification, terms such as "include" or "have" should be understood as intending to specify the existence of features, numbers, steps, operations, constituent elements, parts or their combinations, without excluding in advance the existence or additional possibilities of one or more other features, numbers, operations, constituent elements, parts or their combinations.

[0075] In this specification, when a portion is referred to as "including" a certain component, this means that other components may be included, and does not exclude other components, unless explicitly stated otherwise. Furthermore, throughout this specification, the term "on" refers to a state of being above or below the subject portion, and does not necessarily mean being above the subject portion relative to the direction of gravity.

[0076] The present invention is susceptible to numerous variations and embodiments. Therefore, specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this does not limit the present invention to these specific embodiments, but rather should be understood to encompass all variations, equivalents, and alternatives within the spirit and technical scope of the present invention. When describing the present invention, if a detailed description of a related known art is determined to obscure the main purpose of the present invention, such detailed description will be omitted.

[0077] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. When describing with reference to the accompanying drawings, identical or corresponding components will be denoted by the same reference numerals, and redundant descriptions thereof will be omitted.

[0078] Figure 1 FIG1 is a diagram schematically illustrating a lateral flow chromatography test strip including a polycrystalline structure on a test line according to an embodiment of the present invention.

[0079] Reference Figure 1 A lateral flow chromatography test strip 1 according to one aspect of the present invention is composed of a sample area 10, a test area 20, and an absorption area 30 arranged in one direction.

[0080] The sample area 10 is an area for introducing samples, and may be configured to have a groove for accommodating a quantitative sample.

[0081] The test area 20 may be located downstream of the sample area 10 and include one or more detection sections (a first detection section 22 and a second detection section 24). As the sample flows, a biochemical reaction occurs between the target contained in the sample and the probe. The biochemical reaction is not particularly limited as long as the target can be detected, and includes immune reactions, chemical reactions (enzyme reactions, DNA reactions), and the like.

[0082] In the present invention, the detection portion (the first detection portion 22 and the second detection portion 24 ) is characterized by including a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries.

[0083] The absorption zone 30 is located downstream of the testing zone 20 and is configured to maintain lateral flow by absorbing excess sample.

[0084] Figure 2Part (a) shows a detection portion manufactured using cellulose acetate as a test strip member according to one embodiment of the present invention. Figure 2 Part (b) to Figure 2 Part (e) shows Figure 2 Scanning electron microscope (SEM) photograph of the detection part (a) Figure 2 (b) of the polycrystalline verification results ( Figure 2 (c) of the nanoparticles), the formation mechanism of nanoparticles ( Figure 2 (d) of the nanoparticles) and the nanoparticle distribution diagram ( Figure 2 (e) of the 2012 / 13 / EC meeting). Figure 3 FIG2 is a diagram schematically illustrating a lateral flow chromatography test strip including polycrystalline structures in two or more detection portions of a test area according to an embodiment of the present invention. Figure 3 Part (a) shows a test strip comprising multiple detection sections, Figure 3 Part (b) shows a test strip including multiple detection sections with adjustable spacing, Figure 3 Part (c) shows a test strip comprising multiple polycrystalline structures, Figure 3 Part (d) shows a test bar including a polycrystalline structure made with a variety of test bar components, Figure 3 Part (e) shows a test strip including a vertical detection portion and a circular detection ring.

[0085] Reference Figure 2 Part (a) to Figure 2 Part (e) and Figure 3 , it can be confirmed that cellulose acetate (see Figure 2 Part (a) to Figure 2 (e) of the present invention) and a polycrystalline structure composed of nanoparticle clusters formed on various test strip components other than these (see Figure 3 (d) of the 2015 / 16 / EC meeting).

[0086] Furthermore, refer to Figure 2 (d), although not limited thereto, shows that a gold (Au) precursor in a noble metal forms nanoparticles and aggregates into clusters, and in particular, it is grown directly on a test strip member using a solution process. Specifically, the aggregated nanoparticles form small-sized polygonal structure grains through coalescence, and then the particles gradually grow significantly through oriented attachment growth. Thus, a polycrystalline structure with multiple grain boundaries (GB) is formed. Although not limited thereto, the polycrystalline structure, i.e., the noble metal coral-like particles, can grow to an average size of micrometers ( Figure 2(e) of the present invention). For the polycrystalline structure having multiple grain boundaries as described above, since scattering is enhanced at the multiple grain boundaries, the signal intensity and signal uniformity can be significantly improved when used as a lateral flow chromatography test strip.

[0087] Although not limited thereto, the nanoparticles are connected by branch structures, and the average diameter of the polycrystalline structure can be 1 μm to 100 μm. The polycrystalline structure of the detection portion of the lateral flow chromatography test strip of the present invention can be grown by solution processing and connected by branch structures, and can have an average diameter of 0.1 μm to 100 μm. Although not limited thereto, the average diameter of the polycrystalline structure within this range is suitable for improving signal strength and signal uniformity.

[0088] There is no particular limitation on the test strip components applicable to the present invention, and a variety of materials can be used. Figure 3 In part (d), the test strip component may be composed of one or more of cellulose acetate (CA), mixed cellulose ester (MCE), nitrocellulose (NC), CN95, and a filter membrane formed of a synthetic polymer. Although not limited thereto, the filter membrane may be composed of polyethersulfone, polycarbonate, glass fiber, nylon, or polytetrafluoroethylene.

[0089] Reference Figure 3 Although not limited thereto, the first detection unit 22 and the second detection unit 24 may include one or more of the following:

[0090] i) Two or more vertical detection lines perpendicular to the sample flow direction (see Figure 3 (a) of the Regulations);

[0091] ii) Two or more horizontal detection lines parallel to the direction of sample flow (see Figure 3 (c) of the Regulations);

[0092] iii) one or more vertical test lines perpendicular to the sample flow direction and one or more horizontal test lines parallel to the sample flow direction (see Figure 3 (c) of the 2015 / 2016 / EC).

