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Molecular marker composition for diagnosing tuberculous pleurisy and application thereof

A technology of tuberculous pleurisy and molecular markers, applied in the field of medicine, can solve the problems that patients cannot obtain diagnosis results, cannot be detected, and limited diagnostic techniques

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

AI Technical Summary

Problems solved by technology

However, previous studies mainly focused on cancer, and some studies did not separate tuberculous pleurisy into a separate group, but combined it with other inflammatory diseases into a benign pleural disease group for comparative analysis with malignant pleuritic diseases
[0009] Clinically, the most direct diagnosis of tuberculous pleurisy is based on the presence of Mycobacterium tuberculosis in pleural effusion. However, due to limited diagnostic techniques, most suspected patients cannot obtain confirmed results. Mycobacterium tuberculosis found in 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 tuberculous pleurisy: 1. Tuberculosis detected in pleural effusion Mycobacterium, direct diagnosis of tuberculous pleurisy; 2. Mycobacterium tuberculosis was not detected in pleural effusion, but tuberculosis was detected in sputum, and tuberculous pleurisy was indirectly diagnosed; 3. Neither pleural effusion nor sputum was detected Mycobacterium tuberculosis can only be diagnosed clinically as tuberculous pleurisy by relying on other biochemical indicators and symptoms

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] miRNA microarray screening test

[0055] (1) Research objects

[0056] Patients with tuberculous pleurisy: Three categories of patients with tuberculous pleurisy were included according to the level of clinical evidence,

[0057] Group 1 (TPE1): patients with positive pleural effusion for Mycobacterium tuberculosis etiology;

[0058] Group 2 (TPE2): patients with pleural effusion negative for Mycobacterium tuberculosis pathogenic test but positive for sputum specimen pathogenic test;

[0059] Group 3 (TPE3): Both pleural effusion and sputum Mycobacterium tuberculosis pathogenic tests were negative, but the clinical diagnosis was tuberculous pleurisy.

[0060] Inclusion criteria for patients with tuberculous pleurisy:

[0061] ① There are clinical symptoms such as low fever, night sweats, weight loss, chest pain, dry cough; ② The pleural effusion is consistent with the change of exudate; ③ After anti-tuberculosis treatment, the pleural effusion is absorbed and the cli...

Embodiment 2

[0075] qPCR validation test

[0076] (1) Reagents:

[0077] Internal reference gene for qPCR: Cel-miR-39 (QIAGEN, Germany);

[0078] The reagents for RNA extraction, reverse transcription and qPCR detection were commercial reagents (QIAGEN, Germany).

[0079] (2) Primers:

[0080] miRNA upstream primer

[0081] Cel-miR-39: 5'-GCCGAGAGCTGATTTCGTCT-3'

[0082] hsa-miR-574-5p: 5'-TCGGCAGGTGAGTGTGTGT-3'

[0083] hsa-miR-4455: 5’-GCCGAGAGGGTGTGTGTGTT-3’

[0084] hsa-miR-4701-3p: 5’-TCGGCAGGATGGGTGATG-3’

[0085] The downstream primer is a universal primer: 5'-CTCAACTGGTGTCGTGGA-3'

[0086] (3) Instrument: ABI Quantistudio7.

[0087] (4) Test method:

[0088] The above 10 miRNAs were subjected to a large-scale qPCR validation test (188 patients with tuberculous pleurisy and 122 patients with malignant pleural disease).

[0089] The specific experimental process is as follows:

[0090] 1) RNA extraction: Take 250 μL of pleural effusion, centrifuge at 4°C, 16000×g for 10 mi...

Embodiment 3

[0106] Diagnostic Sensitivity and Specificity Analysis of ROC Analysis of Single miRNA

[0107] 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.

[0108] Table 4. Diagnostic value of a single miRNA in pleural effusion for differential diagnosis of tuberculous pleurisy and malignant pleural disease

[0109]

[0110] 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, which had a good differential diagnosis of tuberculous pleurisy and malignant pleural disease. Capacity for malignant pleural disease.

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Abstract

The invention relates to the technical field of medicines, in particular to a molecular marker composition for diagnosing tuberculous pleurisy and application of the molecular marker composition. The molecular marker composition is a composition of miR-4455 (micro Ribonucleic Acid), miR-574-5p (micro Ribonucleic Acid) and miR-4701-3p (micro Ribonucleic Acid). The molecular marker composition is used for preparing a diagnostic reagent or kit for tuberculous pleurisy. According to the invention, a high-throughput microarray chip is used for identifying pleural effusion differential miRNA for distinguishing tuberculous pleurisy and malignant pleural diseases for the first time, and further verification is carried out in an independent sample. The invention provides and identifies the miRNA composition which can be used for identifying and diagnosing focus parts of tuberculous pleurisy and malignant pleural diseases, and the miRNA composition is very important for improving the diagnosis of the tuberculous pleurisy.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a molecular marker composition for diagnosing tuberculous pleurisy and its application. Background technique [0002] A variety of diseases can cause abnormal accumulation of pleural effusion, including tuberculous pleurisy, malignant pleural disease, and pneumonia. Pleural effusions due to tuberculous pleurisy and malignant pleural disease are most common in countries with a high burden of TB. At present, the diagnosis of tuberculous pleurisy mainly relies on medical history, clinical manifestations, imaging, routine examination of pleural effusion and observation of curative effect, and the diagnosis rate is low, especially the differential diagnosis with malignant pleural disease and other infectious pleural diseases is difficult. [0003] At present, the gold standard for the diagnosis of tuberculous pleurisy is still the detection of Mycobacterium tuberculosis in pleural e...

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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