Gene marker for identifying benign and malignant cases of lung micro nodules and application of gene marker
A tiny, nodular technology, applied in the field of molecular biology, can solve problems such as false positives, low detection sensitivity, and low diagnostic value, and achieve the effect of promoting early detection, broad application prospects, and non-invasive sampling methods
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Embodiment 1
[0026] Example 1 Screening of peripheral blood characteristic gene markers of lung cancer
[0027] The screening of lung cancer characteristic genes includes the following steps:
[0028] 1) Collect peripheral blood samples from 40 patients with malignant micronodules diagnosed as lung cancer by surgery and pathological examination, 16 patients with benign micronodules (benign lung lesions), and 28 healthy people without lung lesions. All patients with micronodules were confirmed by CT examination that the size of the nodules was less than or equal to 10mm, and 2-3ml of peripheral blood was collected from each sample.
[0029] 2) The PAXgene Blood RNA Kit extraction kit was used to extract the total RNA of the above samples, the fragment integrity (RIN) of the RNA sample was detected by the AgilentBioanalyzer 2100 bioanalyzer, and the purity of the RNA sample was detected by the Nano1000 micro-volume ultraviolet spectrophotometer. All RNA samples must meet the following quality...
Embodiment 2
[0038] Example 2 Using the screened lung cancer characteristic gene markers to detect micronodular lung cancer
[0039] 1. Method steps:
[0040] 1) Collection of peripheral blood samples of samples to be tested: use BD PAXgeneRNA blood collection tubes from QIAGEN to collect peripheral blood samples from patients;
[0041] 2) Extraction and purification of total RNA in the peripheral blood sample of the sample to be tested: use QIAGEN's PAXgeneBloodRNAKit to extract and purify the total RNA in the peripheral blood, and use the Agilent BioAnalyzer 2100 microelectrophoresis analyzer to identify the integrity and yield of the extracted total RNA fragments. Use Nano1000 micro-volume ultraviolet spectrophotometer to detect the purity of RNA samples;
[0042] 3) Reverse transcription reaction: use Life Techonolgy's High-Capacity cDNA ReverseTranscription kit reverse transcription kit, use total RNA as a template, and use Olig(dT) as a reverse transcription primer to perform a reve...
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