TW201643431A - Quantitative lateral flow chromatographic structure - Google Patents

Abstract

The invention relates to a quantitative lateral flow chromatographic structure. The quantitative lateral flow chromatographic structure has an elongated housing, and the housing is provided with a window having a reaction area and a correction area built therein along the longitudinal direction. A specimen collection area is extended from the outside of the window for collecting a specimen to be tested. The main technical features comprises the following steps: dropping the specimen to be tested into the quantitative lateral flow chromatographic structure through the specimen collection area; sequentially coating the specimen on a plurality of labeled reagents of the reaction area and the correction area for reaction by a capillary phenomenon; and forming a complex of a plurality of specimens to be tested to proceed with an immunochromatographic test. The labeled reagents include a first labeled reagent sprayed and coated on the reaction area and a second labeled reagent sprayed and coated on the correction area. The first labeled reagent has an antibody which is totally different form an antibody of the second labeled reagent.

Description

Lateral flow chromatography quantitative structural mechanism

The invention relates to a quantitative flow mechanism of lateral flow chromatography, in particular to an immune reaction detection mechanism for simultaneously detecting two different antibodies for detecting a sample, mainly by using a window of a lateral flow chromatography quantitative device. Spraying two different types of antibodies on the reaction zone and the calibration zone, and simultaneously absorbing the same sample for immunoreactivity detection, can effectively prevent the situation that the traditional antibody only interferes with each other, thereby improving the quantitative detection of immunochromatography technology. Accuracy.

According to the immunochromatographic quantitative detection technology, a detection method in which an antigen and an antibody react with each other is used, that is, a prepared specific antigen or antibody is used as an analyte to detect a corresponding antibody or antigen in the sample; immunochromatography The main feature is high specificity and sensitivity. If the markers for analyzing antigens or antibodies (such as radionuclides, luciferins, enzymes, latex, immunogold, carbon black, etc.) are labeled, After the corresponding antibody or antigen in the body is reacted, the antigen-antibody complex itself can be determined without determining the label in the complex, and the sensitivity of the immunological technique can be greatly improved by the amplification of the label; immunochromatographic detection (immunochromatographic) Test (referred to as ICT) has played a very important role in today's medical system. It can be used to qualitatively or quantitatively measure changes in various biological indicators in human body fluids to provide medical personnel with information for disease diagnosis or drug treatment. Chromatographic detection (ICT) detects immune responses in a simple, single-step manner in a very short time The tester can directly interpret the test results with the naked eye, mainly by causing the solution to flow in the water-absorbent fiber layer with different tightness by capillary phenomenon, and using the immunoaffinity between the test substance and the analyte to interpret The markers on the analyte are used to qualitatively, semi-quantitatively or quantitatively detect changes in various biological indicators for detection purposes.

Since the immunochromatographic assay utilizes the specificity of the antigen-antibody reaction to identify and detect antigens or antibodies; conventionally used immunochromatographic detection structures respectively spray a reaction line on a fiber membrane in a window. A quality control C line (control line), and the reaction T line and the quality control C line are sprayed with the same antibody, so that when the immunochromatographic detection of the sample is performed, the sample to be tested has been reacted first. When the line is absorbed, the sample to be flowed to the quality control C line with the same antibody as the reaction T line, often has a good reaction with the quality control C line due to insufficient reaction amount, and affects the correctness of the result interpretation; therefore, How to effectively overcome the problem of mutual interference between the reaction T line and the quality control C line, and then strengthen the accuracy of the quantitative detection of immunochromatography technology, is still a problem that researchers of medical equipment and other related academic units need to continuously overcome and solve.

Nowadays, the inventor is a tireless spirit in view of the fact that the reaction zone and the calibration zone of the above-mentioned conventional immunochromatography technology interfere with each other due to the use of the same antibody, and by virtue of its rich professional knowledge and many years of The practical experience assisted and improved, and the present invention was developed based on this.

