One group of primer for detecting SNP (single nucleotide polymorphism) site rs9263726, crRNA (CRISPR (clustered regularly interspaced short palindromic repeat sequences) RNA) sequence and application of primer

Abstract

The invention provides a system for detecting the SNP (single nucleotide polymorphism) site rs9263726 of a HLA-B*5801 gene. The system comprises a primer for detecting the SNP site rs9263726, two (CRISPR (clustered regularly interspaced short palindromic repeat sequences) RNA) sequences, two different CRISPR / cas13 protein systems and reporter probes with different fluorescent marks. When the system disclosed by the invention is used for detecting a mutation type of the SNP site rs9263726 of the HLA-B*5801 gene, the system is convenient in operation, and each detection sample only needs to be carried out in a single tube; a whole reaction process is carried out at a temperature of about 37 DEG C instead of a delicate temperature control element and complex temperature change just like PCR (Polymerase Chain Reaction) amplification, and therefore, the system is suitable for grassroots units to carry out detection by relatively cheap instant detection equipment; and compared with a PCR technology, the system has a higher speed, can obtain an obvious detection signal in 30 min, is high in detection result specificity and can easily judge the type of an allele.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a group of amplification primers for detecting PSORS1C1 gene polymorphic site rs9263726 and application thereof. Background technique [0002] Allopurinol is the first-line drug for the treatment of gout and is currently the only xanthine oxidase inhibitor. The drug can cause serious side effects, including Stevens-John syndrome (SJS) and toxic epidermal necrosis (TEN). The strong correlation between the HLA-B*5801 allele and allopurinol-induced SJS or TEN was first discovered in the Han population by scholars in Taiwan, China, and confirmed by multiple research groups in Thailand, Singapore, Hong Kong, and China in the Han population , the HLA-B*5801 allele frequency in the Han people is 9-15% (geographical differences). Studies have shown that there is a strong correlation between the HLA-B*5801 allele and the severe dermatitis side effects caused by allopu...

AI Technical Summary

Problems solved by technology

Such a method shows a certain specificity in double-tube detection, but in some cases, especially in the presence of heterozygotes, the difference in detecting different alleles (alleles) is still not significant, and it is difficult to intuitively judge the detection of samples, etc. allele type

The second method is mainly used for multiple detection of different genes, but it is difficult to implement at the SNP site (wild type / mutant type)

In addition to the interference of the specific response due to the high homology between the target ssRNAs at different SNP sites, the strength of the side activation response between different Cas13s is also significantly different; in addition, the preference or specificity of the side cutting base sequences of different Cas13 proteins is not Absolutely, it will cut non-specifically and reduce the specificity of the response

The superposition of these aspects makes it difficult for the operator to intuitively judge the specific SNP type of the test product through the signal intensity of different fluorescence

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