Plasma exosome circRNA marker for non-small cell lung cancer as well as detection primer and kit thereof

A non-small cell lung cancer, marker technology, applied in the field of plasma exosome circRNA markers and its detection primers, kits, can solve the problems of poor specificity, low reliability, etc., to improve accuracy, broad application prospects, Ease of diagnosis

Active Publication Date: 2018-06-19
GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the problems of poor specificity and low reliability of existing non-small cell lung cancer detection and diagnosis markers, and provide a plasma exosomal circRNA marker for non-small cell lung cancer detection and diagnosis

Method used

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  • Plasma exosome circRNA marker for non-small cell lung cancer as well as detection primer and kit thereof
  • Plasma exosome circRNA marker for non-small cell lung cancer as well as detection primer and kit thereof
  • Plasma exosome circRNA marker for non-small cell lung cancer as well as detection primer and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Screening of markers for non-small cell lung cancer

[0045] 1. Research Methods

[0046] 1. Research population

[0047] The case group consisted of 245 cases of NSCLC collected from the Cancer Hospital Affiliated to Guangzhou Medical University, the First Affiliated Hospital of Guangzhou Medical University, the First People’s Hospital of Guangzhou, and the Chest Hospital of Guangzhou from November 2014 to December 2016. The diagnosis was confirmed histologically, and no surgery, radiotherapy or chemotherapy had been performed before blood collection. The control group consisted of patients with other lung diseases and healthy people in the physical examination center collected in the same period. According to the principle of the same gender, age ± 5 years, smoking status and similar smoking amount, 44 lung cancer cases were collected by frequency matching between groups. Patients with other diseases and 231 healthy controls. After signing the informed co...

Embodiment 2

[0055] Example 2 Establishment of non-small cell lung cancer diagnosis model and ROC analysis

[0056] The clinical value of the three plasma exosomal circRNAs obtained in Example 1 in the screening and diagnosis of NSCLC was analyzed by using the ROC curve method. The three circRNAs alone have significant diagnostic value for NSCLC. The accuracy of circ_0047921 is 0.577 (95% confidence interval is 0.523-0.631), the accuracy of circ_0007761 is 0.667 (95% confidence interval is 0.617-0.718), and the accuracy of circ_0056285 is 0.577 (95% confidence interval is 0.617-0.718). It reached 0.683 (95% confidence interval 0.634-0.732). The ROC curve analysis method of multivariate linear combination is used to combine the above four indicators, and the individual prediction probability model Y is obtained by binomial logistic regression pre1 =-40.106+0.131×Ct (circ_0047921) +0.663×Ct (circ_0056285) +417.86×1 / Ct (circ_0007761) . The diagnostic value of this combination for NSCLC...

Embodiment 3

[0058] Example 3 Preparation and clinical application of non-small cell lung cancer diagnostic kit

[0059] 1. Preparation of detection kits for clinical use

[0060] A diagnostic kit for non-small cell lung cancer, said kit comprising the following components: 1.5ml EP tubes, respectively equipped with specific primers for detecting the expression of circ_0047921, circ_0007761 and circ_0056285. The kit can also include various other reagents required for exosome isolation, RNA extraction, reverse transcription and fluorescent quantitative PCR detection, including but not limited to: Taq DNA polymerase, reverse transcriptase, dNTP Mixture, MgCl 2 solution, fluorescence quantitative PCR reaction buffer, deionized water and PBS buffer. The kit can be directly applied to clinic. The specific primer sequences are as follows:

[0061]circ_0047921-F: 5'-CAGCGCAGGAGAAAACGAAAC-3' (SEQ ID NO: 4);

[0062] circ_0047921-R: 5'-AGGCTGACAAGTGAGTGTGA-3' (SEQ ID NO: 5)

[0063] circ_000...

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Abstract

The invention discloses a plasma exosome circRNA marker for non-small cell lung cancer, as well as a detection primer and kit thereof. The circRNA marker is one or more of circ_0047921, circ_0007761,or circ_0056285. In a plasma exosome of an NSCLC patient, the expression level of the above-mentioned circRNAs is significantly different from that of plasma exosomes of other lung disease patients and healthy people. The invention also provides a specific primer group and a kit thereof for detecting circ_0047921, circ_0007761 and circ_ 0056285 respectively. The marker provided by the invention can be used alone in the diagnosis of NSCLC, and has certain accuracy, and a combination of the marker can significantly improve the accuracy of diagnosis (the AUC is 0.894, and the sensitivity and specificity are 87.25% and 80.00% respectively), and can differentiate NSCLC patients from other patients with lung diseases including pulmonary tuberculosis, chronic obstructive pulmonary disease and other disease patients, which is helpful to realize early detection of NSCLC.

Description

technical field [0001] The present invention relates to the field of biotechnology, more specifically, to a plasma exosomal circRNA marker of non-small cell lung cancer, detection primers and a kit thereof. Background technique [0002] Lung cancer is the most common malignant tumor in my country and even in the world, and its morbidity and mortality rate rank first among cancers. The latest data shows that in 2015, there were about 733,300 new cases of lung cancer among Chinese residents, accounting for 17.09% of all malignant tumors; the number of deaths reached 610,200, accounting for 21.68% of all malignant tumors. More than 80% of lung cancers are non-small cell lung cancer (NSCLC). The 5-year survival rate of NSCLC is still less than 20%. Studies have found that the 5-year survival rate (60%-80%) of lung cancer patients without metastasis is significantly higher than that of patients with local metastasis (survival rate 25%-50%) and distant metastasis (survival rate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 杨磊吕嘉春丘福满
Owner GUANGZHOU MEDICAL UNIV
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