Primer combination for guiding application of beta-receptor blocker, multi-gene detection kit and using method thereof

Abstract

The invention discloses a primer combination for guiding the application of a beta-receptor blocker, a multi-gene detection kit and a using method thereof. The kit disclosed by the invention comprises ultrapure water, an X solution, 10*PCR (Polymerase Chain Reaction) buffer, PCR primers, a 25 mM magnesium chloride solution, DNA (Deoxyribonucleic Acid) polymerase and a positive reference substance. The kit is characterized in that the primers include three forward and reverse amplification primers which are different in genotype and located on three SNPS loci on a gene related to the application of the beta-receptor blocker and forward and reverse amplification primers for the internal reference of reaction, and the gene sequences of the primers are shown in SEQ ID NO.1-NO.11. The using method comprises the following steps: collecting samples and extracting a nucleic acid; carrying out a PCR reaction by taking the extracted nucleic acid as a template; finally, carrying out capillary ionophortic separation on the samples by using a GeXP genetic analyzer; the using method has the advantages of strong specificity, high accuracy, high flux, high reliability, low cost, no false negative results.

Description

technical field [0001] The invention relates to a multiple gene detection kit and a use method thereof, in particular to a primer composition for guiding the administration of beta-receptor blocking drugs, a multiple gene detection kit and a use method thereof. Background technique [0002] β-receptor blockers are one of the five first-line antihypertensive drugs. Receptor blockers can selectively bind to β-adrenergic receptors, thereby antagonizing the stimulatory effect of neurotransmitters and catecholamines on β-receptors A drug type. Adrenergic receptors are distributed on the effector cell membranes dominated by most sympathetic postganglionic fibers, and the receptors are divided into three types, namely β1 receptors, β2 receptors and β3 receptors. β1 receptors are mainly distributed in the myocardium, which can be excited to increase heart rate and myocardial contractility; β2 receptors exist in bronchial and vascular smooth muscles, and can be excited to cause bron...

AI Technical Summary

Problems solved by technology

Its disadvantages are (1) low throughput: it is not suitable for the detection of multiple SNP sites; it is difficult to set internal control genes

(2) High cost: the cost of probe labeling is high; if you need to obtain all relevant SNP information, you need to conduct multiple detection tests, and the superimposed cost is more expensive

Disadvantages: 1) The hybridization kinetics between different SNP sites are different, and the conditions are difficult to control when performing simultaneous detection of multiple sites; 2) The technology is expensive and complicated: each sample requires a chip, and the cost is more than ￥1000 / sample , which is not conducive to large-scale promotion; the synthesis and fixation of probes are relatively complicated, especially the production of high-density probe arrays, which is the main speed-limiting step; 3) poor repeatability, low accuracy, and false positive and false negative results are prone to occur; 4) Low sensitivity: the chip method requires a large amount of nucleic acid, and generally requires multiple PCR amplification first. Due to the large number of primers, it is easy to produce dimers and hairpin structures by itself, or the purpose of amplification is caused by different Tm values. Different fragment efficiencies will affect the detection sensitivity; 5) Due to the variety of chips, it is difficult to formulate a unified quality control standard

[0014] At present, there are no relevant reports on the kit and its usage method based on multiplex PCR and CE multiplex SNP detection to guide the administration of β-receptor blocking drugs at home and abroad

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