Kit and method for typing HLA gene

Abstract

The invention provides a kit and method for typing an HLA gene. The kit comprises a primer for amplifying a second exon and a third exon of an HLA-B gene and a primer for amplifying a fourth exon of the HLA-B gene, wherein the primer for amplifying the second exon and the third exon of the HLA-B gene comprises an amplification primer pair composed of primer sequences as shown in SEQ ID NO.1 and SEQ ID NO.2; and the primer for amplifying the fourth exon of the HLA-B gene comprises an amplification primer pair composed of primer sequences as shown in SEQ ID NO.3 and SEQ ID NO.4. By using the kit and method provided by the invention, the typing process can be simpler, the typing resolution ratio and accuracy rate are increased while the cost is reduced, and furthermore the kit and method are further applied to scientific research and clinic work.

Description

technical field [0001] The invention relates to a kit and a method, in particular to a kit and a method for HLA genotyping. Background technique [0002] There is a method for HLA genotyping by PCR amplification combined with Sanger sequencing. Taking HLA-B genotyping as an example, the implementation plan of this method: first, design specific primers to amplify the No. 2 and No. 3 exons of the HLA-B gene. In order to avoid the bias of the primers, generally design multiple pairs of primers . Secondly, the PCR product was sequenced by Sanger sequencing technology, and the sequencing result containing biallelic information was obtained. Furthermore, the data is analyzed through the company's special software (technical parameters are confidential), so as to obtain the HLA-B typing results of the samples, with a resolution ranging from 4 to 6 digits. [0003] The shortcoming of above-mentioned technique has the following points (taking HLA-B genotyping as example): 1, PCR ...

AI Technical Summary

Problems solved by technology

[0003] The shortcoming of above-mentioned technique has the following points (taking HLA-B genotyping as example): 1, PCR specific primer aspect: existing primer design is more numerous and diverse, needs many pairs of primers to amplify a single exon, and the cost is high , increasing the complexity of the experiment

2. In terms of typing accuracy: only rely on exon 2 and exon 3 sequence information for typing, and the resolution of typing results is low

3. Data analysis: The technical parameters of the company's typing software are kept secret, and the strategies it adopts cannot be evaluated and optimized

[0004] Reasons for the above shortcomings: 1. Design multiple pairs of primers to amplify the target fragment by PCR, because the designer did not make full use of the information in the snp database on the 1000genome and UCSC website to design primers for unbiased amplification

2. Only relying on exon 2 and exon 3 sequence information for typing is because the designer did not consider carefully and ignored that other exons of the HLA-B gene will also affect the typing results

3. The technical parameters of data analysis are kept confidential because the company needs to use the software for profit-making operations. We cannot obtain technical parameters, so we have no way to evaluate them.

Images