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Type-1 diabetes markers and application thereof

A type 1 diabetes and marker technology, which is applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of not optimistic application prospects, lack of specificity, high cost, etc., and achieve good diagnosis Support efficiency, stable expression, make up for the effect of cumbersome detection

Inactive Publication Date: 2018-10-19
THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] C-peptide and islet autoantibodies such as glutamic acid decarboxylase antibody (GADA), zinc transporter 8 antibody (ZnT8A), islet cell antibody (ICA) are commonly used clinically to evaluate the degree of islet cell destruction and the prevalence of type 1 diabetes However, there are still many limitations in using it as a disease marker. For example, C-peptide can only reflect the characteristics of the disease in the late stage of islet destruction and lacks specificity; islet autoantibodies often appear successively, with intervals of several months or even tens In 2009, not all patients were positive for antibodies at the time of diagnosis, and many susceptible individuals who were positive for autoantibodies did not necessarily develop type 1 diabetes; these protein-based molecular tests were cumbersome, expensive, and time-consuming in clinical routine. The outlook is not optimistic

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  • Type-1 diabetes markers and application thereof
  • Type-1 diabetes markers and application thereof
  • Type-1 diabetes markers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Example 1 Screening of miRNA markers

[0131] The inventors used serum miRNA chip Agilent microarray (8*60k, 19.0) high-throughput chip technology (see the above-mentioned miRNA chip detection method for specific methods) to screen 6 cases of newly diagnosed typical T1DM patients and 6 cases of age-sex-matched healthy control serum For the expression of miRNA, up to 2006 miRNAs were detected.

[0132] It was found that 31 miRNAs were differentially expressed between the two groups. For details, see NCBI GeneExpression Omnibus (GSE94649, http: / / www.ncbi.nlm.nih.gov / geo).

[0133] According to the feasibility of the experiment, three miRNAs with up-regulated expression (hsa-miR-642a-3p, hsa-miR-320c, hsa-miR-1225-5p) and three miRNAs with down-regulated expression (hsa-let- 7b-5p, hsa-miR-26b-5p, hsa-miR-144-3p), a total of 6 miRNA enlarged sample verification, the corresponding nucleotide sequences are shown in Table 6 below:

[0134] Table 6 miRNA nucleotide sequence ...

Embodiment 21

[0141] Example 2 Auxiliary model for the diagnosis of type 1 diabetes

[0142] In order to judge the contribution value of each target miRNAs as a biomarker of T1DM (type 1 diabetes) in the diagnosis and prediction of T1DM and obtain the most effective diagnosis model, the inventors used Logistic regression analysis combined with each target miRNAs and clinical basic data to construct a diagnosis and prediction of T1DM model, hsa-miR-642a-3p, hsa-miR-320c, hsa-miR-1225-5p, hsa-let-7b-5p, hsa-miR-26b-5p , hsa-miR-144-3p these 6 target miRNA relative expression levels in the serum (2 -△Ct ) and age, sex, BMI into Logistic regression analysis.

[0143] Finally, the equation logit(P)=0.613+0.174×miR-320c(2 -△Ct )-2.199×miR-1225-5p(2 -△Ct )+0.09×age-0.213×BMI. The model was estimated using the maximum likelihood method, 2 = 37.75 (P value = 0.000). Among them, the four variables of hsa-miR-320c, hsa-miR-1225-5p, age, and BMI are left in the equation, which reflects the indepen...

Embodiment 31

[0144] Example 3 Stability and Reliability Test of Type 1 Diabetes Diagnosis Auxiliary Model

[0145] In order to verify the stability and reliability of the above-mentioned model, the inventor adopts leave-one-out method (leave-one-out) to carry out cross-validation (Cross-Validation) to the model on the training queue data. The raised function discriminates the case and calculates the misjudgment rate.

[0146] Finally, it was concluded that using miR-320c(2 -△Ct ), miR-1225-5p (2 -△Ct ), age, and BMI, the misjudgment rate of the model constructed by these four variables was 18.3%, which was consistent with the results obtained by the ROC analysis.

[0147]In order to further verify the diagnostic and predictive performance and reliability of the above model for T1DM, the inventor randomly collected serum samples from another independent verification cohort (33 cases of newly diagnosed T1DM and 29 cases of age-sex-matched healthy controls, the specific clinical characteris...

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Abstract

The invention discloses a set of type-1 diabetes markers, consisting of three miRNAs, including hsa-miR-642a-3p, hsa-miR-320c and hsa-miR-1225-5p; the invention also discloses the application of the markers in preparation of a type-1 diabetes diagnostic kit, and the kit contains reagents for quantifying the expression of the hsa-miR-642a-3p, hsa-miR-320c, hsa-miR-1225-5p. Compared with the known single biomarkers, utilizing a statistical method, and combining with multiple serum miRNAs and clinical demographic data to establish a type-1 diabetes serum miRNA-assisted diagnosis model, so the markers have better diagnostic support efficacy. The miRNA in circulation of a patient is utilized, the expression is stable, the storage and detection are easy, and clinical application is facilitated.

Description

technical field [0001] The invention relates to the technical field of type 1 diabetes diagnosis, in particular to a group of type 1 diabetes markers and applications thereof. Background technique [0002] At present, there is still a lack of effective disease markers for type 1 diabetes, and early screening and treatment are facing great challenges. There is an urgent need to find new disease markers to improve diagnostic and therapeutic performance. [0003] With the deepening of microRNA (microRNA, miRNA) research, its potential as a new non-invasive molecular marker has attracted more and more attention. Mature miRNA is a kind of endogenous small molecule single-stranded non-coding RNA composed of 21-23 nucleotides. By binding to the target mRNA base, it affects the translation or stability of mRNA, and regulates the gene at the post-transcriptional level. regulation. It has been confirmed that the human genome can encode more than 2,000 mature miRNAs, which regulate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 翁建平刘莉严晋华梁华
Owner THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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