Primer composition for guiding nitroglycerin medication and healthy drinking, multiple gene detection kit and use method of kit

Abstract

The invention discloses a primer composition for guiding nitroglycerin medication and healthy drinking, a multiple gene detection kit and a use method of the kit, and the kit comprises the primer composition, a PCR buffer solution and a positive reference substance, and the PCR buffer solution comprises ultrapure water, an X solution, a 10*PCR (polymerase chain reaction) buffer solution, a PCR primer, a 25mM magnesium chloride solution and DNA polymerase, the primer composition comprise two forward and reverse amplification primers of different gene types on the 2 SNP sites of genes related to the nitroglycerin medication and healthy drinking and forward and reverse amplification primers capable of reflecting internal reference, the gene sequences of the primers are represented as SEQ ID NO.1-NO.8; the use method comprises the step of acquiring a sample and extracting nucleic acid, the step of performing the PCR reaction by using extracted nucleic acid as a template, and the final step of separating the sample through capillary electrophoresis. The primer composition has the advantages of being strong in specificity, high in accuracy, high in flux, strong in reliability, low in cost and free from 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 nitroglycerin medication and healthy drinking, a multiple gene detection kit and a use method thereof. Background technique [0002] Nitroglycerin is a nitro vasodilator, and it is one of the special and commonly used drugs for preventing and treating angina pectoris of coronary heart disease. Nitroglycerin and other nitro-containing drugs (such as sodium nitroprusside) can generate active NO free radicals in the body, thereby activating guanylate cyclase in smooth muscle and other tissues, and increasing the synthesis of cGMP. cGMP activates cGMP-dependent protein kinases, alters the phosphorylation of different proteins in smooth muscle, and dephosphorylates the light chain of myosin. Phosphorylation of myosin light chains is known to be involved in maintaining the contractile state of smooth muscle, and experiment...

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

[0015] At present, there are no relevant reports at home and abroad about the multiple SNP detection kit based on multiplex PCR and CE separation to guide the use of thiazide diuretics and its use

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