High-specificity lnc RNA biomarker for heart failure and detection method and application thereof

A biomarker and specific technology, applied in the field of heart failure lncRNA biomarkers and its detection, new biomarkers to evaluate heart failure, can solve the problems of poor repeatability and specificity of results, and achieve great theoretical significance

Active Publication Date: 2020-03-13
南京启医科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although various research results have shown that the combination of multiple miRNAs can improve the prediction performance, the reproducibility of the results of different researchers is poor. Since miRNAs are involved in the expression regulation of most genes, the different miRNA expression profiles observed in heart failure are only partially Reflects molecular pathology associated with heart failure, relatively poor specificity

Method used

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  • High-specificity lnc RNA biomarker for heart failure and detection method and application thereof
  • High-specificity lnc RNA biomarker for heart failure and detection method and application thereof
  • High-specificity lnc RNA biomarker for heart failure and detection method and application thereof

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Effect test

Embodiment 1

[0022] Fluorescent quantitative PCR method was used to detect the content of lncRBA-B and lncRNA-C in the male C57BL heart failure mouse model, and the specific steps were as follows:

[0023] Step 1, extracting lncRBA-B and lncRNA-C

[0024] 1. Acute HF mouse model can be induced by subcutaneous injection of isoproterenol (40mg / kg / day) for 1 week.

[0025] 2. Peripheral whole blood was collected through the orbital vein.

[0026] 3. Use the whole blood RNA extraction kit to extract the total RNA of the whole blood sample with the phenol extraction method according to the instructions. Plasma was obtained by centrifuging EDTA-blood at 1600 g for 10 min, and the supernatant was centrifuged at 12000 g for 5 min at 4°C. Clean plasma was filtered through a 0.22 μm filter and mixed with 2 mL of TRIZOL reagent and 0.4 mL of chloroform. The mixture was centrifuged at 12000 g for 5 min at 4 °C, then the aqueous layer was transferred to a new tube, 1.5 mL of 70% ethanol was added, a...

Embodiment 2

[0044] Detection of lncRBA-B and lncRNA-C in patients with coronary heart disease with heart failure

[0045] Specific steps are as follows:

[0046] Step 1, extract total RNA from serum

[0047] 1. Serum was centrifuged at 10,000×g for 30 minutes, and the supernatant was centrifuged with a low-temperature ultra-high-speed centrifuge L-80XP 110,000×g for 70 minutes to obtain a precipitate. Add PBS to resuspend and mix, and then repeat ultra-high-speed centrifugation once to obtain exosomes. Resuspend the pellet.

[0048] 2. Take 20-40 μl exosome resuspension and drop it on the special copper grid for electron microscope, 20 μl 2% phosphotungstic acid negative staining for 10 minutes, put it under a transmission electron microscope H-7650 to observe the shape of exosomes and take pictures.

[0049] 3. Use with ultra-high-speed centrifugation Take 750 μl TRIzol-LS to lyse 250 μl exosome suspension, then use chloroform to separate the lysate, take the upper layer and add isopro...

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Abstract

The invention relates to a high-specificity lnc RNA biomarker for heart failure and a detection method and application thereof. The biomarker is lnc RNA-B and lnc RNA-C. The expression level of lnc RNA-B and lnc RNA-C is correlated with the development of heart failure. The expression level of lnc RNA-B and lnc RNA-C is quantified to diagnose whether a subject suffers from heart failure or has a higher risk of heart failure. The sequence of Inc RNA-B is SEQ NO.1, and the sequence of lnc RNA-C is SEQ NO.2. The lnc RNA-B and lnc RNA-C are both isolated from peripheral venous blood plasma. The lnc RNA biomarker can be used for preparing a heart failure diagnostic reagent or a biochip; and real-time fluorescence quantitative PCR is adopted to detect expression level of lnc RNA-B and expressionlevel of lnc RNA-C respectively. The heart failure biomarker in the invention makes up for the inapplicability of some other biomarkers, provides help for diagnosing heart failure and reflects different processes of heart failure, is more sensitive and accurate than other existing heart failure biomarkers, and has great theoretical significance and potential use value.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a lnc RNA biomarker of heart failure with high specificity and its detection method and application, and the biomarker of heart failure with high specificity is evaluated as a new type of biological index heart failure. Background technique [0002] Heart failure is a complex clinical syndrome, a severe stage of various heart diseases, and the leading cause of death from cardiovascular diseases. In 2003, the prevalence of heart failure in my country was 0.9%, and there were more than 4 million patients with heart failure. Due to the rising incidence of hypertension, coronary heart disease, and diabetes in my country, and the serious aging population, the prevalence of heart failure continued to rise Unavoidable, the five-year survival rate of patients with heart failure is similar to that of malignant tumors. Patients with heart failure manifest early symptoms of fatigue, ch...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/178Y02A50/30
Inventor 董鸣杨子仪徐聪周彦清张锐
Owner 南京启医科技有限公司
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