Method for correcting and standardizing TCR beta high-throughput sequencing data based on template sequence and reference cell

Abstract

The invention belongs to the technical field of biology, and particularly discloses a TCR beta high-throughput sequencing data correction and standardization method based on a template sequence and a reference cell, which comprises the following steps: doping a fixed number of external reference cells and a fixed number of synthetic templates into a sample to construct a TCR beta high-throughput sequencing library, a high-throughput sequencing platform is used for sequencing; analyzing an amplification bias rule by using the added template sequence; a sequencing error caused by base mutation bias is corrected; standardizing the sample sequencing data by using the external reference cells; the T cell receptors in the sample are accurately quantified. According to the method, TCR beta high-throughput sequencing data can be corrected and standardized through amplification bias correction, sequencing error correction and sample standardization in a high-throughput sequencing mode by adopting a standardized process, and finally, accurate and real TCR beta library distribution is obtained.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for correcting and normalizing TCRβ high-throughput sequencing data based on a template sequence and a reference cell. Background technique [0002] T cell receptors (TCRs) are specific receptors on the surface of T cells, responsible for recognizing antigens presented by the major histocompatibility complex (MHC) and mediating immune responses. Understanding the diversity of the T cell receptor repertoire will help us understand the immune status of the body, and then clarify the internal motivation of the occurrence and development of immune diseases, and provide help for the development of related vaccines and the treatment of diseases. Complementarities determining region 3 (CDR3) on the T cell receptor β subunit is a very important region on the TCR receptor. This region has the strongest binding ability to antigenic peptides and is also the region with the highest dive...

AI Technical Summary

Problems solved by technology

However, during the deep sequencing of the T cell receptor repertoire, sequencing errors seriously affect the estimation of the diversity of the T cell repertoire

In addition, in the process of building a library, multiple PCR amplifications are required for the CDR3 sequences in the samples. The interference between multiple primers and the difference in amplification efficiency will cause amplification bias.

Therefore, it is necessary to establish an effective method to correct amplification bias, PCR and sequencing errors, so as to promote the research of T cell receptor library.

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