[0093] As described above, there may be multiple combinations of one or more vertical detection lines and one or more horizontal detection lines.

[0094] Furthermore, the detection portion may include one or more first detection portions 22 (vertical detection lines perpendicular to the sample flow direction) and one or more second detection portions 24 (circular detection portions located behind the vertical detection lines, i.e., the first detection portions 22, relative to the sample flow direction) (see Figure 3 That is, there may be various combinations, such as one vertical detection line, i.e., the first detection unit, and two or more detection circles, i.e., the second detection unit, or two or more vertical detection lines, i.e., the first detection unit, and one detection circle, i.e., the second detection unit.

[0095] The shape of the detection ring of the second detection portion 24 is not particularly limited as long as it can further concentrate the trace sample. Therefore, although not limited thereto, in addition to being circular, the detection ring can also be deformed into an elliptical or polygonal shape.

[0096] Although not limited thereto, a hydrophobic treatment area 26 may be locally provided around the detection circle, i.e., the second detection section 24, so that the sample passing through the vertical detection line, i.e., the first detection section 22, is collected in the detection circle, i.e., the second detection section 24. Although not limited thereto, to enhance the signal, the hydrophobic treatment area 26 preferably minimizes the sample flow area between the vertical detection line, i.e., the first detection section 22, and the detection circle, i.e., the second detection section 24, while maintaining the portion of the sample flow area in contact with the detection circle, i.e., the second detection section 24 occupying approximately 1 / 3 to 1 / 4 of the circumference of the detection circle, i.e., the second detection section 24 (see Figure 3 (e) of the present invention). In addition, the hydrophobic treatment can utilize various well-known techniques. Although not limited thereto, the hydrophobic treatment can be easily achieved using a hydrophobic pen.

[0097] Figure 4 The graph shows the results of measuring SERS signals of methylene blue at different concentrations using a lateral flow chromatography test strip including a polycrystalline structure in a detection line according to an embodiment of the present invention. Figure 5 This is an image showing a SERS signal uniformly displayed only in the polycrystalline structure formation region by plasmon mapping of a detection line of a lateral flow chromatography test strip including a polycrystalline structure in the detection line according to an embodiment of the present invention. Figure 6 The figures show the fluorescence signals of methylene blue at different concentrations using a lateral flow chromatography test strip including a polycrystalline structure in a detection line according to an embodiment of the present invention.

[0098] Reference Figures 4 to 6Although not limited thereto, the lateral flow chromatography test strip of the present invention can be used for surface enhanced Raman spectroscopy (SERS), fluorescence or plasmon-enhanced fluorescence (PEF) analysis. The lateral flow chromatography test strip of the present invention includes a plasmon polycrystalline structure to enable highly sensitive and accurate on-site diagnosis when analyzing unlabeled biological samples by SERS, fluorescence or PEF. In particular, Figure 4 and Figure 5 As shown in Figure 2, at wavelengths of 633 nm and 785 nm, SERS signals can be used to achieve highly reliable quantitative analysis. Figure 6 As shown, quantitative analysis can be achieved using PEF. Although not limited thereto, depending on the type of test strip component, SERS or fluorescence analysis may be more suitable. For example, CA may be more suitable for SERS analysis, while NC may be more suitable for fluorescence analysis.

[0099] Figure 7a Figure 2 is a photograph of a lateral flow test strip including a detection portion of a heterogeneous material and a scanning electron microscope (SEM) image of each detection line according to an embodiment of the present invention. The heterogeneous material detection portion includes a detection line including a polycrystalline structure and a detection line including graphene oxide. Figure 7b 1 is a diagram illustrating SERS signals and fluorescence signals measured using a lateral flow chromatography test strip having heterogeneous materials according to an embodiment of the present invention. The heterogeneous material includes a detection line comprising a polycrystalline structure, namely the first detection portion 22, and a detection line comprising a graphene oxide layer, namely the second detection portion 24.

[0100] like Figure 7a and Figure 7b As shown, the first detection section 22 and the second detection section 24 may include a detection line, and the detection line may include a heterogeneous material. The heterogeneous material may be composed of other materials that are not polycrystalline, and there is no particular limitation as long as the material is a control material for accurately detecting the target or a material that can simultaneously detect multiple targets.

[0101] For example, Figure 7a and Figure 7b A lateral flow chromatography test strip is shown, comprising a detection line of a graphene oxide layer (GO line) as a first detection portion 22 and a detection line comprising a polycrystalline structure (Au line) as a second detection portion 24, thereby being capable of being used as a multiplex sensor.

[0102] like Figure 7b As shown in the left graph of , in the detection line (Au line) including the polycrystalline structure, ie, the second detection section 24 , the sample can be analyzed by the SERS signal.

[0103] Furthermore, although not limited thereto, the detection line (GO line) comprising a graphene oxide layer, i.e., the first detection portion 22, can be combined with a gene probe labeled with a fluorescent molecule, and the detection line (Au line) comprising a polycrystalline structure, i.e., the second detection portion 24, can be combined with an antibody that can bind to the fluorescent molecule.

[0104] Therefore, if Figure 7b As shown in the right figure, when the sample flows, the fluorescent molecule that binds to the gene probe in the first detection section 22 (the detection line (GO line) including the graphene oxide layer) and thus quenches the fluorescence, when the gene probe binds to the miRNA target molecule, etc., thereby dissociating from the gene probe, the fluorescence is restored. In the detection line (Au line) including the polycrystalline structure, i.e., the second detection section 24, the fluorescent molecule binds to the antibody that can bind to the fluorescent molecule, thereby enabling the detection of a fluorescent signal. For a detailed mechanism of fluorescence recovery, see "Angew. Chem Int. Ed, 48(26)(2009). pp4785".

[0105] Although not limited thereto, the antibody attached to each of the second detection portions 24 may be anti-FAM, anti-TAMRA, anti-methylene blue, anti-Cy5, anti-Cy3, anti-Alexa Fluor 488, anti-Rhodamine Red, anti-Texas Red, or anti-Fluorescein / Oregon Green.

