H63D detection classification reagent kit based on AllGlo probe and classification method thereof

Abstract

The invention discloses an H63D detection classification reagent kit based on an AllGlo probe and a classification method thereof, and relates to single nucleotide polymorphism. The reagent kit comprises a real-time fluorescence quantification PCR reagent, a positive control and a negative control. H63D is an SNP locus in an HFE gene of the human chromosome 6 provided by NCBI. The detection classification method comprises the steps that 1, a conventional method is adopted for extracting DNA in an EDTA anti-coagulation whole-blood sample; 2, the real-time fluorescence quantification PCR reagent provided by the H63D detection classification reagent kit based on the AllGlo probe is adopted for conducting real-time fluorescence quantification PCR amplification on the DNA; 3, according to detected fluorescence signals, human HFE gene single nucleotide polymorphism sites H63D are classified. On the premise of maintaining the high specificity and sensitivity, the detection price of the AllGlo probe is lower than that of a direct sequencing method, the process is easier, and consumed time is shorter.

Description

technical field [0001] The present invention relates to a single nucleotide polymorphism (SNP), in particular to an AllGlo probe-based H63D detection and typing kit and typing method thereof. Background technique [0002] Single nucleotide polymorphism (singlenucleotide polymorphism, SNP) mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genome level. It is the most common type of heritable variation in humans. SNPs widely exist in the human genome, with an average of 1 in every 500-1000 base pairs, and it is estimated that the total number can reach 3 million or more. As a third-generation genetic marker, SNP is closely related to many phenotypic differences, drug susceptibility and disease susceptibility. The large number of SNPs in the genome makes it a powerful tool and plays a very important role in disease localization and cloning, drug design and testing, and basic biological research. [0003] Individualized diagn...

AI Technical Summary

Problems solved by technology

Among them, the direct sequencing method is currently the most intuitive and relatively accurate SNP typing method, but its steps are many and scattered, the cost is high, the workload is heavy, the cycle is long, and the price is expensive, so it is not suitable for large samples and multiple loci. Detection; Restriction Fragment Length Polymorphism (RFLP), as the earliest DNA labeling technology, has low requirements for instruments, and only needs a PCR instrument and an electrophoresis instrument to carry out the experiment, but its experimental operation is cumbersome and the detection cycle is long , not suitable for large sample detection; Tagman fluorescent probe method has good accuracy, but its design and synthesis procedures are complicated, and it is not suitable for typing of multiple loci and a small number of samples, especially loci with low frequencies; amplification block mutation The system (ARMS method) is fast and simple, but it cannot be operated in a closed tube, and it is difficult to achieve high-throughput and automation for gene polymorphism analysis; high-resolution melting curve analysis (HRM) has the advantages of simplicity and speed, but the method is specific Insufficient, few instrument choices

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