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KR-20260063762-A - BLADDER CANCER DIAGNOSTIC KIT USING NMP22 AS A BIOMARKER

KR20260063762AKR 20260063762 AKR20260063762 AKR 20260063762AKR-20260063762-A

Abstract

The present invention relates to a bladder cancer diagnostic kit using NMP22 as a biomarker, comprising: a support corresponding to a base body of a diagnostic strip installed in the internal space of the diagnostic kit body; a sample pad attached to the upper part of the support, into which a sample corresponding to the urine of a diagnostic subject and a buffer are injected; a detection film attached to the upper part of the support in a manner sequentially connected to the sample pad; a detection (Test) line formed by coating on the detection film and having an anti-NMP22 (Nuclear Matrix Protein 22) antibody immobilized thereon; and a control (Control) line formed by coating on the detection film at a position spaced apart from the detection line and having a Chicken IgY antibody immobilized thereon; wherein the buffer comprises a first complex in which a Europium (Eu)-based fluorescent bead is combined with an anti-Chicken IgY antibody and a second complex in which a Europium (Eu)-based fluorescent bead is combined with an anti-NMP22 (Nuclear Matrix Protein 22) antibody.

Inventors

  • 오택규

Assignees

  • 주식회사 올메이드

Dates

Publication Date
20260507
Application Date
20241031

Claims (6)

  1. A support corresponding to the base body of a diagnostic strip that is accommodated and installed in the internal space of the diagnostic kit body; A sample pad attached to the upper part of the above support, into which a specimen corresponding to the urine of the diagnostic subject and a buffer are injected; A detection film attached to the upper part of the support in a manner sequentially connected to the sample pads above; A detection (test) line formed by coating on the above detection film and having an anti-NMP22 (Nuclear Matrix Protein 22) antibody immobilized thereon; and A control line formed by coating at a position spaced apart from the detection line of the detection membrane, on which a Chicken IgY antibody is immobilized; comprising The above buffer is characterized by comprising a first complex in which a europium (Eu)-based fluorescent bead is conjugated with an anti-Chicken IgY antibody, and a second complex in which a europium (Eu)-based fluorescent bead is conjugated with an anti-NMP22 (Nuclear Matrix Protein 22) antibody. Bladder cancer diagnostic kit using NMP22 as a biomarker.
  2. In paragraph 1, The detection film has the control line and the detection line formed sequentially based on the sample pad side, and The anti-NMP22 antibody included in the above detection line is prepared at a concentration of 1.8 mg/mL to 2.2 mg/mL, and The Chicken IgY antibody included in the above control line is characterized by being prepared at a concentration of 0.8 mg/mL to 1.2 mg/mL. Bladder cancer diagnostic kit using NMP22 as a biomarker.
  3. In paragraph 2, The europium-based fluorescent beads combined within the first and second complexes included in the buffer are characterized by having an excitation wavelength of 330 nm to 340 nm and an emission wavelength of 610 nm to 620 nm. Bladder cancer diagnostic kit using NMP22 as a biomarker.
  4. In paragraph 3, The first complex included in the buffer is characterized by being prepared by treating europium-based fluorescent beads with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide to prepare them to a state capable of antibody binding, and then reacting 1.3 to 1.7 parts by weight of an anti-Chicken IgY antibody with 10 parts by weight of the europium-based fluorescent beads prepared to a state capable of antibody binding through stirring. Bladder cancer diagnostic kit using NMP22 as a biomarker.
  5. In paragraph 3, The second complex included in the buffer is characterized by being prepared by treating europium-based fluorescent beads with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide to prepare them to a state capable of antibody binding, and then reacting 0.8 to 1.2 parts by weight of an anti-NMP22 antibody with 10 parts by weight of the europium-based fluorescent beads prepared to a state capable of antibody binding through stirring. Bladder cancer diagnostic kit using NMP22 as a biomarker.
  6. In paragraph 3, The first complex included in the buffer is provided in an amount of 0.8 to 1.2 parts by volume relative to the total volume of the sample and buffer injected into the injection pad, which is 100 parts by volume, and The second complex included in the buffer is characterized by being provided in an amount of 0.8 to 1.2 parts by volume relative to the total volume of 100 parts by volume of the sample and buffer injected into the injection pad. Bladder cancer diagnostic kit using NMP22 as a biomarker.

