Donor SNP Allele Detection in Transplant Recipient Blood

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Solution Overview

Problem

Current methods for detecting transplant rejection are time-consuming and expensive, and often require DNA from the donor, which may not be available, especially in cases where the transplantation occurred years prior, necessitating a sensitive and rapid technique for early detection.

Innovation Solution

A method involving the monitoring of graft cell-free DNA (cfDNA) by assessing the level of donor-specific single-nucleotide polymorphisms (SNPs) in a recipient's blood sample, without the need for a separate donor sample, using pre-selected SNPs with a minor allele frequency of at least 0.4, and performing quantitative PCR to detect the presence of these SNPs.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If traditional methods are used to detect transplant rejection, then detection accuracy is maintained, but the method becomes extremely time-consuming and expensive

Engineering Contradiction:
Improvedetection accuracyVSAvoidturn-around-time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The invention extracts and utilizes only the necessary genetic information (SNP alleles) from the donor-recipient pair to create a simplified detection method. By focusing on specific SNP markers rather than comprehensive genomic analysis, the method achieves rapid detection without sacrificing accuracy in identifying transplant rejection

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the detection parameters by using SNP allele frequency thresholds (MAF ≥ 0.4) and specific genotyping approaches. This parameter optimization enables the method to achieve both high detection accuracy and rapid turnaround by efficiently identifying donor-specific alleles in recipient samples

Inventive Principle:
Principle #35Parameter changes

2Reliability

If traditional methods are used to detect transplant rejection, then detection reliability is maintained, but the cost becomes prohibitively high

Engineering Contradiction:
Improvedetection reliabilityVSAvoidcost-effectiveness
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The invention employs cost-effective SNP genotyping assays that use inexpensive reagents and can be performed with standard laboratory equipment. The method relies on detecting specific SNP alleles rather than requiring expensive donor DNA samples or complex molecular biology procedures, making the test economically feasible for routine clinical use

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

3Measurement precision

If SNP-based methods are used to differentiate donor and recipient DNA, then detection sensitivity is improved, but the requirement for donor DNA samples increases complexity

Engineering Contradiction:
Improvedetection sensitivityVSAvoidsample requirements
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The invention enables the recipient's own DNA samples to serve dual purposes: first to identify their SNP genotype, and then to detect donor-specific alleles that indicate transplant rejection. This self-referential approach eliminates the need for separate donor DNA samples, reducing sample collection complexity while maintaining high detection sensitivity through SNP allele comparison

Inventive Principle:
Principle #25Self-service

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach allows for early and sensitive detection of transplant rejection, reducing the need for donor DNA and improving turn-around-time and cost-effectiveness, while enabling monitoring of transplant integrity and immunosuppressive therapy adjustments.

Implementation Method 1

performing a quantitative PCR reaction for the SNPs identified in (a) to detect the presence of the alternative allele for one or more of the SNPs

Methodology Applied
Scientific EffectPolymerase Chain Reaction (PCR):

Data Source

PatentEP3004388B2Detection and quantification of donor cell-free DNA in the circulation of organ transplant recipients
Publication Date: 2023.05.31 CHRONIX BIOMEDICAL INC
  • EP3004388B2 patent drawingFigure 1
  • EP3004388B2 patent drawingFigure 2
  • EP3004388B2 patent drawingFigure 3

AI summary

This invention provides methods, compositions, and kits relating to detecting donor cell-free DNA in the circulation of an organ transplant recipient for the early identification of transplant rejection