[0106] Although not limited thereto, the sample may be selected from one or more of urine, saliva, sweat, blood, and tears. One or more of cells, metabolites, proteins, nucleic acids, DNA, RNA, enzymes, organic molecules, viruses, extracellular vesicles, microvesicles, exosomes, and fat may be detected in the sample.

[0107] Figure 8 The figure schematically shows a lateral flow chromatography test strip including a polycrystalline structure used as a matrix metalloproteinase (MMP) sensor according to one embodiment.

[0108] Reference Figure 8 The detection portion has a vertical detection line, namely a first detection portion 22, and a detection circle, namely a second detection portion 24, which are perpendicular to the flow direction of samples such as tears and blood. The polycrystalline structure of the first detection portion 22 that first contacts the sample is bound to a probe peptide dissociated by MMP, and the detection circle, namely the second detection portion 24, which is located behind the first detection portion 22 relative to the sample flow direction, is bound to a probe for detecting the dissociated probe peptide. Therefore, the lateral flow chromatography test strip of the present invention can be used as an MMP sensor.

[0109] MMPs, as cancer biomarkers, can induce immune responses, but are present in blood, saliva, and urine at concentrations of only ng / mL, making them difficult to detect. According to the above configuration, as the sample flows, the probe peptide bound to the polycrystalline structure in the first detection section 22 is dissociated by the MMP. The dissociated probe peptide then binds to the polycrystalline structure in the second detection section 24, enabling detection via SERS or PEF analysis. Furthermore, the probe peptide binds to an antibody, a probe used to detect the probe peptide bound to the polycrystalline structure, enabling detection via SERS or PEF analysis.

[0110] Although not limited thereto, in the first detection portion 22 , the binding of the polycrystalline structure to the probe peptide may be achieved via a dye-labeled peptide-SH, the sequence of which is: Mca-Lys-Pro-Leu-Gly-Leu-Dap(Dnp)-Ala-Arg.

[0111] Although not limited thereto, the antibody that binds to the probe peptide in the second detection section 24 may be an anti-Tamra antibody.

[0112] In the present invention, various known techniques can be used for antibody attachment and dye-labeled probe peptide dissociation by MMP. For reference, see "Fransiska SHKrismastuti, Stephanie Pace and Nicolas H. Voelcker, Adv. Funct. Mater. 2014, 24, 3639-3650 and Ying Ye, Yuancai Ge, Qingwen Zhang, Meiling Yuan, Yu Cai, Kang Li, Yang Li, Ruifeng Xie, Changshun Xu, Danfeng Jiang, Jia Qu, Xiaohu Liu, and Yi Wang Ying Ye, Adv. Sci. 2022, 2104738."

[0113] Figure 9 Part (a) shows an image in which the MMP probe peptide in the vertical detection line of the polycrystalline structure according to one embodiment of the present invention, ie, the first detection portion 22, is fixed at different concentrations and Raman mapping is performed. Figure 9 Part (b) to Figure 9 Part (d) is a graph showing Raman spectra, calibration curves, and signal uniformity in the first detection section 22 region of the vertical detection line of the polycrystalline structure according to one embodiment of the present invention, i.e., the MMP probe peptide in the first detection section 22, plotted at different concentrations.

[0114] Reference Figure 9 Part (a) to Figure 9 In part (d), SERS analysis confirmed that the Raman signal increased in a manner dependent on the concentration of the MMP-dissociated probe peptide. Furthermore, although not limited to this, the MMP-dissociated probe peptide could be detected even at concentrations as low as 0.5 μM, with high signal uniformity, ensuring the reliability of the measurement results.

[0115] Figure 10 Part (a) is an image of Raman mapping performed on the second detection part according to different concentrations of Tamra probe, where the second detection part is a detection circle located behind the first detection part at a specified distance according to one embodiment of the present invention. Figure 10 Part (b) to Figure 10 Part (d) shows the Raman spectra of the Tamra probe of the detection zone of the polycrystalline structure according to one embodiment of the present invention, that is, the second detection part 24, which are plotted at different concentrations, a calibration curve, and a graph of the signal uniformity in the second detection part 24 area.

[0116] Reference Figure 10 Part (a) to Figure 10 In part (d), SERS analysis confirmed that the Raman signal increased in a manner dependent on the concentration of the Tamra probe peptide. Furthermore, while not limited to this, the Tamra probe can be detected even at concentrations as low as approximately 10 nM, with high signal uniformity, ensuring the reliability of the measurement results.

[0117] As mentioned above, refer to Figure 9 and Figure 10 , which can be used as an MMP sensor with excellent sensitivity and reliability.

[0118] Figure 11 1 is a comparative image of the intensity of the Raman signal detected under different designs of the detection circle of the polycrystalline structure, ie, the hydrophobic treatment area 26 around the second detection portion 24 according to one embodiment of the present invention.

[0119] Reference Figure 11 , although not limited thereto, it has been confirmed that by locally providing a hydrophobic treatment area 26 around the detection circle, i.e., the second detection part 24, so that the sample passing through the vertical detection line, i.e., the first detection part 22, is collected to the detection circle, i.e., the second detection part 24, the signal can be effectively enhanced. Although not limited thereto, the hydrophobic treatment area 26 preferably minimizes the sample flow area between the vertical detection line, i.e., the first detection part 22, and the detection circle, i.e., the second detection part 24 (see Figure 11 Part (a) to Figure 11(c) of the portion). In addition, the portion where the sample flows through the region and the detection circle, i.e., the second detection portion 24, contacts about 1 / 3 to 1 / 4 of the circumference of the detection circle, i.e., the second detection portion 24 (see Figure 11 Part (c) to Figure 11 Part (d) of the embodiment is suitable for enhancing the signal.