The main object of the present invention is to provide a side-stream chromatography quantitative structural mechanism, in particular to an immunoreactive detection mechanism for simultaneously detecting two different antibodies for detecting a sample, mainly by a window of a lateral flow chromatography quantitative device. Spraying two different types of antibodies on the reaction zone and the calibration zone, and simultaneously absorbing the same sample for immunoreactivity detection, can effectively prevent the situation that the traditional antibody only interferes with each other, and achieves quantitative detection by immunochromatography Accuracy.

In order to achieve the above-mentioned implementation object, the present inventors propose a side-flow chromatography quantitative structural mechanism, which is mainly realized by a one-side flow chromatography quantitative device, and the lateral flow chromatography quantitative device has a long strip-shaped outer casing, and the outer casing is along The length of the window extends with a window for constructing a reaction zone and a correction zone, and the exterior of the window extends with a sample collection area for collecting the sample to be detected. The main technical feature is that the sample to be tested passes the test. The body collection area is dropped into the lateral flow chromatography quantitative device, and is sequentially reacted with a plurality of labeled reagents coated on the reaction zone and the correction zone by capillary flow to form a plurality of analyte complexes. Immunochromatographic quantitative detection, and the labeled reagent comprises a first labeled reagent sprayed on the reaction zone, and a second labeled reagent sprayed on the calibration zone, wherein the first labeled reagent and the first The two-band label reagent contains two antibodies that are completely different.

The lateral flow chromatography as described above quantifies the structural mechanism, wherein the label carried by the first labeling reagent is one of colloidal gold or latex particles and the like.

The lateral flow chromatography as described above quantifies the structural mechanism, wherein the species source of the first labeled reagent comprises one of mouse, sheep, goat, rabbit, duck or scorpion antibody.

The lateral flow chromatography as described above quantifies the structural mechanism, wherein the label carried by the second labeled reagent is one of colloidal gold or latex particles and the like.

The lateral flow chromatography as described above quantifies the structural mechanism, wherein the source of the second labeled reagent includes a mouse, a sheep, a goat, a rabbit, a duck or a donkey antibody, but is different from the first labeled reagent. The source of the species used.

The lateral flow chromatography as described above quantifies the structural mechanism, wherein the sample collection zone, the reaction zone, and the calibration zone are composed of a water-absorbing fibrous layer.

The lateral flow chromatography quantitative structure mechanism as described above, wherein the lateral flow chromatography quantitative device can be further combined with a detection machine for immunochromatographic detection, the accuracy of the detection is due to the first label reagent and the second label The reagents are different and do not interfere with each other.

Thereby, the side-stream chromatography quantitative structural mechanism of the present invention absorbs the same by simultaneously spraying two completely different types of antibody-labeled reagents on the reaction zone and the calibration zone in the window of the lateral flow chromatography quantitative device. The detection of the immunological reaction of the sample to be detected, the accuracy of the detection may be different from the labeling reagents of two completely different types of antibodies without interfering with each other, thereby effectively achieving the advantage of improving the accuracy of quantitative detection by immunochromatography In addition, the sidestream chromatography quantitative structural mechanism of the present invention is different from the source of the species of the second labeled reagent sprayed on the correction zone by the first labeled reagent sprayed on the reaction zone, respectively An independent design of one of the mouse, sheep, goat, rabbit, duck or scorpion antibody, such that the second labeled reagent on the calibration zone reacts with the sample to be tested to eliminate the first species of different species in the reaction zone. The interference with the labeling reagent can effectively prevent the situation that the traditional antibody only interferes with each other, and the detection accuracy of the immunochromatographic technique is improved. of.

The object of the present invention and its structural design and advantages will be explained in the light of the preferred embodiments shown in the following drawings, so that the reviewing committee can have a more in-depth and specific understanding of the present invention.

