Method for accurately detecting T cell immune repertoire based on high-throughput sequencing and primer system thereof

Abstract

The invention provides a method for accurately detecting a T cell immune repertoire based on high-throughput sequencing and a primer system thereof, the primer system comprises a PCR1 primer and a PCR2 primer, the PCR1 primer sequence and the PCR2 primer sequence comprise an upstream V primer and a downstream J primer, and a sequencing linker sequence and a molecular tag sequence are respectively inserted into target areas of the upstream primer and the downstream primer of the PCR1; the PCR2 primer takes a PCR1 product as a template, and a sequencing primer sequence and an index tag are introduced. The primer system and the method for accurately detecting a T cell immune repertoire based on high-throughput sequencing have the characteristics of simplicity and convenience in sample treatment, rapidness in experimental operation and accurate quantification of TCR cloning clusters.

Description

technical field [0001] The invention relates to the field of molecular biology detection, in particular to a method for accurately detecting T cell immune repertoire based on high-throughput sequencing and a primer system thereof. Background technique [0002] T cell receptor (T cell receptor, TCR) is a molecule on the surface of T cells that specifically recognizes antigens and mediates immune responses. It is one of the most polymorphic regions in the human genome and determines how the human immune system adapts to the environment. Variety. The diversity of T cell receptor library (including gene recombination and selective expression) directly reflects the state of the body's immune response. During the development of T cells, CDR1, 2 and FR regions are relatively conserved, and the CDR3 region is rearranged by V, D and J to form a functional TCR coding gene (T cell clone), because V (65-100 species), D (2 species) and J (13 species) gene fragments themselves have dive...

AI Technical Summary

Problems solved by technology

This method can accurately quantify and detect the TCR immune repertoire, but due to the complexity of RNA sample processing and the fluctuation of RNA expression, its repeatability and specificity are not as good as DNA detection results, so it is not an ideal raw material

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