Lung cancer detection marker and diagnostic kit

A technology for lung cancer diagnosis and detection reagents, which is applied in biological tests, measurement devices, material inspection products, etc., to achieve the effect of improving sensitivity and specificity, and easy detection.

Inactive Publication Date: 2018-12-07
南京求臻基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Currently, lung cancer detection methods include chest X-ray examination, chest CT, percutaneous lung biopsy, etc., but

Method used

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  • Lung cancer detection marker and diagnostic kit
  • Lung cancer detection marker and diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Detection of CD171 protein expression by real-time fluorescent PCR

[0024] 1. Extraction of total RNA and total protein by TRIzol method

[0025] Total RNA was extracted from the blood sample using the TRIzol method. After RNA extraction, 1 μL was taken and detected by 1% agarose gel electrophoresis, and the concentration was detected by Nanodrop2000 (Thermo Scientific), and the RNA quality was evaluated. The ratio of 260 / 280 was between 1.8-2.1 RNA between RT reagent Kit (TaKaRa) Reverse Transcription Kit Instructions Reverse transcribed into cDNA, and the synthesized cDNA was stored at -20°C for future use.

[0026] 2. Design of primers

[0027] The gene sequences of CD171 and the internal reference β-Actin were obtained according to the Genbank query, and the amplification primers were designed using the software Primer 5.0.

[0028]

[0029] 3. Detection of CD171 expression by real-time fluorescent quantitative PCR

[0030] (1) Reaction system: se...

Embodiment 2

[0039] Example 2 Analysis of sensitivity and specificity of exosomal CD171 protein

[0040] ROC curve analysis showed that CD171 had an AUC value of 0.8574 when distinguishing the healthy group from the lung cancer patient group, and the sensitivity at the optimal cut-off point was 71%, and the specificity was 66% ( figure 2 ), the results show that exosomal CD171 has good sensitivity and specificity as a potential lung cancer detection marker.

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Abstract

The invention discloses a lung cancer detection marker and a diagnostic kit, application of exosome CD171 protein to preparation of lung cancer diagnostic kits as a detection target spot, and application of a reagent for detecting the expression quantity of exosome CD171 protein to preparation of the lung cancer diagnostic kits. A real-time fluorescent PCR detection reagent for detecting the mRNAexpression quantity of the exosome CD171 protein preferably contains qPCR primers for detecting the exosome CD171 protein, wherein an upstream primer sequence is as shown in SEQ ID NO.1, and a downstream primer sequence is as shown in SEQ ID NO.2. The invention discloses the exosome CD171 protein, which can be used for diagnosing whether a patient suffers from lung cancer by detecting the proteinexpression level of CD171. Proved by experiment, the CD171 protein can effectively distinguish a person of lung cancer from a healthy person. Therefore, the invention provides a new detection target spot for the early diagnosis detection of lung cancer.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a lung cancer detection marker and a diagnostic kit. Background technique [0002] Lung cancer (lung cancer) is currently the malignant tumor with the highest incidence and mortality in the world. In the past 50 years, many countries have reported that the incidence and mortality of lung cancer have increased significantly. Among them, the incidence and mortality of lung cancer in men and women are Ranked first and second in all malignant tumors, respectively. Lung cancer has become a common malignant tumor of the chest that seriously threatens human health. [0003] At present, the preferred treatment for lung cancer is surgery. However, even if radical resection is performed, some patients still cannot achieve a high survival rate. At present, about 75% of lung cancer patients are diagnosed at an advanced stage, and the 5-year survival rate is about 15%. The use of bronchos...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/68G01N33/574C12N15/11
CPCC12Q1/6886C12Q2600/158G01N33/57423G01N33/68G01N2333/70503
Inventor 李曼曼段小红承康平丁蕊张磊王东亮周启明
Owner 南京求臻基因科技有限公司
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