miRNA marker for assistant diagnosis of tuberculosis and application thereof

A diagnostic marker and auxiliary diagnosis technology, which can be used in the determination/examination of microorganisms, biochemical equipment and methods, etc., and can solve the problems of complex components and unsatisfactory effects.

Pending Publication Date: 2019-05-31
BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the composition of serum and plasma is complex, and the real specific molecular markers may be trace...

Method used

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  • miRNA marker for assistant diagnosis of tuberculosis and application thereof
  • miRNA marker for assistant diagnosis of tuberculosis and application thereof
  • miRNA marker for assistant diagnosis of tuberculosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1. Discovery of miRNAs that can be used as diagnostic markers

[0064] The healthy cohort (HC) consisted of 60 healthy subjects. The tuberculosis latent infection cohort (LTBI) consisted of 60 cases of latent tuberculosis infection. The active tuberculosis cohort (ATB) consisted of 60 patients with active tuberculosis.

[0065] 1. Prepare the sample pool

[0066] The serum of 60 healthy persons in the healthy group was collected respectively, and the serum was mixed (1 ml / case) to obtain a sample pool of the healthy group. The serum of 60 cases of latent tuberculosis infection in the latent tuberculosis infection group was collected respectively, and the serum was mixed (1ml / case) to obtain the sample pool of the latent tuberculosis infection group. The serum of 60 patients with active pulmonary tuberculosis in the active pulmonary tuberculosis population group was collected respectively, and the serum was mixed (1 ml / case) to obtain a sample pool of the acti...

Embodiment 2

[0091] Example 2, further screening of miRNAs that can be used as diagnostic markers from 20 miRNAs

[0092] Subjects: 10 healthy persons, 10 latent tuberculosis infection patients, and 10 active pulmonary tuberculosis patients.

[0093] 1. Extract exosomes

[0094] Serum exosomes were extracted from each subject separately.

[0095] 1. Take the subject's serum, centrifuge at 1000g for 15min, and collect the supernatant.

[0096] 2. Take the supernatant obtained in step 1, centrifuge at 16500 g for 30 min at 4° C., and collect the supernatant.

[0097] 3. Take the supernatant obtained in step 2, filter it with a filter membrane with a pore size of 0.22 μm, and collect the filtrate.

[0098] 4. Take the filtrate obtained in step 3, centrifuge at 4°C and 120,000 g for 3 hours, discard the supernatant, turn the centrifuge tube upside down on absorbent paper to drain the liquid on the tube wall, and the precipitate attached to the bottom of the centrifuge tube is exosomes prec...

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Abstract

The invention discloses a miRNA marker for assistant diagnosis of tuberculosis and application thereof. The invention requires the protection of the application of a substance for detecting miRNAs inserum exosomes in the preparation of products used for diagnosing or assisting the diagnosis of tuberculosis patients or for diagnosing or assisting the diagnosis of active tuberculosis patients, wherein each miRNA refers to one or a combination of the following five kinds of miRNAs: miR-28-3p, miR-193b-5p, miR-370-3p, miR-1246 and miR-2110. The invention also requires the application of the substance for detecting miRNAs in serum exosomes in the preparation of products used for diagnosing or assisting the diagnosis of patients with latent tuberculosis infections, wherein each miRNA refers toone or a combination of the following three kinds of miRNAs: let-7e-5p, let-7d-5p and miR-140-5p. A foundation is laid for further development of the early rapid diagnosis technology and method for tuberculosis.

Description

technical field [0001] The present invention relates to miRNA markers for auxiliary diagnosis of tuberculosis and applications thereof, in particular to miRNA markers for auxiliary diagnosis of tuberculosis in serum exosomes and applications thereof. Background technique [0002] The current gold standard for TB diagnosis is bacteriological sputum smear microscopy and sputum culture. Bacteriological detection has low sensitivity and takes a long time, which is not conducive to the diagnosis and treatment of various stages of tuberculosis. People susceptible to tuberculosis (such as children) are often unable to cooperate with coughing up sputum, and it is difficult to collect sputum samples. The limitations of bacteriological detection are more obvious. It is necessary to look for diagnostic specimens other than sputum, such as easily collected blood. However, the composition of serum and plasma is complex, and the real specific molecular markers may be trace, which leads...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
Inventor 吕翎娜张秀丽李翠丹杨婷婷张宗德陈非潘丽萍李自慧
Owner BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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