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MicroRNA 210 (MIR210) nucleic acid quantitative detection kit

A MIR210, detection kit technology, applied in the field of biotechnologists, can solve the problems of high false positive rate, only 1% positive rate, and complicated technical production

Inactive Publication Date: 2019-07-02
安徽普元生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the small number of tumors shed into the blood circulation, the sensitivity and specificity of this test are poor, and the positive rate is only 1%.
Later immunohistochemical methods have improved the detection rate, but the technology is complex to produce, requires specific antibodies, and has a high false positive rate

Method used

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  • MicroRNA 210 (MIR210) nucleic acid quantitative detection kit
  • MicroRNA 210 (MIR210) nucleic acid quantitative detection kit
  • MicroRNA 210 (MIR210) nucleic acid quantitative detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Application of microRNA 210 (MIR210) Nucleic Acid Quantitative Detection Kit (PCR-fluorescent probe method) to detect the expression of microRNA 210.

[0030] one. Material:

[0031] Sources of kit components and materials: RT-PCR buffer components in the one-step RT-PCR reaction solution were purchased from TAKARA Company, detection probes and primers were synthesized by TAKARA Company; DEPC-treated water was self-made; 2×10 6 The copies / µl standard is the plasmid solution of the target fragment; the positive control is the RNA sample with microRNA 210 (MIR210), and the negative control is the RNA sample without microRNA 210 (MIR210).

[0032] 2. Instruments and equipment:

[0033] ABI7300 fluorescent quantitative PCR instrument.

[0034] 3. Primer and probe design and synthesis:

[0035] Using the microRNA 210 (MIR210) sequence (GenBank accession number: NR 029623.1) as a template, use the random software of ABI 7300 real-time fluorescent quantitative PC...

Embodiment 2

[0044] Example 2 Clinical application of microRNA 210 (MIR210) nucleic acid quantitative detection kit (PCR-fluorescent probe method)

[0045] 1. Sample source

[0046] 50 patients with pathologically diagnosed lung cancer were included in the experimental group, of which 32 were pathologically diagnosed with lymph node or distant metastasis, 18 were pathologically diagnosed without lymph node or distant metastasis; 20 healthy people and 24 with benign lung diseases (10 cases of pneumonia, 8 cases of pulmonary tuberculosis, 6 cases of bronchitis) were the control group.

[0047] 2. Sample testing:

[0048] 1ml of serum was taken from each subject as a sample, and the RNA in the sample was extracted by an appropriate method. Fluorescent quantitative RT-PCR amplification is performed using the reagent of the present invention, and the RT-PCR reaction conditions are: 1) 30 minutes at 40°C; 2) 5 minutes at 95°C; 3) 15 seconds at 95°C → 1 minute at 60°C, 45 cycles. At the same t...

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Abstract

The invention relates to a microRNA 210 (MIR210) nucleic acid quantitative detection kit. RNA in a sample is subjected to reverse transcription by an RT-PCR method so as to obtain corresponding cDNA,and then a specific primers and probe of the kit are combined with a real-time fluorescence quantitative PCR detection technology so as to accurately detect the expression amount of microRNA 210 (MIR210) in the sample. The detection method can carry out quantitative analysis on microRNA 210 (MIR210) in peripheral blood serum samples of tumor patients, and can be clinically used for auxiliary diagnosis and evaluation of the therapeutic effect of prognosis. The kit guides clinical treatment through medical detection means in molecular science, and has certain clinical value.

Description

technical field [0001] The invention belongs to the field of biotechnologists, and is a kit for obtaining cDNA by reverse transcription (RT) RNA samples, combined with real-time fluorescent quantitative PCR technology, which can accurately and quantitatively detect the expression of human microRNA 210 (MIR210) in samples. Background technique [0002] According to data from the China Cancer Registry Center, in 2015, there were 4.3 million new cancer cases in China, and more than 2.81 million cancer deaths, accounting for 28.82% of the annual death toll, ranking first in the world, that is, more than 7,500 people died of cancer every day. cancer. Among all cancer deaths, the proportion of lung cancer deaths exceeds 35%, and the number of deaths has reached more than 500,000. On average, one out of every 15 deaths is a lung cancer patient. The survey shows that the incidence of lung cancer among urban residents in my country is close to that of developed countries. The overal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/178C12Q2561/113C12Q2563/107C12Q2561/101
Inventor 不公告发明人
Owner 安徽普元生物科技股份有限公司