First of all, in order to better understand the technical features of the present invention, the basic concept of "immunocolloidal gold technology" will be briefly explained. The immunocolloidal gold technology is an immunological method which is widely used in various fields of biomedicine and is available. The detection of antibodies such as HBsAg, hCG or anti-double-stranded DNA is simple, rapid, accurate, and non-polluting; chloroauric acid can be polymerized into a certain size of gold particles under the action of a reducing agent to form a negatively charged hydrophobic Glue solution, which is a stable colloidal state due to electrostatic action, is called colloidal gold; it uses a negatively charged property of colloidal gold in an alkaline environment, and forms a strong bond with the positive charge group of the protein molecule by electrostatic attraction. In addition to antibody proteins, colloidal gold can also bind to a variety of other biomacromolecules, depending on some physical properties of the colloidal gold, such as high electron density, particle size, shape, and color response, plus the immunobiological properties of the conjugate. It has been widely used in immunology, histology, pathology and cell biology marker research.

Immunogold labeling techniques are commonly used to detect infectious diseases (such as sexually transmitted diseases or hepatitis), tumors (such as colorectal cancer or nasopharyngeal cancer), endocrine (such as pregnancy or ovulation), drug abuse (ecstasy) or blood type identification, many The rapid test reagent is manufactured by immunological techniques. In the case of pregnancy test, a pregnant woman's body produces a hormone that is much higher than usual. If a pregnant woman's urine is sampled in a reagent, in the urine. A high amount of HCG hormone binds to the anti-HCG antibody labeled with colloidal gold, binds to the corresponding position on the substrate, and exhibits a red to purple color depending on the size of the colloidal gold. Such a color exhibits an optical effect from colloidal gold. When part of the colloidal gold size is much smaller than the wavelength of light, the visible light scattering spectrum will have a distinct resonance phenomenon, called surface plasma resonance. The so-called plasma (plasmon) means that the free electron cloud in the colloidal gold is going to the same moment. Moving in the same direction, if the orderly motion frequency is the same as the specific applied light frequency, resonance will occur, and the applied light energy will be effectively transferred to Ordered motion of the electron cloud, and therefore, this frequency light waves will be efficiently absorbed, to form a unique color.

Next, referring to the first figure, an external perspective view of a preferred embodiment of the lateral flow chromatography quantitative structure mechanism of the present invention, wherein the lateral flow chromatography quantitative structural mechanism of the present invention is mainly performed by one-side flow chromatography The quantitative device (1) realizes that the lateral flow chromatography quantitative device (1) has a long outer casing (11), and the outer casing (11) extends along the length thereof to have a built-in reaction zone (121) and a The window (12) of the correction area (122), and the external part of the window (12) is extended with a sample collection area (13) for collecting the sample (2) to be detected, and the main technical feature is that the test is to be performed. The body (2) is dropped into the lateral flow chromatography quantitative device (1) via the sample collection area (13), and sequentially applied to the reaction zone (121) and the correction zone (122) by capillary flow. A plurality of labeled reagents are reacted to form a plurality of analyte complexes for quantitative detection by immunochromatography, and the labeled reagents comprise a first labeled reagent (1211) sprayed on the reaction zone (121). And a second labeled reagent (1221) sprayed onto the calibration zone (122), wherein the first labeled reagent (1211) is Two labeled reagents (1221) system comprises two completely different antibodies.

In addition, the label of the first labeled reagent (1211) is one of colloidal gold or latex particles, and the source of the first labeled reagent (1211) may include mice, sheep, and goats. And one of the rabbit, duck or scorpion antibody; further, the label of the second labeling reagent (1221) may also be one of colloidal gold or latex particles, and the second labeling reagent ( The source of the species of 1221) may also include one of mouse, sheep, goat, rabbit, duck or scorpion antibody, but different from the species source used by the first labeling reagent (1211); using colloidal gold as an indicator Luteinizing hormone (LH) rapid test reagent is an example. It is developed by the principle of immunochromatography. Its function can be used for the detection of female ovulation. When the specimen (2) contains the progesterone antigen, it will be related to the main The colloidal gold lutein antibody on the reaction zone (111) binds to form an antigen-antibody-colloidal gold complex, and the sample containing the complex (2) flows to the window (12) via capillary phenomenon. a region (121), and an antibody-antigen-antibody sandwich is formed with another luteinizing hormone antibody immobilized at the position of the reaction region (121), and the reaction region (121) exhibits a purple-red reaction T-line with a correction region at the same time. (122) Another quality control C line is used as a control. To ensure the validity of the test reagent, any valid sample (2) will cause a red quality control C line to appear in the lateral flow chromatography device (1). The formation of the T-line of the quality control C line of the calibration zone (122) and the reaction zone (121) can also be visually observed to determine the effectiveness of the immune response and the test results.