[0120] Figure 12 Part (a) is a photograph showing a lateral flow test strip including a vertical detection line, ie, a first detection portion 22, and a detection circle, ie, a second detection portion 24, of a polycrystalline structure according to an embodiment of the present invention. Figure 12 Part (b) and Figure 12 Part (c) is the sample containing MMP flowing through Figure 12 Part (a) shows Raman mapping images of the first detection portion 22 before and after the lateral flow test strip.

[0121] Reference Figure 12 Part (b) to Figure 12 In part (c), it can be confirmed that the SERS signal intensity of the MMP-containing sample after passing through the first detection part 22 is lower than that before passing through. Therefore, MMP can be detected based on the attenuation of the SERS signal caused by the change in the probe peptide concentration due to the dissociation of MMP. Figure 12 In part (d), MMP can be detected based on the enhanced SERS signal of the second detection unit 24 .

[0122] Figure 13 FIG1 is a schematic diagram schematically illustrating a method of using a lateral flow chromatography test strip including a polycrystalline structure as a methylated DNA (Me-DNA) sensor according to an embodiment.

[0123] Reference Figure 13 , the detection part may include: a first detection part 22, a vertical detection line perpendicular to the flow direction of samples such as blood, saliva, and urine; a second detection part 24a, a first horizontal detection line horizontally arranged behind the vertical detection line, i.e., the first detection part, relative to the flow direction of the sample; and a third detection part 24b, which is a second horizontal detection line. The second detection part 24a and the third detection part 24b may be parallel to each other. The polycrystalline structure of the vertical detection line, i.e., the first detection part 22, is combined with a gene that reacts with an intercalation dye as a first control, the polycrystalline structure of the first horizontal detection line, i.e., the second detection part 24, may be combined with an antibody that binds to methylated DNA (Me-DNA), and the polycrystalline structure of the second horizontal detection line, i.e., the third detection part 24b, is combined with the gene probe, thereby the lateral flow chromatography test strip of the present invention can be used as a Me-DNA sensor.

[0124] Me-DNA, as a cancer biomarker, exists in blood, saliva, and urine at levels of fM to nM, making it difficult to detect.

[0125] Although not limited thereto, the gene may be dsDNA, dsRNA, aptamer, ssDNA, or ssRNA.

[0126] The intercalating dye includes any dye that can intercalate into genes, and includes, for example, methylene blue, SYBR Green, EVA Green, ROX dye, Thiazole Red, TOTO, and VeriFluor far-red dye.

[0127] In the present invention, the gene probe or probe is not particularly limited as long as it can confirm the presence of a gene or other substance. Although not limited thereto, the gene probe may be an antibody, dye, etc. that binds to or reacts with the gene.

[0128] Reference Figure 13 In section (b), the vertical detection line, or first detection section 22, serves as a first control, incorporating a gene that reacts with an intercalating dye. The first horizontal detection line, or second detection section 24a, is incorporating an antibody that binds to Me-DNA. The second horizontal detection line, or third detection section 24b, serves as a second control for dsDNA binding and is incorporating an antibody that binds to dsDNA.

[0129] Reference Figure 13 Part (b) and Figure 13 In part (c), after the sample is flowed in the first detection section 22 as the first vertical detection line and the first control, MB or SYBR Green dye is injected to generate a reaction. Figure 13 In part (c), the first detection portion 22 is marked in black as a mark indicating that a reaction has occurred.

[0130] Then, if the sample contains Me-DNA, it can be bound to the Me-DNA antibody present in the first horizontal detection line, that is, the second detection part 24a, and detected by SERS or PEF. In this case, the second detection part 24a is marked in black, and if it is not detected, it is marked in gray. In addition, a reaction also occurs in the third detection part 24b, which serves as the second horizontal detection line and the second control and is bound to the dsDNA antibody, and the third detection part 24b is also marked in black. In this case, it can be marked as follows Figure 13 As shown in the lower right schematic diagram of part (c), it was determined to be Me-DNA positive.

[0131] On the other hand, if the first horizontal detection line, that is, the second detection portion 24a, and the second horizontal detection line, that is, the third detection portion 24b, do not react, Figure 13 As shown in the upper left schematic diagram of part (c), it can be determined as negative. Moreover, even if a reaction occurs only in the second horizontal detection line, that is, the second control, that is, the third detection part 24b, and no reaction is confirmed in the first horizontal detection line, that is, the second detection part 24a ( Figure 13 In the case of the lower left schematic diagram of part (c), it can also be determined as Me-DNA negative. If a reaction is confirmed in the second horizontal detection line, that is, the second detection part 24a, and no reaction occurs in the second horizontal detection line, that is, the second control, that is, the third detection part 24b ( Figure 13 In the case of the upper right schematic diagram in part (c), it can be determined as a false positive.

[0132] Reference Figure 13 In part (d), the design of the detection line can be diversified. Figure 13 As shown in part (d), the detection portion has a first horizontal detection line, i.e., a first detection portion 22, a second horizontal detection line, i.e., a second detection portion 24a, and a third horizontal detection line, i.e., a third detection portion 24b, which are horizontally arranged relative to the sample flow direction, and the first detection portion, the second detection portion, and the third detection portion can be parallel to each other.

[0133] The polycrystalline structure of the first horizontal detection line, i.e., the first detection section 22, is bound to a gene for an intercalation dye reaction as a first control, the polycrystalline structure of the second horizontal detection line, i.e., the second detection section 24a, is bound to an antibody that can bind to Me-DNA, and the polycrystalline structure of the third horizontal detection line, i.e., the third detection section 24b, is bound to the gene probe. Thus, the lateral flow chromatography test strip of the present invention can be used as a Me-DNA sensor.

[0134] Figure 14 Part (a) of FIG. 1 is an image showing fluorescence signal measurement results corresponding to different intercalating dyes when a lateral flow chromatography test strip including a polycrystalline structure according to an embodiment is used as a methylated DNA (Me-DNA) sensor. Figure 14 Part (b) is an image showing Raman mapping results corresponding to different intercalating dyes when a lateral flow chromatography test strip including a polycrystalline structure according to an embodiment of the present invention is used as a methylated DNA (Me-DNA) sensor.

