Serum miRNA biomarker of type 2 diabetes mellitus and application thereof

A technology for diabetes and pre-diabetes, applied in the fields of biotechnology and medicine, can solve the problems of lack of effective means for type 2 diabetes

Inactive Publication Date: 2014-02-05
ZHEJIANG SCI-TECH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the relevant diagnostic criteria for type 2 diabetes include OGTT2h and HbA1c, there is still a lack of effective means for the early diagnosis and risk assessment of type 2 diabetes clinically, and sensitive and specific non-invasive serological indicators are needed. Especially non-invasive indicators based on miRNA

Method used

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  • Serum miRNA biomarker of type 2 diabetes mellitus and application thereof
  • Serum miRNA biomarker of type 2 diabetes mellitus and application thereof
  • Serum miRNA biomarker of type 2 diabetes mellitus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1. Collection of clinical samples and arrangement of clinical data.

[0037] The inventor obtained a large number of serum samples from Zhejiang Hospital of Traditional Chinese Medicine from November 2011 to March 2013, and systematically collected complete sample demographic and clinical examination data. The collected clinical samples were divided into three groups: normal glucose tolerance (NGT), prediabetes (IFG / IGT) and type 2 diabetes (T2D) according to WHO (1999) diagnostic criteria for diabetes (see Table 1). Refer to the case classification criteria of the WHO (1999) diagnostic criteria for diabetes (Table 1).

Embodiment 2

[0038] Example 2. Preliminary screening of type 2 diabetes serum-specific miRNA expression profile Solexa / Illumina high-throughput sequencing

[0039] (1) Screening of sequencing samples: By sorting out the sample data, the inventor selected 5 cases of type 2 diabetes (T2D) (50.60±5.13, age range 42-55, male: 3) from the collected clinical serum samples , female: 2) Serum samples were mixed thoroughly and used as mixed samples of type 2 diabetes case group (ie T2D group). In addition, 5 serum samples of normal glucose tolerance (NGT) (44.40±9.60, age range 34-58, male: 4, female: 1) were selected, mixed well, and used as the normal control group (NGT group) for Solexa / Experimental samples for primary screening by Illumina high-throughput sequencing.

[0040] (2) Use the Illumina TruSeq Small RNA Preparation kit kit to refer to the kit instructions Illumina’s TruSeq Small RNA Sample Preparation Guide to extract the total RNA of the T2D group and the NGT group respectively, an...

Embodiment 3

[0052] Example 3, Real-time fluorescence quantitative PCR method verification of type 2 diabetes serum candidate miRNA.

[0053] (1) Screening of samples: Through sorting out the sample data, the inventor selected 24 cases of type 2 diabetes (T2D) (51.13±9.21, age range 28-64, male: 16, Female: 8), 20 cases of prediabetes (IGT / FGT) (52.40±11.00, age span 31-65, male: 12, female: 8) and 20 cases of normal glucose tolerance (NGT) (46.65±16.18, age span 26 -75, male: 8, female: 12) as experimental samples for the verification of real-time fluorescent quantitative PCR method of serum candidate miRNA.

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Abstract

The invention discloses a serum miRNA composition, which is at least one of the following serum miRNAs: hsa-miR-23a, hsa-let-7i, and hsa-miR-486. The invention also simultaneously provides a fluorescent quantitative detection primer composition for detection of type 2 diabetes mellitus. The composition comprises an RT primer, an FW primer and an RV primer of each miRNA in hsa-miR-23a, hsa-let-7i, and hsa-miR-486. By adopting the serum miRNA marker for early diagnosis of type 2 diabetes mellitus and cancer risk evaluation provided by the invention, a serum miRNA biomarker can be used for diagnosing whether a subject has the type 2 diabetes mellitus or early diagnosis of the type 2 diabetes mellitus and cancer risk evaluation.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and relates to a serum miRNA-based biomarker and a method for detecting the expression amount thereof for the detection and early diagnosis of type 2 diabetes. Background technique [0002] Diabetes is an important endocrine and metabolic chronic disease. The number of diabetic patients is increasing rapidly around the world. It has become the third non-communicable disease that endangers human health after cardiovascular diseases and tumors. According to WHO recommendations, diabetes can be divided into type 1 diabetes, type 2 diabetes, specific diabetes and gestational diabetes. [0003] Type 2 diabetes, also called adult-onset diabetes, usually occurs after the age of 35 to 40, accounting for more than 90% of diabetic patients. Currently, indicators used for the diagnosis and classification of type 2 diabetes include: fasting plasma glucose (FPG), oral glucose tolerance test 2-hour ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12Q1/68
Inventor 陈海敏斯洪强李璇杨章萍丁倩芸丁先锋盛清
Owner ZHEJIANG SCI-TECH UNIV
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