In addition, the lateral flow chromatography quantitative device (1) of the present invention can be further combined with a detection machine (not shown) for immunochromatographic detection, and the accuracy of the detection is due to the first labeled reagent (1211) and the first The two-label reagents (1221) are different and do not interfere with each other; further, the sample collection area (13), the reaction area (121), and the correction area (122) are composed of a water-absorbing fibrous layer, The sample (2) such as human blood, urine, saliva or body fluid extract collected during the detection can be easily attached thereto by dropping the sample collection area (13) into the lateral flow chromatography quantitative device (1). And the water-absorbing fibrous layer is diffused to the reaction zone (121) and the calibration zone (122) for the subsequent immunological reaction test.

According to the above-mentioned lateral flow chromatography quantitative structure, in actual implementation, please refer to the second figure, which is a schematic diagram of the use of a preferred embodiment of the lateral flow chromatography quantitative structure mechanism of the present invention; When the examiner performs the immune response test, the blood, urine, saliva or body fluid extract of the subject must first be collected as a sample (2); then, the collected sample (2) is dropped into the lateral flow chromatography. The sample collection area (13) of the device (1), wherein the lateral flow chromatography quantitative structure mechanism of the present invention is mainly realized by a one-side flow chromatography quantitative device (1), and a lateral flow chromatography quantitative device (1) The utility model has a long outer casing (11), and the outer casing (11) extends along a length thereof to extend a window (12) for constructing a reaction zone (121) and a correction zone (122), and the window (12) The external system extends with a sample collection area (13) for collecting the sample (2) to be detected; and, because the material of the sample collection area (13) is a water-absorbing fibrous layer, the immune reaction can be detected. The sample (2) is easily attached thereto by dropping in the sample collection area (13), and is provided by a water-absorbent fiber layer. The fine phenomenon is diffused into the main reaction zone (111) to facilitate the subsequent immunological reaction; afterwards, the reaction zone (121) and the calibration zone (122) in the window (12) are also composed of a water-absorbing fibrous layer. The body (2) is also reacted by the capillary reaction phenomenon by the main reaction zone (111), and sequentially reacts with a plurality of labeled reagents coated on the reaction zone (121) and the correction zone (122) to form a plurality of The sample complex is subjected to immunochromatographic quantitative detection by a detection machine, wherein the labeled reagent comprises a first labeled reagent (1211) sprayed on the reaction zone (121), and sprayed on the calibration zone (122). The second labeled reagent (1221), and the first labeled reagent (1211) and the second labeled reagent (1221) contain two antibodies that are completely different, that is, the sample (2) First reacted with the first labeled reagent (1211) located on the reaction zone (121), and then reacted with the second labeled reagent (1221) located on the calibration zone (122), due to the first labeled reagent (1211) and the second labeled reagent (1221) belong to two completely different antibodies, so there is no mutual interference. Can effectively enhance the accuracy of quantitative detection of immune chromatography.

It can be seen from the above description that the side stream chromatography quantitative structure mechanism of the present invention has the following advantages compared with the prior art:

1. The sidestream chromatography quantitative structural mechanism of the present invention is to simultaneously absorb the same type by spraying two kinds of labeled reagents of completely different types of antibodies on the reaction zone and the calibration zone in the window of the lateral flow chromatography quantitative device. The detection of the test body for the detection of the immune response, the accuracy of the detection may be different due to the different labeling reagents of two completely different types of antibodies without interfering with each other, effectively achieving the advantage of improving the accuracy of quantitative detection of immunochromatographic techniques.

