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Tuberculosis immunodiagnosis molecular marker and application thereof in preparation of vaccines

A technology for tuberculosis and Mycobacterium tuberculosis, applied in the fields of immunology and molecular biology, can solve the problems of easy false positive rate, can not be used as a diagnostic basis, and easy to be misdiagnosed.

Active Publication Date: 2017-05-03
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is easy to be confused with other lung diseases and needs to be confirmed by a professional doctor
(3) Immunological diagnosis of pulmonary tuberculosis: 1. The tuberculin pure protein derivative (PPD) test is commonly used. A positive test is one of the evidences of tuberculosis infection, but the false positive rate is high and it is easy to misdiagnose
2. Positive tuberculosis antibody tests in blood and sputum are also helpful for diagnosis, and false positive rates are also prone to occur
4. Polymerase chain reaction (PCR), the advantage is that the sensitivity can reach 98%-100%, and the disadvantage is that the specificity is poor
(4) Other examinations: can only be used as an auxiliary diagnosis, not as a basis for diagnosis

Method used

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  • Tuberculosis immunodiagnosis molecular marker and application thereof in preparation of vaccines
  • Tuberculosis immunodiagnosis molecular marker and application thereof in preparation of vaccines
  • Tuberculosis immunodiagnosis molecular marker and application thereof in preparation of vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: Mass spectrometry identification experiment

[0047] 1. Experimental Materials and Instruments

[0048]Target protein samples: prepared Rv0237 protein (SEQ ID NO:1) and Rv1111c protein (SEQ ID NO:2).

[0049] Instruments: Ultimate 3000Nano-LC system (Dionex, USA), linear ion trap multistage tandem mass spectrometer (linear trap quadruple, LTQ), Orbitrap Velos mass spectrometer (ThermoScientific, Germany).

[0050] Data processing software: MassLynx version 4.1 (Dionex, USA), Xcalibur software v2.2.6 (Thermo), MASCOT version 2.2 (Matrix Sciences, UK).

[0051] 2. Experimental method

[0052] (1) After the target protein is subjected to SDS PAGE, rinse the film with deionized water for 15 minutes; cut off the target fragment, 1-2 mm in size; put it in a centrifuge tube with low protein adsorption, and use 100 μL of ddH 2 O Rinse the pellets in the dish and repeat, discard the liquid and add 40 μL of (ACN) / ddH 2 O(50 / 50), discard the liquid, incubate for...

Embodiment 2

[0075] Embodiment 2: Humoral immunity experiment

[0076] Humoral immune antigen can specifically bind with the body produced, prevent it from infecting normal cells, and combine with macrophages, so that macrophages can phagocytize antigens to achieve the purpose of sterilization, so proteins with high humoral immune response intensity can also be Stimulate the body to produce antibodies to achieve immune protection.

[0077] 1. Experimental materials

[0078] Protein samples: Rv0237 protein and Rv1111c protein. Positive control 38kDa (Immuno Diagnostics Inc. USA).

[0079] Main reagents: mouse secondary antibody high-throughput kit, PS-MK15 Wes-Mouse 12-230 kDa master kit with split buffer for 200 data (Immuno Diagnostics Inc. USA). Streptomycin-labeled goat anti-human IgG, Thermo Scientific, Germany).

[0080] 2. Experimental principles and methods

[0081] Using Western Blot method. Western Blot technology is a classic protein detection technology. The traditional ...

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PUM

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Abstract

Belonging to the field of immunology and molecular biology, the invention relates to a tuberculosis immunodiagnosis molecular marker and application thereof in preparation of vaccines. Specifically, the invention relates to use of protein with an amino acid sequence shown as SEQ ID NO:1 or SEQ ID NO:2 in preparation of anti-tuberculosis drugs or mycobacterium tuberculosis-inhibiting drugs, or use in preparation of drugs detecting tuberculosis or drugs detecting mycobacterium tuberculosis. As an antigen, the protein shown as SEQ ID NO:1 or SEQ ID NO:2 has good sensitivity and specificity, also has reaction intensity significantly higher than that of positive control 38kDa, and has the potential of application to preparation of antituberculosis drugs like anti-tuberculosis vaccines.

Description

technical field [0001] The invention belongs to the field of immunology and molecular biology, and relates to a tuberculosis immunodiagnostic molecular marker and its application in preparing vaccines. Background technique [0002] Mycobacterium tuberculosis is one of the most threatening pathogens to human health. Tuberculosis is the main infectious disease among adults in the world today and has posed a serious challenge to international public health. After infecting the human body, Mycobacterium tuberculosis is mostly in a latent state or a persistent infection state. It can evade the host's defense in the infected host cells and reproduce in large numbers and achieve a delicate balance with the host. May be ill. Typically, 10% of infected people will develop active tuberculosis. An untreated active tuberculosis patient can infect 10-15 people a year. [0003] In the past 50 years, the theory and technology of tuberculosis control, clinical diagnosis and treatment lev...

Claims

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Application Information

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IPC IPC(8): A61K39/04A61P31/06C07K14/35C12N15/31
CPCA61K39/04A61K2039/53A61K2039/575C07K14/35
Inventor 金奇刘立国李海凤张笑冰张维佳
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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