Method and reagent kit for determining full length sequence of ABO gene based on NGS technology

Abstract

The invention belongs to the technical field of genotype detection methods, and in particular relates to a method for determining a full-length sequence of an ABO gene based on NGS technology. The invention also relates to reagents used in the method for determining the full-length sequence of the ABO gene based on the NGS technology. According to the method provided by the invention, the full-length sequence of the ABO gene includes a 5'UTR partial region, and sequences from exon 1 to exon 7 and containing all introns and is subjected to long fragment PCR amplification, amplified products issubjected to library preparation, Illumina platform sequencing on a computer is carried out, and bioinformatics analysis is carried out. The reagents and method provided by the invention can be used as a brand-new, independent and widely-used identification method, the problem of ABO accurate typing is solved, the characteristics of high throughput and accurate results of the NGS technology for ABO genotyping are given full play, and relevant applications in the fields such as clinical blood transfusion medical research and genetics will be highly valued, so that the reagents and method provided by the invention have important practical significance for medical research units, and pharmaceutical research and reagent development units.

Description

technical field [0001] The invention belongs to the technical field of genotyping detection methods, in particular to a method for determining the full-length sequence of an ABO gene based on NGS technology. The invention also relates to a reagent package for determining the full-length sequence of the ABO gene based on NGS technology. Background technique [0002] The ABO blood group system is the most clinically significant blood group system among the 39 known erythrocyte blood group systems. Accurate typing and diagnosis of the ABO blood group antigen system can clarify individual blood group genotypes. [0003] The ABO blood group system gene is located on chromosome 9, the full length of the genome is 19514b, (the 5'UTR and 3'UTR regions are not calculated), and it consists of 7 exons and 6 introns. At present, the genotyping techniques of ABO blood group system include PCR-SSP, PCR-SSO, PCR-SBT, real-time fluorescent PCR technology, gene chip and so on. Except for t...

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Problems solved by technology

Except for the PCR-SBT method among the above methods, several other methods are designed based on known polymorphic sites, and the range of base sequences determined is limited; although the PCR-SBT method overcomes the above-mentioned defects, this method is only limited It is to detect the sequence of the coding region, and the operation process is cumbersome, so it is difficult to achieve high-throughput detection

Studies have found that the mutation points of ABO subtype specimens or their formation mechanisms involve a wide range of sequences. It is necessary to cover the full-length sequence of ABO to identify the subtype and clarify the possible formation mechanism. However, the existing molecular diagnostic methods are difficult to meet the requirements of this category. need

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