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miRNA biomarkers, compositions and uses of the biomarker and the compositions

A biomarker and primer composition technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., can solve the problems of carcinogenesis, high false positive rate, lack of evidence, etc., to solve the problem of material collection , stable performance and convenient source

Inactive Publication Date: 2018-06-15
SHANGHAI LIDE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, the commonly used lung cancer screening method is low-dose spiral CT, but this technology also has obvious defects: (1) The false positive rate of screening is high (especially the pure ground-glass density shadow in the lung of 5-10mm) and the follow-up screening Long time, increased medical costs and the risk of radiation-induced cancer
(2) lead to excessive invasive biopsy and surgical intervention
[0004] So far, no miRNA with good specificity and sensitivity is suitable for use as a marker for lung cancer detection, especially serum markers, and there is a lack of relevant evidence

Method used

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  • miRNA biomarkers, compositions and uses of the biomarker and the compositions
  • miRNA biomarkers, compositions and uses of the biomarker and the compositions
  • miRNA biomarkers, compositions and uses of the biomarker and the compositions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 Identification of serum miRNA markers

[0086] (1) Sample collection

[0087] Sera from 10 lung cancer patients and 4 healthy people (normal controls) were collected;

[0088] (2) Extraction of RNA:

[0089] The miRNeasy Serum / Plasma Kit rapid extraction kit from Qiagen was used for RNA extraction. Each sample was loaded with 200 μl of plasma and the final elution volume was 50 μl. After the RNA sample is extracted, the quality of the sample is controlled by measuring the concentration and the ratio of OD260 / OD280. The optimal reaction result can be obtained when the ratio is between 1.8 and 2.0.

[0090] (3) PolyA tail:

[0091] Take 10μl extracted RNA sample, add 1x10 8 For each copy of cel-miRNA-39, add 4ul 5×PAP buffer, add 2-5U PolyA polymerase (Life Company, 74225Y / Z), and make up to 25μl with RNase-free water. Incubate at 37°C for 10-20 minutes, then at 65°C for 10 minutes.

[0092] (4) Reverse transcription reaction:

[0093] Take 10 μl of the ...

Embodiment 2

[0098] Example 2 The detection rate results of serum miRNA markers to lung cancer

[0099] The detection results of each serum miRNA marker (the above-mentioned expression difference is not less than 3 times is positive, and less than 3 times is negative) test results are compared with the "Chinese Common Malignant Tumor Diagnosis and Treatment Standards Sixth Volume Primary Disease" compiled by the Ministry of Health of my country. The sensitivity and specificity of lung cancer detection were calculated by comparing the results of the pathological diagnosis with the gold standard for the diagnosis of lung cancer in Bronchial Lung Cancer.

[0100] 1. Computational and statistical methods

[0101] The calculation method is as follows:

[0102]

[0103] The statistical analysis is as follows:

[0104] (1) Sensitivity: A / (A+C)

[0105] (2) Specificity: D / (B+D)

[0106] 2. Detection results of each miRNA

[0107] 1) miRNA-7:

[0108]

[0109]

[0110] Sensitivity: 80...

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Abstract

The invention relates to miRNA biomarkers, compositions and uses of the biomarker and the compositions. The biomarkers can be applied in preparation of lung cancer detection agents. The invention further relates to the composition comprising at least one of the miRNAs and sequences of a forward primer and a reverse primer of each miRNA, and uses of the composition in lung cancer detection agents.The identified miRNA is closely related to development of lung cancer, can be adopted as a biomarker for screening and diagnosing lung cancer, and provides an effective basis for clinical individual intervention treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular the invention relates to biomarkers, in particular to a serum miRNA biomarker and its application in lung cancer detection. Background technique [0002] Lung cancer is currently one of the malignant tumors with the highest morbidity and mortality in the world. 60-70% of patients are in advanced stage when they are first diagnosed, and the 5-year survival rate is only 16.1%. Up to 82%, so early screening is an important means to improve the survival rate of lung cancer. Currently, the commonly used lung cancer screening method is low-dose spiral CT, but this technology also has obvious defects: (1) The false positive rate of screening is high (especially the pure ground-glass density shadow in the lung of 5-10mm) and the follow-up screening A long time will increase the medical expenses and the risk of radiation-induced cancer. (2) lead to excessive invasive biopsy and surgical in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 闻丹忆李荣宇狄宇
Owner SHANGHAI LIDE BIOTECH CO LTD
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