Method for detecting lung cancer relative CYP2A13 resistance gene and its resistance gene
A CYP2A13C3375T, resistance technology, applied in the field of individual resistance and its genetics detection, can solve problems such as errors, no literature reports, non-specific amplification analysis results, etc.
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Embodiment 1
[0032] In this example, a set of primer pairs is designed and synthesized, and based on the primer pairs, a method for detecting the functional polymorphism of the resistance allele CYP2A13 C3375T associated with the risk of lung cancer is designed.
[0033] 1. Extraction of genomic DNA:
[0034] The genomic DNA of peripheral blood leukocytes was extracted by Miller's modified salting-out method conventional in the art.
[0035] 1) Take sodium citrate or EDTA-Na 2 5ml of anticoagulated whole blood (try not to use heparin for anticoagulation), put it in a 50ml centrifuge tube with a lid;
[0036] 2) Add 3-5 times the volume of cold distilled water, invert and mix repeatedly, place in ice for 5 minutes, and centrifuge at 2 000 rpm at 4°C for 20 minutes;
[0037] 3) Slowly pour the supernatant, add pre-cooled 0.1% Triton-X100 (same volume as above) to the precipitation, mix the precipitation gently, centrifuge as above, and discard the supernatant;
[0038] 4) Add 4 ml of lysi...
Embodiment 2
[0049] This example uses the method established in Example 1 to analyze the distribution of CYP2A13 C3375T genotypes in 724 lung cancer patients and 791 healthy controls.
[0050] 1. Test subjects: The 724 patients and 791 controls targeted in this example are all individuals of the Han nationality in northern China, with no direct kinship. The case group were all patients with primary lung cancer diagnosed by histopathology who were admitted to the Cancer Hospital of the Chinese Academy of Medical Sciences from 1997 to 2001, and had not received radiation and anticancer chemotherapy before surgery. Normal controls without tumor history and clinical signs were obtained from a large-scale nutritional epidemiological survey conducted in the Beijing area. The sex and age of the control group were matched with the frequency of the case group. While collecting tissue samples or blood samples, each research subject signed an informed consent and provided detailed sociodemographic in...
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