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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0065] Example 1 Detects the design of primers for BCR and TCR in plasma cfDNA
[0066] The immunological polymorphism of BCR is based on the recombination arrangement of V(D)J region fragments. The TCRCDR3 region just covers the TRBV-Junction-TRBD-Junction-TRBJ region, and because of the largest variation, it directly determines the antigen specificity of TCR. Primers were designed for the BCR and TCR gene sequences, aiming to amplify the full-length BCRH chain and the CDR3 region of the TCRBeta chain.
[0067] The primer structure of BCR / TCR-PCR1: from the 5' end to the 3' end, it contains the linker sequence, the specific tag sequence and the upstream and downstream primers for BCR / TCR amplification.
[0068] Design of upstream and downstream primers for BCR amplification: According to the homology analysis of the ORFs of the 7 major families of BCRV genes (FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4), the upstream primers designed with the leader region of the V gene Primers...
Embodiment 2
[0080] Example 2 Establishment of method for detecting BCR and TCR immune repertoire in plasma cfDNA
[0081] 1. Plasma cfDNA extraction:
[0082] 1.1 Plasma Extraction Take 2 tubes (5mL / tube) of peripheral blood from the subject, put them in EDTA anticoagulant tubes, gently turn them upside down (to prevent cell rupture), mix thoroughly 6-8 times, and perform the following within 4-6 hours on the day of blood collection Processing: Centrifuge at 1600g for 10 minutes at 4°C, and divide the supernatant (plasma) into multiple 1.5mL / 2mL centrifuge tubes, and the middle layer of white blood cells cannot be absorbed during the suction process; 16000g at 4°C Centrifuge for 10 minutes to remove residual cells, and transfer the supernatant (plasma) to a new 1.5mL / 2mL centrifuge tube. If the white blood cells cannot be sucked to the bottom of the tube, the required plasma after separation is obtained.
[0083] 1.2 After the plasma sample is processed, the separated plasma and remainin...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com