2. The sidestream chromatography quantitative structural mechanism of the present invention is different from the source of the species of the second labeled reagent sprayed on the correction zone by the first labeled reagent sprayed on the reaction zone, respectively. An independent design of one of the sheep, goat, rabbit, duck or scorpion antibody, such that the second label reagent on the calibration zone reacts with the sample to be detected to eliminate the first band of different species in the reaction zone The interference of the marker reagent can effectively prevent the situation that the traditional antibody only interferes with each other, and the purpose of improving the detection accuracy of the immunochromatography technology is achieved.

In summary, the side stream chromatography quantitative structure mechanism of the present invention can achieve the intended use efficiency by the above disclosed embodiments, and the present invention has not been disclosed before the application, and has completely complied with the patent law. Regulations and requirements.爰Issuing an application for a patent for invention in accordance with the law, and asking for a review, and granting a patent, is truly sensible.

The illustrations and descriptions of the present invention are merely preferred embodiments of the present invention, and are not intended to limit the scope of the present invention; those skilled in the art, which are characterized by the scope of the present invention, Equivalent variations or modifications are considered to be within the scope of the design of the invention.

(1) ‧‧‧ Lateral flow chromatography

(11) ‧‧‧ Shell

(111)‧‧‧Main reaction zone

(12)‧‧‧Window

(121) ‧‧‧Reaction zone

(1211)‧‧‧First Labeling Reagents

(122)‧‧‧ calibration area

(1221)‧‧‧Second label reagent

(13) ‧‧‧sample collection area

(2) ‧ ‧ ‧ specimens

First Figure: Appearance perspective view of a preferred embodiment of the side stream chromatography quantitative structure mechanism of the present invention

Second Figure: Schematic diagram of the use of a preferred embodiment of the side stream chromatography quantitative structure mechanism of the present invention

(1) ‧‧‧ Lateral flow chromatography

(11) ‧‧‧ Shell

(12)‧‧‧Window

(121) ‧‧‧Reaction zone

(1211)‧‧‧First Labeling Reagents

(122)‧‧‧ calibration area

(1221)‧‧‧Second label reagent

(13) ‧‧‧sample collection area

Claims (7)

A lateral flow chromatography quantitative structural mechanism is mainly realized by a one-side flow chromatography quantitative device having a long strip-shaped outer casing, the outer casing extending along its length has a built-in one a reaction window and a correction area window, and the external portion of the window extends with a sample collection area for collecting the sample to be detected, and the main technical feature is that the sample to be tested is dropped into the sample collection area through the sample collection area. In the lateral flow chromatography quantitative device, a plurality of labeled reagents coated on the reaction zone and the calibration zone are sequentially reacted by capillary flow to form a plurality of analyte complexes for immunochromatography. Quantitative detection, wherein the labeled reagent comprises a first labeled reagent sprayed onto the reaction zone, and a second labeled reagent sprayed onto the calibration zone, wherein the first labeled reagent The second labeled reagent reagent contains two antibodies that are completely different from each other. The lateral flow chromatography quantitative structural mechanism according to claim 1, wherein the label of the first labeled reagent is one of colloidal gold or latex particles. The lateral flow chromatography quantitative structural mechanism of claim 2, wherein the first source of the marker reagent comprises a mouse, sheep, goat, rabbit, duck or scorpion antibody. The lateral flow chromatography quantitative structural mechanism according to claim 1, wherein the label of the second labeled reagent is one of colloidal gold or latex particles. The lateral flow chromatography quantitative structural mechanism according to claim 4, wherein the second marker reagent source is one of a mouse, a sheep, a goat, a rabbit, a duck or a donkey antibody, but different from The source of the species used for the first labeled reagent. The lateral flow chromatography quantitative structure mechanism according to claim 1, wherein the sample collection area, the reaction area, and the correction area are composed of a water-absorbing fibrous layer. The lateral flow chromatography quantitative structure mechanism according to claim 1, wherein the lateral flow chromatography quantitative device is further combined with a detection machine for immunochromatographic detection, and the accuracy of the detection is due to the first label The reagents are different from the second labeled reagent without interfering with each other.