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Molecular markers for auxiliary diagnosis of viral meningitis and their applications and kits

A technology of viral meningitis and molecular markers, applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc., can solve problems such as confusion of other diseases, low positive detection rate, and difficulty in finding lesions , to achieve the effect of improving early diagnosis rate, reducing mortality rate and high diagnostic value

Active Publication Date: 2022-06-21
THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the process of meningitis is often limited to the central and deep layers of the temporal lobe. If the materials are not obtained properly, the purpose will not be achieved. Moreover, there are many types of viruses, and the number of new or unexplained viral meningitis is increasing. It is difficult to isolate viruses routinely. And there is a lack of specific and sensitive molecular biological detection methods
①Using brain tissue for virus isolation is currently recognized as the most reliable method for diagnosis, but it cannot be used as a routine diagnosis method. The first is the time of diagnosis. Some viruses (such as herpes simplex virus) can disappear from the nerve tissue in a short time. Secondly, biopsy May also cause damage to the brain, occasionally causing massive bleeding or concurrent infection
③ Detection of virus genes in cerebrospinal fluid by PCR technology is an emerging molecular gene detection technology. The detection method is fast and convenient. However, the content of virus or its antigen in cerebrospinal fluid CSF is very small, resulting in a very low positive rate of detection, prone to false positives, and relies on genome sequencing platforms
④ Immunological and serological methods also have certain limitations, and limited by laboratory conditions, they have not been widely carried out, and the diagnosis is often non-specific
[0006] (3) Imaging techniques such as CT and MRI have a very low positive rate in the early diagnosis of viral meningitis. Generally, it is difficult to find lesions or have no abnormal manifestations in the early stage. The EEG wave pattern of viral meningitis is mainly diffuse Irregular high-amplitude slow waves. For focal lesions, the EEG usually shows that one side is heavier, which can help to judge the lesion site. In patients with viral meningitis, brain function abnormalities are earlier than structural abnormalities, and the definition of the lesion edge is not clear. First, it is easy to cause confusion with other diseases, and in the early stage of the onset, its imaging specificity is not obvious. The accuracy of CT, MRI and other imaging techniques for the diagnosis of viral meningitis is mainly in the middle and early stages of the onset. Late stage, and its examination is expensive, not suitable as a routine screening method for the general population

Method used

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  • Molecular markers for auxiliary diagnosis of viral meningitis and their applications and kits
  • Molecular markers for auxiliary diagnosis of viral meningitis and their applications and kits
  • Molecular markers for auxiliary diagnosis of viral meningitis and their applications and kits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1: density and boxplot analysis results

[0080] By extracting small RNAs from the cerebrospinal fluid of patients with viral meningitis infection and healthy adults, miRNA identification was performed, and the bioinformatics analysis statistics were density and boxplot analysis graphs ( figure 2 ), it was found that there was no difference in the overall number of miRNAs in the diseased brain group and the control group.

Embodiment 2

[0081] Embodiment 2: density and boxplot analysis graph PCA and volcano plot analysis result graph

[0082] By extracting small RNAs from the cerebrospinal fluid of patients with viral meningitis infection, patients with non-viral infection, and healthy adults, miRNA identification was performed, and the bioinformatics analysis and statistics were PCA and volcano plot analysis graphs (such as image 3 ), it was found that the overall number of miRNAs in the diseased brain group and the control group was almost the same.

Embodiment 3

[0083] Example 3 Heatmap

[0084] By extracting small RNAs from the cerebrospinal fluid of patients with viral meningitis infection and healthy adults, miRNA identification was performed, and the differential expression of differentially expressed genes in different samples was visualized by Heatmap (such as Figure 4 ).

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Abstract

The invention discloses a molecular marker for auxiliary clinical diagnosis of viral meningitis, its application and a kit. Firstly, the miSeq high-throughput sequencing technology was used to analyze the two groups of cerebrospinal fluid samples of viral meningitis and normal controls, and the differential miRNA expression was screened, and then the RT-qPCR technology was used to verify the differentially expressed miRNAs one by one, and finally found that , miR‑RNA 18_41385, hsa‑miR‑486‑5p, hsa‑miR‑4497 and hsa‑miR‑3178 can be used as specific markers for early auxiliary clinical diagnosis of viral meningitis. These markers also enable differential diagnosis of other meningitis. The kit of the present invention has the advantages of good stability, simple and convenient detection method, high sensitivity and specificity, and realizes an effective supplement to existing clinical viral meningitis diagnostic methods and rapid diagnosis of viral meningitis patients. It is of great significance to reduce the mortality rate of viral meningitis, improve the treatment effect and prognosis.

Description

technical field [0001] The invention belongs to the technical field of diagnosis of viral meningitis, in particular to a cerebrospinal fluid-specific molecular marker for diagnosis of viral meningitis, its application and a matching detection kit. Background technique [0002] In our country and even the whole world, viral meningitis is a relatively common and serious infectious disease of the central nervous system, which can be caused by various viruses invading the meninges. Viral meningitis can cause fever, headache, vomiting, and in severe cases, permanent brain damage and even death. The clinical manifestations are very similar to other types of central nervous system infections, which need to be identified by combining medical history, cerebrospinal fluid examination and etiological analysis; at the same time, some non-viral infectious diseases such as purulent meningitis, fungal meningitis, tuberculous meningitis, etc. The symptoms are also similar to those of viral...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/178C12Q2600/158
Inventor 朱海霞孙嘉萌曾赵军唐潇蒙罗兰
Owner THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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