Application of ASPH gene in exosome in lung cancer early diagnosis kit

An early diagnosis and exosome technology, applied in the field of genetic testing, can solve the problems of high false positive, expensive and dangerous

Inactive Publication Date: 2020-06-12
江西惠肽生物科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Screening asymptomatic patients with CT scans is expensive and dangerous
This screening has a high rate of false positives and some of these patients will receive unnecessary lung biopsies
At present, there are no related reports on the method of detecting ASPH gene in exosomes

Method used

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  • Application of ASPH gene in exosome in lung cancer early diagnosis kit
  • Application of ASPH gene in exosome in lung cancer early diagnosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Design primers according to the ASPH gene, and the obtained primer sequences are as follows:

[0031] ASPH-F: 5'-AGCGGTAGCACGAGTGC-3'; SEQ ID NO.1;

[0032] ASPH-R: 5'-GCCCAGCAATGCAATCAC-3'; SEQ ID NO.2.

[0033] (2) Extract exosomes from lung cancer tumor cells (A549, NCI-H1299, NCI-H1650, HCC827) and normal bronchial cells (NL-20) culture supernatant; add 1ml PBS to dilute 1ml supernatant, then add 1ml Exosome extraction reagents to obtain exosomes.

[0034] (3) RNA extraction: the obtained exosomes were dissolved in 700 μl Qiazol to extract RNA, and the quality and quantity of RNA were determined.

[0035] (4) Using the SYB Green method and the primers of step (1), carry out RT-qPCR analysis on an ABI PCR instrument.

[0036] β-actin was used as an internal reference, and the primer sequences were as follows:

[0037] β-actin-F: 5'-TGGCATTGCCGACAGGAT-3'; SEQ ID NO.3;

[0038] β-actin-R: 5'-ATACTCCTGCTTGCTGATCCACAT-3'; SEQ ID NO.4.

[0039] The reaction syst...

Embodiment 2

[0042] Embodiment 2 Utilizes ROC curve to detect the specificity and the sensitivity of primer

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Abstract

The invention discloses application of ASPH gene in an exosome in a lung cancer early diagnosis kit, and belongs to the technical field of gene detecting. The invention discloses the application of the ASPH gene in the exosome in the lung cancer early diagnosis kit. The kit comprises a primer for detecting the ASPH gene in the exosome and a primer for detecting beta-actin. The application of the ASPH gene in the exosome in the lung cancer early diagnosis kit disclosed by the invention is high in detecting sensitivity and strong in specificity.

Description

technical field [0001] The present invention relates to the technical field of gene detection, and more specifically relates to the application of the ASPH gene in exosomes in a lung cancer early diagnosis kit. Background technique [0002] The World Health Organization recently released the "World Cancer Report"; the report said that the incidence of cancer in mainland China accounted for almost half of the world's total. The report predicts that by 2030, the number of cancer patients worldwide will increase by 50%. Among the global cancer cases, lung cancer has the fastest growth rate, reaching 1.8 million people, and lung cancer has risen to the number one malignant tumor with the highest incidence. Lung cancer is the cancer with the highest morbidity and mortality rate in the world. In 2012 alone, 1.6 million people died of lung cancer. This type of cancer is difficult to treat, and metastatic lung cancer is particularly difficult to treat. About 75% of lung cancer pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6851C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 余跃飞刘怀朱皓皞
Owner 江西惠肽生物科技有限公司
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