Method for detecting genotype of gene CYP1A2 associated with medicament metabolism ability and liver cancer susceptibility
A metabolizing ability and related technology, applied in the field of detecting the gene CYP1A2 genotype associated with drug metabolizing ability and liver cancer susceptibility, can solve the problem that individual differences in CYP1A2 activity cannot be fully explained.
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Embodiment 1
[0042] In this example, a set of primer pairs was designed and synthesized, and based on this primer pair, a method for detecting three SNPs of genes CYP1A2G-3860A, G-3113A and T5347C associated with drug metabolism ability and susceptibility to liver cancer was designed locus genotype method.
[0043] 1. Genomic DNA extraction
[0044] Genomic DNA of peripheral blood leukocytes was extracted by Miller's modified salting-out method, which is conventional in the field.
[0045] 1) Take sodium citrate or EDTA-Na 2 5ml of anticoagulated whole blood (try not to use heparin for anticoagulation), put in a 50ml capped centrifuge tube;
[0046] 2) Add 3-5 times the volume of cold distilled water, invert and mix repeatedly, place in ice for 5 minutes, and centrifuge at 2000rpm at 4°C for 20 minutes;
[0047]3) Slowly pour the supernatant, add ice-precooled 0.1% Triton-X100 (same volume as above) to the precipitate, gently mix the precipitate, centrifuge as above, and discard the sup...
Embodiment 2
[0059] This embodiment uses the method established in Example 1 to genotype the CYP1A2G-3860A, G-3113A and T5347C loci in 419 healthy Chinese population, and observe the genotypes of the three loci. Relationship between plasma caffeine metabolic ratio or CYP1A2 activity in vivo.
[0060] 1. Test object:
[0061] 419 healthy subjects (237 males, 182 females) aged 18-33 (average age 20±2 years old) participated in this experiment. All the subjects are students of Central South University. All subjects were non-smoking individuals, and had not consumed coffee, tea, Coca-Cola, chocolate or other caffeinated beverages within 1 week before the detection of caffeine metabolic rate. At the same time, all the subjects did not take any drugs, including contraceptives, within 1 week before the test. The entire experimental process was carried out in accordance with the National Human Genome Research Ethics Guidelines, and informed consent was obtained from all subjects.
[0062] 2. D...
Embodiment 3
[0070] This embodiment applies the method established in Example 1, and the genotypes and genotypes of CYP1A2G-3860A, G-3113A and T5347C loci of 431 cases of liver cancer patients and 550 cases of control DNA samples from Fusui County and its surrounding counties and cities in Guangxi Zhuang Autonomous Region were analyzed. Haplotype combined genotypes were analyzed.
[0071] 1. Test object:
[0072]From June 2003 to December 2004, we collected a total of 431 liver cancer specimens from Fusui County Liver Cancer Institute and Guangxi Zhuang Autonomous Region Cancer Hospital (about 40 kilometers away from Fusui County). Most of the cases in the Cancer Hospital of Guangxi Zhuang Autonomous Region come from Fusui County and surrounding counties and cities. The diagnosis basis of liver cancer cases included: 89 cases (20.6%) were confirmed by histopathology, 247 cases (57.3%) were positive in imaging diagnosis (B-ultrasound examination and / or CT scan) accompanied by elevated seru...
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