[0135] like Figure 14As shown in part (a), the degree of fluorescence signal generation varies depending on the dye type, but overall, it can be confirmed that the fluorescence signal intensity is higher when DNA is present than when DNA is absent. In particular, the areas selected with black squares correspond to dyes that show a significant fluorescence signal enhancement effect.

[0136] Figure 14 Part (b) shows the extent of Raman signal formation for different dye types in response to the presence or absence of DNA. The areas highlighted by black squares correspond to dyes whose Raman signals are reduced in the presence of DNA.

[0137] Therefore, if Figure 14 Part (a) to Figure 14 As shown in part (b) of the figure, the degree of fluorescence enhancement or Raman signal reduction varies depending on the dye type. However, quantitative evaluation can still be performed using the trend of fluorescence enhancement or Raman signal reduction.

[0138] Figure 15 FIG1 is a schematic diagram showing a method of using a lateral flow chromatography test strip comprising a polycrystalline structure according to an embodiment of the present invention as a miRNA sensor.

[0139] Reference Figure 15 The detection part includes a first detection line, i.e., a first detection part 22, which forms a graphene oxide layer, and two or more second detection lines, i.e., second detection parts 24, which form the polycrystalline structure. The first detection line, i.e., the first detection part 22, is combined with a gene probe labeled with a fluorescent molecule with fluorescence quenching, and the second detection lines, i.e., the second detection parts 24a, 24b, and 24c, are combined with antibodies that can bind to the fluorescent molecules. Therefore, the lateral flow chromatography test strip of the present invention can be used as a multiplex sensor.

[0140] As cancer biomarkers, miRNAs are present in the pM to nM range or ng / mL in blood, urine, and other tissues, making them difficult to detect. In the present invention, a fluorescent molecule-labeled gene probe refers to a gene that has been conjugated to a fluorescent molecule. Examples of such gene probes include RNA probes, DNA probes, and aptamers.

[0141] Reference Figure 15 For the fluorescent molecules that are bound to the gene probe in the first detection line, i.e., the first detection part 22, and thus have their fluorescence quenched when the samples such as blood and urine flow, when the gene probe is bound to the miRNA target molecule and thus dissociated from the gene probe, the fluorescence is restored, and in the second detection part, i.e., the second detection part 24, it is bound to the antibody that can bind to the fluorescent molecule, thereby enabling the detection of the fluorescent signal.

[0142] As described above, the present invention can be used as a multiplex sensor because multiple genes can be detected simultaneously in two or more second detection lines, ie, second detection sections 24. Conventional methods cannot immobilize antibodies at different positions on one line.

[0143] Although not limited thereto, the antibody attached to each of the second detection lines, i.e., the second detection portion, may be anti-FAM, anti-TAMRA, anti-methylene blue, anti-Cy5, anti-Cy3, anti-Alexa Fluor 488, anti-Rhodamine Red, anti-Texas Red, or anti-Fluorescein / Oregon Green.

[0144] Figure 16 To schematically illustrate a method (Method A), a lateral flow chromatography test strip comprising a polycrystalline structure is manufactured by patterning to form a polycrystalline structure according to one embodiment of the present invention.

[0145] like Figure 16 According to another aspect of the present invention, a method for manufacturing a lateral flow chromatography test strip (method A) includes: a test strip member preparation step A-1) of preparing a test strip member 40 for lateral flow chromatography; a tape attachment step A-2) of attaching a tape 42 to an area of the test strip member 40 other than an area to be patterned; a polycrystalline structure formation step A-3) of immersing the test strip member 40 in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member 40; and a tape removal step A-4) of removing the tape 42 from the test strip member 40.

[0146] In such Figure 16 In the test strip member preparation step (A-1) shown in part (a), the test strip member 40 can be prepared from one or more of cellulose acetate (CA), mixed cellulose ester (MCE), nitrocellulose (NC), CN95, and a filter membrane formed from a synthetic polymer. Although not limited thereto, the filter membrane can be prepared from polyethersulfone, polycarbonate, glass fiber, nylon, or polytetrafluoroethylene, for example.

[0147] In such Figure 16In the tape attaching step (A-2) shown in part (b), the tape 42 is attached to the surface of the test strip member 40 excluding the area to be patterned. That is, the tape 42 is attached to the area excluding the area where the detection portion including the polycrystalline structure is formed. In this case, although not limited to this, any polymer-based tape containing an adhesive component can be used, such as polyimide tape, transparent tape (ordinary), or sticky note tape.

[0148] In such Figure 16 In the polycrystalline structure forming step (A-3) shown in section (c) of the present invention, the tape-attached test strip member 40 is immersed in a solution process containing a composition of a noble metal precursor and a reducing agent solution. At this point, a polycrystalline structure composed of multiple nanoparticle clusters and having multiple grain boundaries is formed on the test strip member 40 excluding the tape-attached portion. The portion formed with this polycrystalline structure becomes the detection portion of the lateral flow chromatography test strip.

[0149] Although not limited thereto, the polycrystalline structure is formed by a solution process using a composition including a noble metal precursor and a reducing agent solution with a ratio of the reducing agent to the noble metal precursor of 1:0.5 to 1:15.

[0150] Although not limited thereto, if the ratio of the reducing agent to the noble metal precursor is less than 1:0.5, a multi-wavelength Raman spectroscopy system cannot be utilized; and if it is greater than 1:15, it may be difficult to obtain a lateral flow chromatography test strip with excellent signal intensity and signal uniformity due to the film-forming effect.

[0151] Therefore, in the composition of the present invention, the ratio of the reducing agent to the noble metal precursor is 1:0.5 to 1:15, suitable for producing a substrate for spectral analysis with excellent signal intensity and signal uniformity. Specifically, the ratio may be 1:0.5 to 1:12, 1:0.5 to 1:10, 1:1 to 1:10, 1:1 to 1:9, 1:1 to 1:8, 1:1 to 1:7, 1:1 to 1:6, 1:1 to 1:5, 1:1 to 1:4, or 1:1 to 1:3. Furthermore, this range may vary slightly depending on the type of reducing agent.

