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
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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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