ADRB1 genotype rapid detection kit based on POCT mode

Abstract

The invention provides an ADRB1 genotype rapid detection kit based on a POCT mode. The kit comprises fluorogenic quantitative PCR primers and probes as well as a cell lysis solution, wherein the fluorogenic quantitative PCR primers and the probes are used for detecting the ADRB1 genotypes (G1165C), the sequences of the PCR primers are as shown in SEQ ID NO.1-2, and the sequences of the probes areas shown in SEQ ID NO.3-4. The kit can realize instant detection, DNA purification is not needed, a sample can be directly added into a reagent for carrying out a PCR reaction, the kit is particularlysuitable for rapidly and accurately detecting samples (such as shedding buccal cells) with low content of DNA, the detection accuracy rate achieves 99% or above, the detection sensitivity is high, and genomic DNA of which the amount is as low as 0.1ng can be accurately detected; and the whole detection consumes short time, a detection result can be obtained within 1h, an administration basis canbe provided for a doctor at first time, and thus the administration risk of patients is reduced.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a POCT mode-based rapid detection kit for ADRB1 genotype. Background technique [0002] The β1-adrenergic receptor (ADRB1) belongs to the adrenergic receptor family, is coupled to guanosine nucleoside protein (G protein) and is widely distributed in cardiomyocytes. ADRB1 is a protein receptor containing 477 amino acid residues. After binding to catecholamine, it can activate adenylate cyclase (AC) to generate a large amount of cyclic adenosine monophosphate (cAMP), thereby further activating protein kinase A to regulate heart rate. and blood pressure. [0003] ADRB1 has a single nucleotide polymorphism (SNP), in which the G1165C (rs1801253) site mutation causes the 389th glycine (Gly) to be replaced by arginine (Arg). G1165C is a mutation site significantly associated with hypertension. Therefore, in order to achieve the effect of lowering blood pressure, the activity of ADRB1...

AI Technical Summary

Problems solved by technology

Sequencing method (such as patent CN105586406A) and gene chip method (such as patent CN102021238A) have high precision to the detection results, but need large-scale precision instruments and equipment, fixed laboratories and professional operators, therefore, the cost of these two detection methods is high , the steps are cumbersome and time-consuming, and it is difficult to promote clinically

Although the fluorescent PCR method based on Taqman probes (such as patent CN106834466A) has the advantages of low cost, short time and simple result analysis, because this reaction system can only amplify the purified genomic DNA sample, it must adopt multiple steps and multiple It is not ideal for clinical promotion

Therefore, these conventional detection techniques are difficult to meet the rapid and accurate detection requirements of ADRB1 (G1165C) mutant gene in China.

Images