Method for detecting BCR and TCR immune repertoire in blood plasma cfDNA

An immunological library, plasma technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of not being able to fully and comprehensively display the disease progress of patients, and achieve the removal of randomly introduced errors, The effect of improving accuracy
CN105087789AActive Publication Date: 2015-11-25BEIJING GENEPLUS TECH +2
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
BEIJING GENEPLUS TECH
Publication Date
2015-11-25

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Abstract

The invention provides a method for detecting BCR and TCR immune repertoire in blood plasma cfDNA. The method includes: blood plasma cfDNA extraction, BCR H chain overall length multiple PCR amplification, TCR beta chain CDR3 multiple PCR amplification, PCR amplification after the products of the two previous amplifications are mixed in an equal-volume manner, high-throughput sequencing, and immune repertoire precise information analysis. The primers of the three amplifications are respectively shown in SEQ ID No. 1-28, SEQ ID No. 29-77 and SEQ ID No. 78-79. The invention further provides a kit, which contains the primers, for detecting the BCR and TCR in the blood plasma cfDNA. By the method and the kit which are promising in application prospect in the field of non-invasive detection, the BCR and TCR in the blood plasma cfDNA can be detected at the same time, precise information analysis and precise quantification of immune repertoire subcloning are achieved.
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Description

technical field

[0001] The invention belongs to the technical field of immune repertoire sequencing, in particular to a method for detecting BCR and TCR immune repertoires in plasma cfDNA. Background technique

[0002] Thanks to the rapid development and wide application of next-generation high-throughput sequencing technologies, DNA and RNA sequencing platforms play a prominent role in promoting various aspects of genomics. At the same time, this emerging technical field has also been applied to the sequencing of immune repertoires of T cells and B cell receptors.

[0003] Conventional immune repertoire sequencing of T cells and B cell receptors is mainly based on two major strategies, first: based on cell DNA through TCR / BCR gene rearrangement rules, in TCR (alpha / beta / gamma / delta chain) and BCR The variable region (variable) gene of (heavy / light chain) is set with upstream primers, and the downstream primers are set at the joining region (joining) J gene or constant regi...

Claims

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