A primer set and kit for detecting genes related to hypertension drug metabolism
A primer set and high blood pressure technology, applied in the fields of medicine and biology, can solve the problems of cumbersome operation, long detection cycle, uncontrollable factors of test results, etc., achieve simple sample processing, high sensitivity and specificity, and reduce adverse drug reactions Effect
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Embodiment 1
[0097] Embodiment 1: the preparation of kit
[0098] 1. Design and synthesis of primers for genes related to hypertension drug metabolism
[0099] Using oligo software to design BDKRB2 gene, CYP11B2 gene, OAT1 / OAT3 gene, AGT gene, KCNH2 gene, LDLR gene, APOB gene, CACNA1C gene, GRK4 gene, ACE gene, CYP2D6 gene, CYP2C9 gene, CYP2C19 gene, ACE2 gene, AGTR1 Gene, ADRB2 gene and NR3C2 gene, the PCR amplification primers and sequencing primers of 17 hypertensive drug metabolism-related genes (see Table 1). The primer design follows the following principles: the length of the primer is about 25-30bp; the primer has no dimer; no hairpin structure; the Tm value difference between the upstream and downstream primers should not exceed 3°C; the GC content is about 50%. After the upstream and downstream primers are determined, use Blast to compare and analyze the upstream and downstream primers in the NCBI database to ensure the specificity and amplification efficiency of the primers; ...
Embodiment 2
[0106] Embodiment 2: the use of kit
[0107] 1. Sample testing
[0108] Take out the relevant reagents, and prepare the corresponding system according to the amount of template as follows: Take the PCR reaction solution, mix it evenly, add sample DNA, negative control substance or positive control substance as template to form a reaction system. Perform PCR amplification according to the PCR reaction procedure.
[0109] The main components of each gene detection system for hypertension are shown in Table 3.
[0110] Table 3. Main components of each gene detection system for hypertension
[0111]
[0112] The PCR reaction program of this system is shown in Table 4.
[0113] Table 4. PCR reaction program
[0114]
[0115] After the amplification is completed, agarose gel electrophoresis is performed to check the PCR amplification product, and the PCR amplification product is purified before proceeding to the next step.
[0116] 2. Sanger sequencing
[0117] Follow ...
Embodiment 3
[0133] Example 3: The effect of the primer set and / or kit of the present invention on the medication guidance of hypertensive patients
[0134] In order to verify the effect of the primer set and / or kit of the present invention on the drug guidance effect of hypertensive patients, volunteers with hypertension were selected at the "Present Medical Laboratory", and the genotype was performed using the invented primer set and / or kit. Test, according to the test results of each volunteer, accurately formulate its medication plan, and conduct clinical follow-up to observe the clinical drug effect.
[0135] Medication Guidance Example 1
[0136] The tester Tian, who took spironolactone before the test, did not have a good effect. Using the primer set and / or kit of the present invention, Tian was tested for genes related to hypertension. According to Tian's gene detection results, he predicted hydrochlorothiazide and Propranolol is more effective. After Tian took propranolol ...
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