Molecular marker for diagnosing tuberculous pleurisy and application thereof

A technology of tuberculous pleurisy and molecular markers, applied in the field of medicine, can solve problems such as limited diagnostic techniques, failure to detect, and inability to obtain diagnosis results for patients, and achieve good identification effect, high accuracy, and high specificity Effect

Pending Publication Date: 2022-07-01
BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Clinically, the most direct diagnosis of tuberculous pleurisy is based on the presence of Mycobacterium tuberculosis in pleural effusion, but due to limited diagnostic techniques, most suspected patients cannot obtain confirmed results. Mycobacterium tuberculosis found in the pleural effusion), other biochemical indicators, and clinical symptoms, etc. Therefore, clinically, according to the level of diagnostic evidence, it is usually divided into three types of tuber

Method used

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  • Molecular marker for diagnosing tuberculous pleurisy and application thereof
  • Molecular marker for diagnosing tuberculous pleurisy and application thereof
  • Molecular marker for diagnosing tuberculous pleurisy and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] miRNA microarray screening test

[0069] (1) Research objects

[0070] Patients with tuberculous pleurisy: Three categories of patients with tuberculous pleurisy were included according to the level of clinical evidence,

[0071] Group 1 (TPE1): patients with positive pleural effusion for Mycobacterium tuberculosis etiology;

[0072] Group 2 (TPE2): patients with pleural effusion negative for Mycobacterium tuberculosis pathogenic test but positive for sputum specimen pathogenic test;

[0073] Group 3 (TPE3): Both pleural effusion and sputum Mycobacterium tuberculosis pathogenic tests were negative, but the clinical diagnosis was tuberculous pleurisy.

[0074] Inclusion criteria for patients with tuberculous pleurisy:

[0075] ①Clinical symptoms such as low fever, night sweats, weight loss, chest pain and dry cough;

[0076] ②The pleural effusion is consistent with the change of exudate;

[0077] ③ After anti-tuberculosis treatment, the pleural effusion was absorbed...

Embodiment 2

[0097] qPCR validation test

[0098] (1) Reagents:

[0099] Internal reference gene for qPCR: Cel-miR-39 (QIAGEN, Germany);

[0100] The reagents for RNA extraction, reverse transcription and qPCR detection were commercial reagents (QIAGEN, Germany).

[0101] (2) Primers:

[0102] miRNA upstream primer

[0103] Cel-miR-39: 5'-GCCGAGAGCTGATTTCGTCT-3'

[0104] hsa-miR-574-5p: 5'-TCGGCAGGTGAGTGTGTGT-3'

[0105] hsa-miR-4455: 5’-GCCGAGAGGGTGTGTGTGTT-3’

[0106] hsa-miR-4701-3p: 5’-TCGGCAGGATGGGTGATG-3’

[0107] The downstream primer is a universal primer: 5'-CTCAACTGGTGTCGTGGA-3'

[0108] (3) Instrument: ABI Quantistudio7.

[0109] (4) Test method:

[0110] The above 10 miRNAs were subjected to a large-sample qPCR validation test (188 patients with tuberculous pleurisy and 122 patients with malignant pleural disease). The specific experimental process is as follows:

[0111] 1) RNA extraction: Take 250 μL of pleural effusion, centrifuge at 16000×g at 4°C for 10 min, draw ...

Embodiment 3

[0128] Diagnostic Sensitivity and Specificity Analysis of ROC Analysis of Single miRNA

[0129] Three differential miRNAs, miR-574-5p, miR-4701-3p, and miR-4455 in pleural effusion, were analyzed using receiver operating characteristic (ROC) curves in 188 patients with tuberculous pleurisy and 122 patients with malignant pleural disease. The ROC curve analysis was carried out, and the results are shown in Table 4.

[0130] The area under the ROC curve is Figures 1 to 3 shown.

[0131] Table 4. Diagnostic value of a single miRNA in pleural effusion for differential diagnosis of tuberculous pleurisy and malignant pleural disease

[0132]

[0133] The results showed that the area under the ROC curve (AUC) of miR-574-5p, miR-4701-3p and miR-4455 for the differential diagnosis of tuberculous pleurisy and malignant pleural disease were all greater than 0.9, and pleural effusion miR-574-5p had Ability to differentiate between tuberculous pleurisy and malignant pleural disease....

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Abstract

The invention relates to the technical field of medicines, in particular to a molecular marker for diagnosing tuberculous pleurisy and application of the molecular marker. The molecular marker is miR-4455 (micro Ribonucleic Acid) or miR-574-5p (micro Ribonucleic Acid) or miR-4701-3p (micro Ribonucleic Acid). The molecular marker is used for preparing a diagnostic reagent or kit for tuberculous pleurisy. The method has the advantages that (1) the identification effect is good; (2) the sensitivity is high; and (3) the specificity is high. And (4) the method is suitable for diagnosis and has high accuracy.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a molecular marker for diagnosing tuberculous pleurisy and its application. Background technique [0002] There are about 5-15ml of fluid in the thoracic cavity of normal people to separate the two layers of pleura, which plays a lubricating role during breathing. The volume of pleural fluid is not fixed, and normal people also have 500-1000ml of fluid exudation and reabsorption every 24 hours, and the two are in a state of balance. Pleural effusion occurs when there is increased fluid in the pleural space due to increased exudation and / or decreased reabsorption for any reason. Pleural effusion is divided into two categories: exudate and exudate according to the etiology and biochemical properties, which can be clinically identified by Li Fanta experiment. But in the category of exudative pleural effusion, among many diseases that can cause pleural effusion, tuberculous pleuri...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178C12Q2600/112
Inventor 潘丽萍董静张宗德史雨婷贾红彦孙琦李自慧杜博平魏荣荣邢爱英
Owner BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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