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Kit, device and method for lung cancer diagnosis

A lung cancer diagnosis and kit technology, applied in the field of medical diagnosis, can solve the problem of low sensitivity

Active Publication Date: 2020-07-07
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, not all pulmonary nodules are malignant, and the distinction between benign and malignant pulmonary nodules has always been a difficult point in clinical diagnosis and treatment in thoracic surgery
At present, there are also non-invasive detection methods using plasma circulating tumor cells and circulating tumor cell-free DNA, etc., but their detection sensitivity in the diagnosis of early lung cancer is not high; therefore, it is urgent to develop a high-sensitivity non-invasive early detection method for lung cancer

Method used

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  • Kit, device and method for lung cancer diagnosis
  • Kit, device and method for lung cancer diagnosis
  • Kit, device and method for lung cancer diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1 Screening of exosomal mRNA and LncRNA markers associated with early lung cancer based on high-throughput sequencing

[0093] In order to screen for exosome markers associated with the diagnosis of early lung cancer, 72 patients with early lung cancer diagnosis and 50 controls each took no less than 10ml of blood and separated the plasma, which was used to separate the exosomes in the plasma by classical ultracentrifugation. The qiagen miRNeasy mini kit was used to extract RNA, and the obtained RNA was subjected to a small amount of ribosomal chain-specific RNA library construction and sequencing. The obtained data was analyzed by bioinformatics, and the differentially expressed mRNA and LncRNA (collectively referred to as long RNA) were compared between patients with early lung cancer and controls, and the long RNAs with significant differences were obtained as shown in Table 2 below. These long RNA-level markers from exosomes can be used for early diagnosis o...

Embodiment 2

[0096] Embodiment 2 mRNA and LncRNA detection system based on fluorescent quantitative PCR platform

[0097] 1. Reverse transcription of mRNA and LncRNA

[0098] Using PrimeScript from takara TM RT reagent Kit (Perfect Real Time) and Premix Ex Taq TM (Probe qPCR) kit for reverse transcription and qPCR detection.

[0099] Prepare the reverse transcription reaction system according to the following components (the reaction solution is prepared on ice), and then put it into a PCR machine for reaction. The reaction conditions are 37°C for 60min, 85°C for 5s, and 12°C∞. After the reverse transcription is completed, add 50ul DEPC-H 2 O dilution, take 3ul as template, carry out PCR reaction. The reverse transcription reaction system is shown in Table 3 below.

[0100] table 3

[0101]

[0102] 2. mRNA and LncRNA qPCR

[0103] Prepare the qPCR reaction system according to the following components (the reaction solution is prepared on ice), and set a no-template control as a...

Embodiment 3

[0108] Example 3 Evaluation of the detection effect of the early diagnosis of lung cancer with the external reference β-Actin (ACTB) as the reference single marker

[0109] 1. Sample collection

[0110] Collect 10ml of blood from control samples such as patients with early (stage I and stage II) pulmonary nodules diagnosed by the hospital, patients with benign pulmonary nodules, and healthy people, and separate into plasma.

[0111] 2. Exosomal RNA extraction

[0112] Plasma exosomes were separated by ultracentrifugation or Exosupur of Echobiotech (Beijing Enze Kangtai), and the isolated exosomes were extracted with Qiagen miReasy mini kit kit to extract long RNA in exosomes, and the RNA concentration and quality were detected by Agilent 2100 , and record the RNA concentration.

[0113] 3. RNA two-step detection system

[0114] Using the two-step detection system based on PCR platform mRNA in Example 1, the plasma exosomal mRNA and LncRNA of 30 cases of early lung cancer pa...

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Abstract

The invention discloses a kit, a device and a method for lung cancer diagnosis. The kit comprises exosome long RNA markers, including one or more of LRRC37A2, TLE4, AZI2, MRPL9, YPEL2, RAB3GAP1, TBC1D23, STK3, IL1B, HMGN1, ERI3, HOPX, CDA, CCBL2, TBK1, RAD54L2, ARPC5, GIMAP5, PLA2G1B, SNHG5 and RPL21. The lung cancer detection method based on the exosome long RNA markers has high sensitivity and high specificity in early lung cancer, and provides an important value for early diagnosis of lung cancer, thereby providing a great help to the prevention and treatment of lung cancer in China.

Description

technical field [0001] The invention relates to the field of medical diagnosis, in particular to a diagnostic kit, device and method for early lung cancer. Background technique [0002] Lung cancer is one of the main cancers in my country and even in the world. According to the latest global cancer statistics in 2018, lung cancer ranks first in the incidence and mortality of all cancers. According to the Chinese tumor registration data in 2019, there were 3.902 million new cancer cases in China in 2015, and about 2.338 million cancer deaths. Among them, lung cancer is the main cause of cancer death in China. Therefore, in order to improve the diagnosis and treatment behavior of lung cancer in my country and improve the prognosis of lung cancer patients, early diagnosis has become an important issue in the diagnosis and treatment of lung cancer. [0003] With the application of low-dose spiral CT, more and more imaging manifestations of pulmonary nodules (single lesions <...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/166C12Q2600/178Y02A50/30
Inventor 赫捷高树庚高亦博郭威
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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