MiRNA marker of immune thrombocytopenia, kit and application

A technology for thrombocytopenia and immunity, applied in the field of medical testing, to achieve high sensitivity, improve stability, and avoid data deviation

Inactive Publication Date: 2019-02-01
中国人民解放军第一00医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dysregulation of miRNA expression in cells can lead to the occurrence of various diseases such as immune thrombocytopenia. The latest research shows that some miRNAs are abnormally expressed in the peripheral blood of patients with immune thrombocytopenia, but which miRNA is related to the occurrence of immune thrombocytopenia There is still no consensus on the development

Method used

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  • MiRNA marker of immune thrombocytopenia, kit and application
  • MiRNA marker of immune thrombocytopenia, kit and application
  • MiRNA marker of immune thrombocytopenia, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1 The method for detecting miRNA markers of immune thrombocytopenia

[0040] 1. Collection of clinical samples

[0041]The inventor has collected a large number of patients with immune thrombocytopenia from the 100th Hospital of the People's Liberation Army and the Affiliated Hospital of Soochow University since 2016. The diagnostic criteria all meet the "Chinese Expert Consensus on the Diagnosis and Treatment of Primary Immune Thrombocytopenia in Adults (Revised Edition)"; At the same time, the healthy positive control group in the physical examination center was selected, and peripheral blood was drawn. This study was approved by the Hospital Ethics Committee, and all subjects signed informed consent. The process of collection and follow-up experiments complied with the requirements of medical ethics and strictly followed the principle of confidentiality of case data. The sampling, packaging and storage conditions of the research samples were the same. By a...

Embodiment 2

[0105] This implementation provides an auxiliary diagnostic kit for immune thrombocytopenia, which is used to detect the expression level of hsa-miR-302e in peripheral blood. The kit contains miRNA markers of immune thrombocytopenia; the base sequence of the miRNA markers is shown in SEQ ID NO:1. The kit also includes primers for miRNA markers, the base sequences of which are shown in SEQ ID NO: 2 and SEQ ID NO: 3. The kit also includes commonly used reagents for total RNA extraction, cDNA reverse transcription primers, commonly used reagents for reverse transcription, polymerase and reagents commonly used for fluorescent quantitative Q-PCR reactions and fluorescent detection reagents (see Example 1); the cDNA The reverse transcription primer sequence is shown in SEQ ID NO:4. The kit also includes a standard and / or a reference; the standard and / or a reference is endogenous miRNA-U6RNA.

[0106] Using the kit can achieve the same detection effect as the embodiment, and the op...

Embodiment 3

[0108] ROC curve was used to analyze the diagnostic value of hsa-miR-302e level for ITP. Among the 127 test subjects, with ITP patients as the target subjects, the ROC curve was drawn using Graphpad software, and various related parameters were analyzed. Such as Figure 4 As shown, the area under the curve reaches 0.8913, indicating that it has a very good diagnostic value. Using the principle of the largest Youden's index, the optimal cut-off value of hsa-miR-302e was judged to be 1.552, and the corresponding sensitivity and specificity were 77.54 and 95%, respectively, as shown in Table 2.

[0109] Table 2:

[0110]

[0111]

[0112] Depend on Figure 4 It can be seen from Table 2 that the area under the curve is 0.0.891±0.027 (P<0.001), suggesting that it has a high diagnostic value. The optimal cut-off value of hsa-miR-302e concentration was 1.552, and the corresponding sensitivity and specificity were 77.53% and 95%, respectively.

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Abstract

The invention provides a miRNA marker of immune thrombocytopenia, a kit and application. The miRNA marker is hsa-miR-302e, the base sequence of the marker is shown as SEQ ID NO:1. The base sequences of the miRNA marker's primers are shown as SEQ ID NO:2 and SEQ ID NO:3. The miRNA marker and the primers thereof can be used for preparation of drugs inhibiting immune thrombocytopenia or preparation of immune thrombocytopenia auxiliary diagnosis kit. The miRNA marker can be used for judging the progress and severity of immune thrombocytopenia. The hsa-miR-302e primers, diagnosis kit and detectionmethod developed based on the miRNA marker have the characteristics of simple operation, little trauma to examinees, high sensitivity and strong specificity, and can be used for diagnosis of immune thrombocytopenia at the early stage and different stages, and have obvious clinical application value.

Description

technical field [0001] The invention belongs to the technical field of medical testing, and in particular relates to a miRNA marker for immune thrombocytopenia, a kit and an application thereof. Background technique [0002] MicroRNA (miRNA) is a small non-coding RNA with a length of about 19-23nt that exists in the body. It binds to the complementary sequence in the 5'-UTR or 3'-UTR of the target gene mRNA through the base pairing principle to degrade the mRNA Or inhibit the translation of related proteins, thereby regulating gene expression at the post-transcriptional level. One microRNA may bind to multiple mRNAs, and one mRNA may also be regulated by multiple microRNAs. About 1 / 3 of genes in living organisms are regulated by miRNAs. However, there is still controversy about which miRNAs are abnormally expressed in ITP patients, and the specific mechanism of miRNAs in the occurrence and development of ITP has not yet been elucidated. [0003] Primary immune thrombocytop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113C12N15/11
CPCC12N15/113C12N2310/141C12Q1/6883C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 钱琤吴天勤任传路邓安梅严文颖沈红石李征洋
Owner 中国人民解放军第一00医院
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