Immune repertoire method by detecting primers of BCR heavy and light chains in blood and application

Abstract

The invention provides an immune repertoire information analysis method for detecting BCR heavy and light chains in blood. The method specifically comprises the steps of constructing a reference sequence, preprocessing sequencing data, merging Paired Reads, comparing data after Merge with the reference sequence, filtering a comparison result, and performing statistical analysis. Meanwhile, the invention further provides an immune repertoire method for detecting the blood BCR. The information analysis method is adopted in the method, sequencing analysis of a CDR3 region sequence is realized bydesigning a primer combination of the blood BCR heavy and light chains; the method is used for detecting the BCR immune repertoire heavy chain and light chain of rabbits for the first time. CDR3 sequence information related to antibody generation is studied through immune repertoire sequencing of blood B cells, the law of antibody generation is understood more deeply, and meanwhile, a new way forefficiently and rapidly synthesizing specific antibodies in vitro is explored by combining technologies such as gene cloning, cell transfection, protein determination and purification.

Description

Technical field [0001] The invention belongs to the technical direction of immune repertoire sequencing technology in the field of biotechnology, and specifically relates to a BCR heavy chain and light chain special primer set for detecting blood DNA based on multiplex PCR and high-throughput sequencing and a method for constructing an immune repertoire library. A method and application of primers and immune repertoires for detecting blood BCR heavy chain and light chain. Background technique [0002] Immune cells in the human body mainly include T cells and B cells. The T cell antigen receptor is a molecular structure that specifically recognizes and binds antigen peptide-MHC molecules. Most T cell receptors are composed of two peptide chains, α and β, and a few T cell receptors are composed of two peptide chains, γ and δ. Among them, the variable regions (V regions) of the two peptide chains of α and β undergo gene rearrangement during the maturation of immune cells, thereby f...

AI Technical Summary

Problems solved by technology

This method can ensure the specificity of the amplification product region, but if the number of amplification primers is too large, the amplification product will be inhomogeneous

(2) The generally designed primer combination is a V region subtype corresponding to a set of primer pairs. If multiple V region subtypes are included, the complexity of the primer combination will increase, and it is easy to cause different primer pairs to amplify during multiplex PCR amplification. There are differences in the efficiency of the increase, which in turn affects the accuracy of the subsequent analysis of the sequence changes in the CDR3 region

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