Primer composition and multiple-gene detection kit for guiding administration of thiazine diuresis drugs and application method thereof

Abstract

The invention discloses a primer composition and multiple-gene detection kit for guiding the administration of thiazine diuresis drugs and an application method thereof. The primer composition comprises ultrapure water, an X solution, a 10*PCR buffer solution, a PCR primer, a 25mM magnesium chloride solution, a DNA polymerase and a positive reference substance and is characterized in that the PCR primer comprises the following ten forward / reverse amplification primers and reaction reference forward / reverse amplification primers of different gene types on 15 SNP sites on the gene related to the administration of thiazine diuresis drugs, wherein the gene sequences are shown by the SEQ ID No.1 to No.46. The application method comprises the following steps: collecting the samples and extracting nucleic acid; performing a PCR reaction by taking the extracted nucleic acid as a template; and finally, performing capillary electrophoretic separation of the samples with the GeXP genetic analyzer. The primer composition has the advantages of strong specificity, high accuracy, high flux, high reliability, low cost and no false negative result.

Description

technical field [0001] The invention relates to a multiple gene detection kit and a detection method thereof, in particular to a primer composition for guiding the administration of thiazide diuretics, a multiple gene detection kit and a use method thereof. Background technique [0002] Thiazide diuretics are one of the five first-line antihypertensive drugs, which mainly exert their effects through three mechanisms: inhibiting the absorption of sodium chloride in the anterior segment of the distal convoluted tubule and the proximal convoluted tubule, and increasing NaCl in the distal convoluted tubule and collecting duct. + -K + Exchange, promote K + secretion; inhibit the activity of phosphodiesterase, reduce the uptake of fatty acids and mitochondrial oxygen consumption by renal tubules, and inhibit the renal tubular response to Na + , Cl - active reabsorption; due to renal tubules for water, Na + The absorption of the kidney decreases, the pressure in the renal tubul...

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 is no relevant report on the kit and its application method based on multiplex PCR and CE multiplex SNP detection to guide the administration of thiazide diuretics at home and abroad

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