Description

Bladder cancer diagnostic kit using NMP22 as a biomarker The present invention relates to a bladder cancer diagnostic kit using NMP22 as a biomarker. Bladder cancer diagnostic technologies are broadly classified into imaging tests, cytological examinations, and biomarker detection methods. Each method is used to accurately diagnose bladder cancer, and recently, these diagnostic technologies are evolving in the form of diagnostic kits that can be conveniently used outside of hospitals. Since bladder cancer has a high treatment efficacy when detected early, there is a growing need for non-invasive yet highly accurate diagnostic technologies. Accordingly, detection technologies utilizing various biomarkers are playing an important role, and in particular, biomarkers such as NMP22 (Nuclear Matrix Protein 22) are establishing themselves as important indicators in the diagnosis of bladder cancer. Among these, technologies related to bladder cancer diagnosis using biomarkers are developing into a useful method for early diagnosis and patient monitoring because they can be performed non-invasively and conveniently by checking for the presence of specific biomarkers in urine to determine the presence of cancer. Representative examples include NMP22, BTA, and FISH tests. In particular, NMP22 is a nuclear matrix protein that is detected at high concentrations in the urine of bladder cancer patients and is utilized as a major biomarker for bladder cancer. Bladder cancer diagnostic technology in the form of diagnostic kits, which is being developed based on such biomarker detection methods, miniaturizes and automates existing laboratory-based detection methods to enable rapid detection of bladder cancer biomarkers in urine, thereby making diagnosis possible without visiting a hospital. Recently developed diagnostic kits possess high sensitivity and specificity, allowing for more efficient bladder cancer diagnosis. In this regard, prior art for providing a bladder cancer diagnostic kit configured to quantitatively determine pathological changes in bladder cancer by providing multiple detection lines for NMP22, UBC, and BTA biomarkers, and capable of increasing sensitivity and specificity by using quantum dots of different sizes for each biomarker, includes Chinese Registered Patent Publication CN 108593922, "Joint detection kit for measuring bladder cancer and preparation method thereof" (hereinafter referred to as "prior art"). However, in the case of existing bladder cancer diagnostic kit technologies utilizing NMP22 as a biomarker, including conventional technologies, interference between other fluorescent substances may occur during fluorescence detection, making accurate signal detection difficult. Furthermore, the detection sensitivity required for the actual diagnosis of bladder cancer is still low, resulting in problems of reduced diagnostic accuracy and reliability. FIG. 1 is a cross-sectional view illustrating the structure of a diagnostic strip of a bladder cancer diagnostic kit using NMP22 of the present invention as a biomarker. FIG. 2 is a diagram illustrating the process of forming a complex by binding an antibody to a europium-based fluorescent bead included in a buffer injected into a sample pad in a bladder cancer diagnostic kit using NMP22 of the present invention as a biomarker. Figure 3 is a graph showing the results of analyzing the change in fluorescence intensity according to NMP22 concentration using the bladder cancer diagnostic kit of the present invention that utilizes NMP22 as a biomarker. Figure 4 is a graph showing the results of quantifying fluorescence intensity according to NMP22 concentration using the bladder cancer diagnostic kit of the present invention that utilizes NMP22 as a biomarker. Preferred embodiments of the present invention will be described in more detail with reference to the attached drawings, provided that technical details that are already well known are omitted or compressed for the sake of brevity. 1. Description of a bladder cancer diagnostic kit using NMP22 as a biomarker Referring to FIGS. 1 and 2, the bladder cancer diagnostic kit (100) using NMP22 as a biomarker according to the present invention is formed such that a diagnostic strip, as shown in FIG. 1, is received and installed within a separate receiving space provided inside the entire housing of the diagnostic kit (100), which corresponds to the body of the diagnostic kit. More specifically, the diagnostic strip received and installed in the body of the diagnostic kit includes a support (110), a sample pad (120), a detection film (130), an absorption pad (140), a control line (150), and a detection line (160). Here, a sample injection port is formed on one side of the upper part of the diagnostic kit body to provide an open space into which a sample and buffer can be injected into the sample pad (120) of the diagnostic strip. In addition, a transparent diagnostic result observation window may be formed on