Pulmonary nodule diagnosis marker and application

A lung nodule, PLA2G6 technology, applied in the field of biotechnology detection, can solve problems such as difficult to distinguish, and achieve good clinical application prospects

Pending Publication Date: 2020-11-03
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as mentioned above, tuberculosis, lung adenocarcinoma, and pulmonary sarcoidosis are not easy to distinguish in many cases, so effective disease-specific lung tissue biomarkers are needed for auxiliary detection and judgment

Method used

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  • Pulmonary nodule diagnosis marker and application
  • Pulmonary nodule diagnosis marker and application
  • Pulmonary nodule diagnosis marker and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 The mRNA levels of PTGER4, AKR1C1, PLA2G6, LTA4H, PLA2G7 and AKR1C3 were significantly increased in the diseased lung tissue of patients with pulmonary sarcoidosis

[0046] 1) Obtain lung tissue samples (n=35, 48 and 21) from patients with pulmonary tuberculosis (TB), lung adenocarcinoma (AD) and pulmonary sarcoidosis (SA) to be tested, and use lung adenocarcinoma patients Adjacent normal tissue was used as normal lung tissue control (NC; n=40).

[0047] 2) Lung tissue samples were ground, and mRNA was extracted and purified using RNeasy Plus Mini Kit (Qiagen), and then used Ⅲ 1st Strand cDNASynthesis SuperMix (Yeasen) kit was reverse transcribed to generate cDNA.

[0048] 3) Using Applied Biosystems 7500 Real-Time PCR system, perform qPCR to analyze the above cDNA samples, and detect the mRNA levels of PLA2G6, PLA2G7, AKR1C1, AKR1C3, LTA4H and PTGER4. ACTB and 18S rRNA were used as internal reference genes. The primers used are as follows:

[0049] Table...

Embodiment 2

[0056] Preparation and use of embodiment 2 detection kit

[0057] The nucleotide sequences shown in Table 1 were synthesized as primers for amplifying internal reference genes and markers.

[0058] For convenience, the kit can also contain a control: the cDNA sequence of a normal human lung tissue sample.

[0059] The method of using the kit is as follows: take a lung tissue sample from the subject, extract RNA from the lung tissue sample using a conventional method (or use a specific kit), use the reagents in the detection kit, and use the normal lung tissue sample in the kit The cDNA sequence is used as the control cDNA in the Real-Time PCR quantitative detection, and the relative expression level change of at least one gene in the lung tissue of the subject such as SEQ ID NO.1-6 is detected.

[0060] The expression of PLA2G6 is more than 2.851 times of the control, or the expression of PLA2G7 is more than 3.347 times of the control, or the expression of AKR1C1 is more than...

Embodiment 3

[0061] Application of Example 3 Markers in Clinical Diagnosis

[0062] According to the method of Example 1, multiple subjects were tested respectively, and the test results were compared with current diagnostic methods (such as blood test, sarcoidosis antigen test, biopsy, lung X-ray examination, chest computed tomography) scans, etc.) for comparison. The results showed that the diagnostic results obtained by using the diagnostic markers of the present application were consistent with the pathological diagnostic results.

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Abstract

The invention discloses a pulmonary nodule diagnosis marker and an application, and belongs to the technical field of biotechnology detection. The invention provides an application of one or more of PLA2G6, PLA2G7, AKR1C1, AKR1C3, LTA4H and PTGER4 genes as biomarker molecules for diagnosing pulmonary nodule diseases, and the biomarkers can be used for assisting in living tissue examination and helping to confirm related pulmonary diseases. The accuracy rates (areas under ROC curves) of the genes for identifying the sarcoidosis in distinguishing healthy people, lung adenocarcinoma patients andpulmonary tuberculosis patients are 96.5%, 90.9%, 89.1%, 87.5%, 87.2% and 85.8% in sequence, and the comprehensive accuracy rate is 90.9%. According to the invention, a certain quantity of clinical samples are collected for research and verification, the research result conforms to clinical practice, and the pulmonary nodule diagnosis marker has good clinical application prospects.

Description

technical field [0001] The invention relates to pulmonary nodule diagnostic markers and their application, in particular to the development of products such as a kit for diagnosing pulmonary sarcoidosis using patient lung tissue samples, and belongs to the technical field of biotechnology detection. Background technique [0002] Pulmonary nodule (sarcoidosis) is a multi-system multi-organ granulomatous disease of unknown etiology, which often invades the lungs, bilateral hilar lymph nodes, eyes, skin and other organs, and the chest invasion rate is as high as 80% to 90%. Clinically, pulmonary nodules have very similar symptoms, pathological features, and imaging findings to many other chronic lung diseases, such as lung cancer and tuberculosis, which pose a great challenge for differential diagnosis. Lung cancer is the type of cancer that causes the most human deaths. In 2018, the number of deaths due to lung cancer in the world exceeded 1.7 million. Among them, lung adenoc...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/158C12Q2600/166G01N33/6884G01N2800/12G01N2800/52G01N2800/60
Inventor 刘翠华柴琪瑶陆喆汪静
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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