[0152] Although not limited thereto, for the composition of the present invention, as the ratio of the reducing agent relative to the noble metal precursor increases, the size of the polycrystalline structure formed on the substrate for spectral analysis may decrease, and the density of the polycrystalline structure may increase.

[0153] Although not limited thereto, the nanoparticles may be composed of at least one of Au, Ag, and Pt, wherein Au is most suitable for improving signal intensity and signal uniformity.

[0154] Although not limited thereto, the noble metal precursor may be selected from the group consisting of HAuCl 4 , AuCl, AuCl 2 , AuCl 3 , Na 2 Au 2 Cl 8 , and NaAuCl 2 , wherein one or more of HAuCl 4 and NaAuCl 4 may be more suitable.

[0155] Although not limited thereto, the reducing agent may be one or more of hydroxylamine (HA), ascorbic acid (AA), FeSO 4 , and hydroxyquinone (HQ).

[0156] In such Figure 16 The tape removal step (A-4) shown in part (d) is a step of removing the tape 42 outside the polycrystalline structure formation area. Figure 16 As shown in part (e) of FIG. 4 , the first detection portion 22 and the second detection portion 24 including a polycrystalline structure are formed on the test strip member 40 .

[0157] Figure 17 Comparative images showing the shapes of polycrystalline structures formed by different types of reducing agents when CA is used as a test strip component when a lateral flow chromatography test strip including a polycrystalline structure is manufactured using a method for manufacturing a polycrystalline structure (method A) according to an embodiment of the present invention.

[0158] like Figure 17 As shown, under the same CA test strip structure, different reducing agents HA ( Figure 17 (a) of the HQ( Figure 17 Part (b) of) or AA( Figure 17 In the case of (c) of the reaction mixture, it can be confirmed that the shapes of the formed polycrystalline structures slightly differ depending on the type of reducing agent, but the polycrystalline structures are all well formed.

[0159] Figure 18 Comparative images showing the shapes of polycrystalline structures formed from different types of strip components when manufactured using the same reducing agent (HA) using a method for manufacturing a lateral flow chromatography test strip including a polycrystalline structure according to an embodiment of the present invention (Method A).

[0160] like Figure 18 As shown, under the same HA reducing agent, different test strip components CA paper ( Figure 18 Part (a) of MCE paper ( Figure 18 (b) of the NC paper ( Figure 18 (c) of the CN95 paper) or Figure 18 In the case of (d) of FIG. 1 ), it can be confirmed that the shapes of the formed polycrystalline structures slightly differ depending on the type of test strip member, but the polycrystalline structures are all well formed.

[0161] Figure 19 To schematically illustrate a method (Method B), a polycrystalline structure is formed according to one embodiment of the present invention and then a lateral flow chromatography test strip including the polycrystalline structure is manufactured in a patterning manner.

[0162] like Figure 19 As shown, according to another aspect of the present invention, a method for manufacturing a lateral flow chromatography test strip includes: a test strip component preparation step B-1) of preparing a test strip component 40 for lateral flow chromatography; a polycrystalline structure formation step B-2) of immersing the test strip component 40 in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure 44 composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip component; a tape attaching step B-3) of attaching a tape 42 to an area of the test strip component 40 other than an area to be patterned; and a tape removal step B-4) of removing the tape 42 from the test strip component 40.

[0163] Compared to Figure 16 The manufacturing method of the lateral flow chromatography test strip shown in FIG. 1 (Method A) is different in that the polycrystalline structure 44 is formed on the test strip member 40 and then patterned by attaching the tape 42. Figure 16 The same parts of the manufacturing method of the lateral flow chromatography test strip are omitted in detail.

[0164] like Figure 19 As shown in part (b) of FIG. 1 , in the polycrystalline structure forming step B-2) of forming the polycrystalline structure, the polycrystalline structure 44 is formed over the entire area of the test strip member 40 .

[0165] like Figure 19 As shown in part (c), in the tape attaching step B-3), patterning is achieved by attaching a tape 42 outside the area where the detection portion is to be formed.

[0166] like Figure 19 As shown in part (d) of FIG. 4 , in the tape removal step B-4), the polycrystalline structure in the adhesive area of the tape 42 is removed together with the tape due to the adhesive force of the tape 42. Figure 19 As shown in part (e) of FIG. 1 , a first detection portion 22 and a second detection portion 24 including two or more detection lines including a polycrystalline structure are formed on the test strip member.

[0167] Figure 20 Comparative images showing the shapes of polycrystalline structures formed by different types of reducing agents when CA is used as a test strip component in a lateral flow chromatography test strip comprising a polycrystalline structure manufactured by a method for manufacturing a polycrystalline structure (method B) according to one embodiment of the present invention.

[0168] like Figure 20 As shown, under the same CA test strip structure, different reducing agents HA ( Figure 20 (a) of the HQ( Figure 20 Part (b) of) and AA( Figure 20 In the case of (c) of the reaction mixture, it can be confirmed that the shapes of the formed polycrystalline structures slightly differ depending on the type of reducing agent, but the polycrystalline structures are all well formed.

[0169] Figure 21 The present invention illustrates a method for manufacturing a lateral flow chromatography test strip comprising a polycrystalline structure by forming the polycrystalline structure and then patterning the polycrystalline structure according to an embodiment of the present invention, and compares the heights of the detection portion of the test strip component formed according to different reducing agent types and reducing agent mixing ratios.

[0170] like Figure 21 As shown, it can be confirmed that the height of the detection portion varies depending on the type of reducing agent and the mixing ratio of the reducing agent.

