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Human circulating miR-122 detection method

A detection method and reverse transcription technology are applied in the detection field of human circulating miR-122, which can solve the problems of small research sample size and inconsistent results, and achieve the effect of improving sensitivity

Inactive Publication Date: 2016-10-26
SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES +1
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Problems solved by technology

However, the existing population studies that screen specific circulating miRNAs as biomarkers for obesity diagnosis and treatment have a small sample size, and the results reported by different studies are not consistent

Method used

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  • Human circulating miR-122 detection method
  • Human circulating miR-122 detection method
  • Human circulating miR-122 detection method

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Experimental program
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Embodiment 1

[0018] Study population: 18-30 years old population. Obesity patients are all patients in the obesity clinic; normal weight subjects are all school student volunteers.

[0019] 1. Serum RNA extraction

[0020] The total RNA of serum was extracted using the microRNA extraction kit (miRNeasy Mini Kit), and refer to the instructions provided by Qiagen, Germany.

[0021] (1) Take the serum sample out of the refrigerator at -80°C and place it on ice to completely melt;

[0022] (2) Add 1.5ml QIAzol Lysis Buffer to each EP tube containing serum samples, shake vigorously on the vortex mixer at the highest speed for 15s, and place on ice. Repeat this process until the white precipitate disappears completely , Let stand for 5min at room temperature;

[0023] (3) Add 200μl of chloroform solution to each tube, shake vigorously on the vortex mixer at the highest speed for 15s, and let it stand for 3min;

[0024] (4) Centrifuge at 12,000g for 15 minutes at 4°C, transfer the upper colorless water so...

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Abstract

The invention relates to a human circulating miR-122 detection method. The method includes: extracting serum total RNA (ribonucleic acid) as a sample, dissolving the sample into distilled water without nuclease, taking part of solution to perform reverse transcription to obtain cDNA, carrying out miR-122 specificity real-time fluorescent quantitative PCR (polymerase chain reaction) to obtain Ct values, and verifying chip reflected results. The human circulating miR-122 detection method has advantages that sensitivity is improved, a new clinical basis is provided for human circulating miR-122 variation under a condition of obesity, and circulating miR-122 level is closely related to obesity, glycoregulation and insulin resistance.

Description

Technical field [0001] The invention belongs to the field of obesity diagnostic biological markers, and particularly relates to a method for detecting human circulating miR-122. Background technique [0002] Studies have shown that fat cells play a central role in the pathogenesis of insulin resistance. When the body is obese, the secretory function of adipocytes is disordered, and the excessive accumulation of lipids leads to pathological processes such as cell lipotoxicity and insulin resistance. At present, basic research on obesity has initially revealed its pathogenesis, but biomarkers that can be used to diagnose obesity are still scarce, and the current treatment effects are not satisfactory. [0003] Recent basic studies have found that miRNAs play an important role in the occurrence and development of obesity and insulin resistance, but its regulatory mechanism has not yet been elucidated. In the fields of tumors, cardiovascular diseases and metabolic diseases, specific ...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6883C12Q2600/158C12Q2600/178C12Q2521/107C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 王卫庆王睿曹亚南洪洁顾卫琼张翼飞石娟
Owner SHANGHAI INST FOR ENDOCRINE & METABOLIC DISEASES
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