Molecular marker for auxiliary diagnosis of tubercular meningitis as well as application and kit thereof

A technology of tuberculous meningitis and molecular markers, which is applied in the field of disease diagnosis, can solve problems such as the inability to detect cerebrospinal fluid samples, and improve the effect of diagnosis

Pending Publication Date: 2022-04-19
BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the miRNAs that can be used for the detection of peripheral blood samples cannot be directly used for the detection of cerebrospinal fluid samples

Method used

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  • Molecular marker for auxiliary diagnosis of tubercular meningitis as well as application and kit thereof
  • Molecular marker for auxiliary diagnosis of tubercular meningitis as well as application and kit thereof
  • Molecular marker for auxiliary diagnosis of tubercular meningitis as well as application and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] In order to identify differential miRNAs for distinguishing tuberculous meningitis from other infectious meningitis, 11 patients with tuberculous meningitis and 12 patients with other infectious meningitis (including 7 patients with viral meningitis, 3 Genome-wide miRNA microarray analysis was performed on the cerebrospinal fluid of 1 patient with bacterial meningitis and 2 patients with cryptococcal meningitis, and then the original sample data was standardized to obtain the fold difference value (Fold Change), with Fold Change>1.5 and P< 0.05 screened out 7 differential miRNAs between the two groups. The results are shown in Table 1.

[0034] TBM (tuberculous meningitis) group: (1) The symptoms and signs of meningitis in patients include one or more of the following: headache, irritability, vomiting, fever, neck stiffness, convulsions, focal neurological dysfunction, altered consciousness or drowsiness. (2) Positive acid-fast staining of cerebrospinal fluid, or posi...

Embodiment 2

[0039] Real-time fluorescent quantitative PCR (qRT-PCR) to verify differential miRNA

[0040](1) Reagents: miRNA chip: Agilent human miRNA chip (8*60K) V21 chip; internal reference gene for qPCR: Cel-miR-39; reagents for RNA extraction, reverse transcription and qPCR detection are commercial reagents (QIAGEN).

[0041] (2) Internal reference gene for qPCR: Cel-miR-39. Internal reference and target primers: See the table below for the upstream primers, and the downstream primers are universal primers (downstream primer sequence: 5'-CTCAACTGGTGTCGTGGA-3')

[0042] miRNA upstream primer Cel-miR-39 5'-GCCGAGAGCTGATTTCGTCT-3' hsa-miR-21-5p 5'-GCCGAGTAGCTTTATCAGACTG-3' hsa-miR-1587 5'-TCGGCAGGTTGGGCTGGGCT-3' hsa-miR-3152-3p 5'-GCCGAGTGTGTTAGAATAGGG-3' hsa-miR-7975 5'-TCGGCAGGATCCTAGTCAC-3'

[0043] (3) Instrument: ABI Quantistudio7.

[0044] The results are shown in Table 2 and Table 3.

[0045] Table 2. qPCR detection raw data (...

Embodiment 3

[0050] The inventors tried to combine differentially expressed miRNAs to improve specificity and sensitivity.

[0051] Table 3. qPCR detection miRNA combination (predicted probability)

[0052]

[0053]

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Abstract

The invention relates to the technical field of disease diagnosis, in particular to a molecular marker for auxiliary diagnosis of tubercular meningitis as well as application and a kit thereof. The invention relates to a molecular marker for auxiliary diagnosis of tubercular meningitis. The molecular marker is miR-1587; the sequence of the hsa-miR-1587 is as shown in SEQ ID NO. 1 (sequence identifier number). The invention has the advantages that: (1) the cost is low; (2) the sensitivity is high; and (3) the specificity is good.

Description

technical field [0001] The invention relates to the technical field of disease diagnosis, in particular to a molecular marker for auxiliary diagnosis of tuberculous meningitis and its application and kit. Background technique [0002] The early clinical manifestations and imaging changes of patients with tuberculous meningitis are not specific, and the low sensitivity of current laboratory diagnostic tests makes the clinical diagnosis of tuberculous meningitis still obviously challenging. The detection of Mycobacterium tuberculosis in cerebrospinal fluid is the gold standard for the diagnosis of tuberculous meningitis, but the positive rate of Ziehl-Neelsen staining in traditional cerebrospinal fluid smears is low (10-20%), and the culture time of Mycobacterium tuberculosis in cerebrospinal fluid is long (6-8 weeks) and not sensitive enough (sensitivity ranged from 36% to 81.8%). [0003] The World Health Organization recommends the GeneXpert MTB / RIF test as a diagnosis in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/689C12Q1/6851C12N15/11C12R1/32
CPCC12Q1/6883C12Q1/689C12Q1/6851C12Q2600/178C12Q2600/158C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 潘丽萍谌蒙蒙董静史雨婷张宗德孙琦贾红彦魏荣荣杜博平邢爱英
Owner BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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