[0171] Figure 22 A schematic diagram schematically illustrates a method (method C) for manufacturing a lateral flow chromatography test strip including a foreign material detection portion according to an embodiment of the present invention, wherein the foreign material detection portion has a detection line including a polycrystalline structure and a detection line including graphene oxide.

[0172] Reference Figure 22 According to another aspect of the present invention, a method for manufacturing a lateral flow chromatography test strip includes: a test strip component preparation step C-1) of preparing a test strip component 40 for lateral flow chromatography; a graphene oxide layer formation step C-2) of forming a graphene oxide layer 46 on the test strip component 40 using a graphene oxide solution; a first tape attaching step C-3) of attaching a tape 42 to an area of the test strip component 40 other than an area where graphene oxide patterning is to be performed; and a first tape removal step C-4) of removing a graphene oxide layer from the test strip component 40. The adhesive tape 42 is removed from the test strip member 40; a second adhesive tape attaching step C-5) is to attach the adhesive tape 42 to an area of the test strip member 40 other than the area where the polycrystalline structure patterning is to be performed; a polycrystalline structure forming step C-6) is to immerse the test strip member 40 in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member 40; and a second adhesive tape removing step C-7) is to remove the adhesive tape 42 from the test strip member 40.

[0173] like Figure 22 Part (a) and Figure 22As shown in part (b) of FIG. 2 , the graphene oxide layer 46 can be formed by filtering a graphene oxide solution onto the test strip member 40 under vacuum conditions. Although not limited thereto, when forming the graphene oxide layer 46, a graphene oxide solution having a concentration of 0.1 mg / mL to 10 mg / mL is used to quench the fluorescence signal.

[0174] like Figure 22 As shown in part (c) of FIG. 5 , the first tape attaching step C-3) is a step of patterning the tape 42 to form a detection portion including a graphene oxide layer 46 .

[0175] like Figure 22 As shown in part (d) of FIG. 4 , the tape removal step C-4) is a step of removing the graphene oxide outside the region of the detection portion including the graphene oxide layer 46 using the tape 42 .

[0176] The second tape attaching step C-5) is a step of patterning the tape 42 to form a detection portion including a polycrystalline structure.

[0177] like Figure 22 As shown in part (e), the polycrystalline structure forming step C-6) of forming the polycrystalline structure is a step of forming a detection portion including the polycrystalline structure in a portion where the tape 42 is not attached.

[0178] like Figure 22 Part (f) and Figure 22 As shown in part (g), in the second tape removal step C-7), the tape 42 is removed, thereby forming the detection portion (the first detection portion 22 and the second detection portion 24) including the detection line of the foreign material.

[0179] According to yet another aspect, a method for manufacturing a lateral flow chromatography system of the present invention (Method D) includes: a) a test strip member preparation step of preparing a test strip member for lateral flow chromatography; b) a polycrystalline structure formation step of immersing the test strip member in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member; c) a first tape attaching step of attaching a tape to an area of the test strip member other than an area where the polycrystalline structure is to be patterned; d) a first tape removal step of removing the tape from the test strip member; e) a second tape attaching step of attaching the tape to an area of the test strip member other than an area where graphene oxide is to be patterned; f) a graphene oxide layer formation step of forming a graphene oxide layer on the test strip member using a graphene oxide solution; and g) a second tape removal step of removing the tape from the test strip member.

[0180] The method D and Figure 22The difference of the method C shown is that the detection portion including the nanoparticle structure is formed before the detection portion including the graphene oxide layer.

[0181] Figure 23 Part (a) is based on Figure 22 Scanning electron microscopy (SEM) image of the polycrystalline nanostructured detection portion of the fabricated lateral flow test strip. Figure 23 Part (b) is based on Figure 22 Scanning electron microscopy (SEM) image of the graphene oxide detection portion of the fabricated lateral flow chromatography test strip.

[0182] Reference Figure 23 , it can be confirmed that the polycrystalline nanostructure detection part and the graphene oxide detection part are well formed.

[0183] As described above, the method for manufacturing a lateral flow chromatography test strip of the present invention can pattern a polycrystalline structure by solution processing, thereby easily manufacturing multiple test lines and test lines of different materials, and effectively adjusting the distance between test lines.

[0184] Although one embodiment of the present invention has been described above, those skilled in the art may make various modifications and changes to the present invention by adding, transforming, deleting or adding constituent elements without departing from the scope of the technical idea of the present invention as described in the claims, and this should also be deemed to be included in the scope of the rights of the present invention.

[0185] Description of Reference Numerals

[0186] 1: Lateral flow test strips

[0187] 10: Sample area

[0188] 20: Testing Area

[0189] 22, 24, 24a, 24b, 24c: Detection unit

[0190] 26: Hydrophobic treatment area

[0191] 30: Absorption area

[0192] 40: Test strip component

[0193] 42: Tape

[0194] 44: Polycrystalline structure

[0195] 46: Graphene oxide layer

Claims

1. A lateral flow chromatography test strip, wherein: The lateral flow chromatography test strip comprises a test strip member, wherein the test strip member comprises a test area, wherein the test area comprises one or more detection parts where a biochemical reaction occurs between a target contained in the sample and a probe when the sample flows by capillary action. Furthermore, the detection portion includes a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries.

2. The lateral flow test strip according to claim 1, wherein The test strip member is configured in one direction and includes: a sample area for introducing a sample; the test area; and Absorb excess sample to maintain the lateral flow absorption area.

3. The lateral flow test strip according to claim 1, wherein The lateral flow chromatography test strip is used for surface enhanced Raman spectroscopy, fluorescence or plasmon enhanced fluorescence analysis.

4. The lateral flow test strip according to claim 1, wherein The nanoparticles are composed of one or more of Au, Ag and Pt.

5. The lateral flow test strip according to claim 1, wherein The polycrystalline structure is combined with a gene probe.

6. The lateral flow test strip according to claim 1, wherein The detection unit includes one or more of the following: i) Two or more vertical detection lines perpendicular to the sample flow direction; ii) two or more horizontal detection lines parallel to the direction of sample flow; and iii) one or more vertical detection lines perpendicular to the sample flow direction and one or more horizontal detection lines horizontal to the sample flow direction.

