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

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Problems solved by technology

However, as mentioned above, tuberculosis, lung adenocarcinoma, and pulmonary sarcoidosis are not easy to distinguis

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

Example Embodiment

[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 pulmonary tuberculosis patients (TB), lung adenocarcinoma patients (AD) and pulmonary sarcoidosis (SA) patients to be tested, and use the lung adenocarcinoma patients' Paracancerous normal tissue served as a normal lung tissue control (NC; n=40).

[0047] 2) Lung tissue samples were ground, mRNA was extracted and purified using RNeasy Plus Mini Kit (Qiagen), and then used III1st Strand cDNA Synthesis SuperMix (Yeasen) kit was reverse transcribed to generate cDNA.

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

Example Embodiment

[0056] The preparation and use of embodiment 2 detection kit

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

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

[0059] The method of using the kit is as follows: taking a lung tissue sample from the subject, extracting RNA from the lung tissue sample using conventional methods (or using a specific kit), using the reagents in the detection kit, and using the normal lung tissue sample in the kit The cDNA sequence of SEQ ID NO. 1 to 6 was used as the control cDNA in the quantitative detection of Real-Time PCR, and the relative expression changes of at least one gene in the lung tissue of the subjects were detected.

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

Example Embodiment

[0061] The application of embodiment 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, chest computed tomography) scan, etc.) to compare the results. The results show that the diagnostic results obtained by using the diagnostic markers of the present application are 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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