HLA-B*5801 genetype rapid detection kit based on POCT mode

Abstract

The invention provides an HLA-B*5801 genetype rapid detection kit based on a POCT mode. The kit at least contains fluorogenic quantitative PCR primers and probes (SEQ ID NO:1-4) as well as a cell lysis solution, and can further comprise dNTPs, DNA polymerase, Mg<2+>, a reaction buffer, a standard positive template, a sampling bar, a sample collecting tube and the like, wherein the fluorogenic quantitative PCR primers and the probes are used for detecting the HLA-B*5801 (rs9263726) genetypes. The kit can realize instant detection, DNA purification is not needed, a sample can be directly added into a reagent for carrying out a PCR reaction, the kit is particularly suitable for rapidly and accurately detecting samples (such as shedding buccal cells) with low content of DNA, the detection accuracy rate achieves 99% or above, the detection sensitivity is high, and genomic DNA of which the amount is as low as 0.125ng can be accurately detected; and the whole detection consumes short time, adetection result can be obtained within 1h, an administration basis can be provided for a doctor at first time, and thus the administration risk of patients is reduced.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a POCT mode-based rapid detection kit for HLA-B*5801 genotype. Background technique [0002] Human leukocyte antigen HLA is a group of genes located on chromosome 6, which is the most complex genetic polymorphism system in humans, and its polymorphism distribution has obvious ethnic characteristics. [0003] Allopurinol, which inhibits uric acid production, is one of the most common drugs causing skin adverse reactions. According to statistics, about 2% of patients taking allopurinol have skin allergic reactions of varying severity clinically, although less than 1% of the cases will turn into drug hypersensitivity syndrome, Stevens-Johnson syndrome and toxic epidermal necrolysis Severe skin adverse reactions including symptoms, but its mortality rate is as high as 10%-40%. [0004] Studies have found that genetically specific side effects of drugs are often closely related to h...

AI Technical Summary

Problems solved by technology

According to statistics, about 2% of patients taking allopurinol have skin allergic reactions of varying severity clinically, although less than 1% of the cases will turn into drug hypersensitivity syndrome, Stevens-Johnson syndrome and toxic epidermal necrolysis severe skin adverse reactions including symptoms, but its mortality rate is as high as 10%-40%

Although this method has the advantages of low cost, short time and simple result analysis, since the reaction system can only amplify purified genomic DNA samples, multi-step and multi-tube operations must be used, which is not ideal for clinical promotion.

Therefore, these conventional detection techniques are difficult to meet the requirements of fast and accurate HLA-B*5801 gene detection for the population carrying the HLA-B*5801 mutation gene in China

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