Primer, probe, kit and method for detection of human ALDH2 gene polymorphism

Abstract

The invention belongs to the technical field of gene engineering and discloses a primer and a probe combination for detection of human ALDH2 gene polymorphism, a kit comprising the primer and the probe combination, and a fluorescent PCR method for performing the detection of human ALDH2 gene polymorphism with the primer and the probe combination. The primer, the probe and the kit are based on TaqMan fluorescent PCR technology, and are simple, quick and high-sensitive. In addition, through reasonable combination of primer and probe, interaction between primer and primer, primer and probe and probe and probe can be avoided effectively, thus reducing detection error. The primer, the probe and the kit, when being used for the detection of human ALDH2 gene polymorphism, have high sensitivity and strong specificity, are easy, quick and safe to operate, have simple and direct results, and can be applied directly with a blood sample or a dried blood spot specimen on filter paper as a template.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to primers, probes, kits and methods for detecting human ALDH2 gene polymorphisms. Background technique [0002] Nitroglycerin is an emergency drug for the prevention and treatment of coronary heart disease and angina. The enzyme promotes calcium ions to enter the sarcoplasmic reticulum and the extracellular space, thereby dilating vascular smooth muscle and relieving the symptoms of cardiac ischemia. The polymorphism of ALDH2*2 (Glu504Lys, rs671) leads to the substitution of glutamic acid at position 504 of the encoded protein by lysine, and the ALDH2 enzyme activity of individuals carrying the mutant allele ALDH2*2 decreases, and the enzyme activity of heterozygotes is only wild type In 10% of individuals, the mutation is homozygous for loss of enzyme activity. Therefore, patients with ALDH2*2 type cannot use nitroglycerin to resist myocardial ischemia. The carrier ...

AI Technical Summary

Problems solved by technology

Although the sequencing method is the "gold standard" for gene polymorphism detection, it requires a long detection time, complicated operation, and low sensitivity; the PCR-gene chip method has high operational requirements, and hybridization and washing must be protected from light operation, and the chip is expensive; the PCR-melting curve method is non-specific and cannot precisely locate the position of the gene change, and the difference in Tm caused by a single base difference is not very obvious, which usually results in inconspicuous peaks on the melting curve Therefore, there are certain difficulties in the clinical promotion of these three methods

The traditional fluorescent quantitative PCR method is widely used clinically. This method has certain requirements on the number of samples and the distribution of polymorphisms. When the sample size is small, it cannot accurately detect the polymorphisms of the samples. It cannot meet the requirements of high efficiency, accuracy, simplicity, etc. At the same time, it also needs to perform cumbersome and complicated operation procedures such as genomic DNA extraction on the sample, so there is an urgent need for a fast, effective, accurate detection method that does not require DNA extraction

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