Composition for diagnosing tuberculosis and method for diagnosing tuberculosis on basis of change in optical characteristics

A diagnostic method and composition technology, which can be used in scientific instruments, biological tests, material testing products, etc., can solve the problems of not yet developed technology for diagnosing tuberculosis by fluorescent nano-substance-antibody complexes, and achieve the effect of low detection limit

Pending Publication Date: 2020-07-28
DXGEN CORP
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] However, a technique for diagnosing tuberculosis based on changes in the optical properties of fluorescent nanosubstance-antibody complexes has not yet been developed.

Method used

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  • Composition for diagnosing tuberculosis and method for diagnosing tuberculosis on basis of change in optical characteristics
  • Composition for diagnosing tuberculosis and method for diagnosing tuberculosis on basis of change in optical characteristics
  • Composition for diagnosing tuberculosis and method for diagnosing tuberculosis on basis of change in optical characteristics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1. Preparation of Mycobacterium tuberculosis-specific antigen CFP10 and its recombinant antibodies G2 and G3

[0052] Referring to Korean Registered Patent No. 1631054, the tuberculosis antigen C FP10 to be used in the examples and the two Group2 (G2, Group 2) and Group3 (G3, Group 3) antibody genes that can bind to it were expressed in Escherichia coli And carry out separation and purification for future use.

[0053] To this end, the control vector pET22, the CFP10 antigen expression vector pET22-CFP10, the G2 antibody expression vector pET22-GBP-CFP10G2, and the G3 antibody expression vector pET22-CFP10G3 were transformed into Escherichia coli BL21 (DE3) strain. Each strain was cultured at 37°C in 100ml LB (1% (w / v) tryptone, 1% NaCl, 0.5% Yeast extract (Yeast extract)) + ampicillin medium To OD 0.4, 0.1 mM IPTG was added for protein expression, and after culturing for 6 hours, strains were collected by centrifugation (3,500 rpm, 4° C., 10 minutes).

[0054...

Embodiment 2

[0056] Example 2. Preparation of "Metal Nanoparticle-Antibody Complex" and "Fluorescent Nanomaterial-Antibody Complex" against Mycobacterium tuberculosis-specific antigen CFP10

[0057] 2-1: Preparation of metal nanoparticle-antibody complex

[0058] 2-1-1: Preparation of Magnetic Nanoparticle-Antibody Complex

[0059] 2-1-1-1: Amine Functional Group Substitution of Magnetic Nanoparticles

[0060] After adding 9 mL of methanol to 1 mL of APTES solution and mixing well, 10 mg of magnetic nanoparticles were added to disperse with an ultrasonic mill, and stirred for 17 hours to form amine functional groups on the surface. Thereafter, the supernatant was discarded after the magnetic nanoparticles were collected by a magnet, washed three times with methanol (10 mL for each wash) and the magnet to prepare magnetic nanoparticles substituted by amine groups, and then stored at 4°C.

[0061] 2-1-1-2: Preparation of Magnetic Nanoparticle-G2 Antibody Complex

[0062]The magnetic nanop...

Embodiment 3

[0083] Example 3. Detection of Mycobacterium tuberculosis using "metal nanoparticle-antibody complex" and "fluorescent nanomaterial-antibody complex"

[0084] 3-1: Tuberculosis (TB) Detection Using Magnetic Nanoparticle-Antibody Complex and Upconverting Nanoparticle-Antibody Complex

[0085] Place 0.5 mL of the upswitching nanoparticle-G3 antibody complex prepared in 2-2-2 and 0.3 mL of the magnetic nanoparticle-G2 antibody complex prepared in 2-1-1-2 into a 1.5 mL microtube Mix thoroughly afterwards. Then, 0.05 mL of a solution containing tuberculosis antigen (CFP-10) was added, and after the reaction was carried out with gentle shaking at room temperature for 30 minutes, the upconverting nanoparticles bound by the tuberculosis antigen-magnetic nanoparticles and untreated nanoparticles were collected by a magnet. After the bound magnetic nanoparticles were discarded, the supernatant was discarded, and after washing 3 times with a 10 mM PBS (pH 7.4) solution (3 mL for each wa...

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Abstract

The present invention relates to a composition for diagnosing tuberculosis and a method for diagnosing tuberculosis on the basis of a change in optical characteristics. The method for diagnosing tuberculosis is characterized in that the method is based on a change in optical characteristics of (a) a fluorescent nanomaterial-antibody complex and a fluorescent nanomaterial-antibody complex or (b) afluorescent nanomaterial-antibody complex and a metal nanoparticle-antibody complex. The method for diagnosing tuberculosis on the basis of a change in optical characteristics according to the presentinvention has a low detection limit and can rapidly and accurately measure tuberculosis since a change in fluorescence intensity of metal nanoparticles and fluorescent nanomaterials is utilized.

Description

technical field [0001] The present invention relates to a composition for diagnosing tuberculosis and a method for diagnosing tuberculosis based on changes in optical properties. Background technique [0002] In general, tuberculosis is a disease with the highest incidence among notifiable infectious diseases in Korea, and is a disease caused by infection with Mycobacterium tuberculosis. Therefore, early diagnosis and treatment that can obtain reliable information from patients suspected of tuberculosis infection in a short period of time are more important than anything else. [0003] The tuberculin skin test (TST) method is a routine method for diagnosing tuberculosis, but because of its low specificity, it has low reliability in diagnosing tuberculosis. However, it is still widely used due to its advantages of high sensitivity and low cost. The culture method is also widely used. As the most reliable detection method among various tuberculosis diagnostic methods, and as...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/58G01N33/543
CPCG01N33/5695G01N33/587G01N33/54346G01N33/54326
Inventor 李进雨田善娥
Owner DXGEN CORP
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