7. The lateral flow test strip according to claim 1, wherein The detection portion includes one or more vertical detection lines perpendicular to the sample flow direction and one or more circular, elliptical or polygonal detection portions located behind the vertical detection lines relative to the sample flow direction.

8. The lateral flow test strip according to claim 7, wherein A hydrophobic treatment member is locally provided around the detection portion so that the sample passing through the vertical detection line is collected at the detection portion.

9. The lateral flow test strip according to claim 1, wherein The detection portion further includes a detection portion containing a foreign material.

10. The lateral flow test strip according to claim 9, wherein The detection unit includes: a first detection portion, forming a graphene oxide layer, and Two or more second detection parts forming the polycrystalline structure; The first detection part is combined with a gene probe labeled with a fluorescence quenching fluorescent molecule, The second detection part is combined with a probe for the fluorescent molecule to bind, and the fluorescent molecule restores fluorescence when the gene probe binds to the miRNA target molecule. The lateral flow test strip can thus be used as a multiplex sensor.

11. The lateral flow test strip according to claim 10, wherein The probe attached to each of the second detection parts is anti-FAM, anti-TAMRA, anti-methylene blue, anti-Cy5, anti-Cy3, anti-AlexaFluor 488, anti-Rhodamine Red, anti-Texas Red or anti-fluorescein / Oregon Green.

12. The lateral flow test strip according to claim 1, wherein The test strip component is composed of one or more of cellulose acetate, mixed cellulose esters, nitrocellulose, CN95 and a filter membrane formed by a synthetic polymer.

13. The lateral flow test strip according to claim 1, wherein The sample is selected from one or more of cells, metabolites, proteins, nucleic acids, DNA, RNA, enzymes, organic molecules, viruses, extracellular vesicles, microvesicles, exosomes and fat in urine, saliva, sweat, blood and tears.

14. The lateral flow test strip according to claim 1, wherein The detection portion comprises a first detection portion and a second detection portion arranged in sequence relative to the sample flow direction, The polycrystalline structure of the first detection portion that first contacts the sample is bound to a probe peptide that is dissociated by matrix metalloproteinase. A probe for detecting the dissociated probe peptide is bound to the second detection portion. The lateral flow test strip can thus be used as a matrix metalloproteinase sensor.

15. The lateral flow test strip according to claim 1, wherein The detection portion includes a first detection portion relative to the sample flow direction, and a second detection portion and a third detection portion located behind the first detection portion, wherein the second detection portion and the third detection portion are parallel to each other. The polycrystalline structure of the first detection portion is bound to a gene that reacts with an intercalating dye as a first control. The polycrystalline structure of the second detection part is bound to an antibody that can bind to Me-DNA. The polycrystalline structure of the third detection part is combined with the gene probe, The lateral flow test strip can thus be used as a Me-DNA sensor.

16. The lateral flow test strip according to claim 1, wherein The detection portion comprises a first detection portion, a second detection portion and a third detection portion which are arranged horizontally relative to the sample flow direction, and the first detection portion, the second detection portion and the third detection portion are parallel to each other. The polycrystalline structure of the first detection portion is bound to a gene that reacts with an intercalating dye as a first control. The polycrystalline structure of the second detection part is bound to an antibody that can bind to Me-DNA. The polycrystalline structure of the third detection part is combined with the gene probe, The lateral flow test strip can thus be used as a Me-DNA sensor.

17. A method for manufacturing a lateral flow chromatography test strip, in, include: Test strip component preparation step A-1): preparing a test strip component for lateral flow chromatography; Adhesive tape attaching step A-2), attaching the adhesive tape to an area of the test strip member other than the area to be patterned; a polycrystalline structure forming step A-3) of immersing the test strip member in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member; and Adhesive tape removal step A-4), removing the adhesive tape from the test strip member.

18. A method for manufacturing a lateral flow chromatography test strip, in, include: Test strip component preparation step B-1): preparing a test strip component for lateral flow chromatography; a polycrystalline structure forming step B-2) of immersing the test strip member in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member; Adhesive tape attaching step B-3), attaching the adhesive tape to the area of the test strip member other than the area to be patterned; and Adhesive tape removal step B-4), removing the adhesive tape from the test strip member.

19. A method for manufacturing a lateral flow chromatography test strip, in, include: Test strip component preparation step C-1): preparing a test strip component for lateral flow chromatography; a graphene oxide layer forming step C-2) of forming a graphene oxide layer on the test strip member using a graphene oxide solution; a first tape attaching step C-3) attaching the tape to an area of the test strip member other than the area where graphene oxide patterning is to be performed; a first tape removal step C-4), removing the tape from the test strip member; A second tape attaching step C-5) attaching the tape to an area of the test strip member other than the area where the polycrystalline structure is to be patterned; a polycrystalline structure forming step C-6) of immersing the test strip member in a composition containing a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member; and A second tape removal step C-7) removes the tape from the test strip member.

20. A method for manufacturing a lateral flow chromatography test strip, in, include: Test strip component preparation step D-1), preparing a test strip component for lateral flow chromatography; a polycrystalline structure forming step D-2) of immersing the test strip member in a composition comprising a noble metal precursor and a reducing agent solution to form a polycrystalline structure composed of a plurality of nanoparticle clusters and having a plurality of grain boundaries on the test strip member; A first tape attaching step D-3) attaching the tape to an area of the test strip member other than the area where the polycrystalline structure is to be patterned; a first tape removal step D-4), removing the tape from the test strip member; a second tape attaching step D-5) attaching the tape to an area of the test strip member other than the area where graphene oxide patterning is to be performed; a graphene oxide layer forming step D-6) of forming a graphene oxide layer on the test strip member using a graphene oxide solution; and A second tape removal step D-7) is to remove the tape from the test strip member.

Images