Multiplex NGS Genotyping for 17 RBC Blood Group Systems
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Solution Overview
Problem
Current blood transfusion practices face challenges in accurately matching unconventional blood group systems, leading to adverse reactions and the scarcity of rare blood groups due to limitations in existing detection technologies, particularly serological tests and low-throughput genetic methods like PCR-SSP and Sanger sequencing, which are costly and inefficient for screening rare blood groups.
Innovation Solution
A method and kit utilizing next-generation sequencing (NGS) with a multiplex PCR-based primer set for simultaneously detecting 17 RBC blood group systems, including primers with specific nucleotide sequences, to enhance accuracy and efficiency in genotyping, suitable for Illumina sequencing platforms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If serological test is used for blood group identification, then operation is simple and rapid, but detection of unconventional blood group systems is limited due to unavailability of antibody reagents
Solution Approach 1:
The patent replaces the serological test method (which relies on physical/chemical reactions between antibodies and antigens) with a genetic detection method based on PCR and sequencing. This substitution allows detection of blood group systems regardless of antibody reagent availability, as it directly analyzes the genetic determinants of blood group antigens.
Solution Approach 2:
The patent develops a universal genetic detection platform that can identify multiple blood group systems (ABO, Rh, Kidd, Duffy, MNS, and others) through a single methodology. The primer set and sequencing approach provide multi-functional capability to detect both conventional and unconventional blood group systems that serological tests cannot cover.
2Measurement precision
If PCR-SSP or Sanger sequencing is used for blood group genotyping, then genetic detection capability is achieved, but detection efficiency and cost-effectiveness are insufficient for large-scale screening
Solution Approach 1:
The patent combines multiple low-throughput methods (PCR amplification with specific primers and Sanger sequencing) into a unified high-throughput NGS platform. By merging the specificity of PCR-based approaches with the parallel processing capability of next-generation sequencing, the system achieves both accurate genotyping and high detection throughput suitable for large-scale screening.
Solution Approach 2:
The patent changes the key parameter of detection throughput by transitioning from traditional sequencing methods to next-generation sequencing technology. This parameter change enables simultaneous analysis of multiple samples and multiple blood group systems, dramatically increasing productivity while maintaining genotyping accuracy.
3Reliability
If conventional blood group testing is performed, then routine transfusion safety is ensured, but rare blood groups cannot be identified leading to supply shortages
Solution Approach 1:
The patent performs preliminary genetic identification of rare blood groups in the donor population before clinical need arises. By proactively screening donors using the high-throughput NGS method, the system identifies and can prioritize rare blood group donors, preventing supply shortages when clinically needed without compromising routine transfusion safety.
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
The NGS-based method improves detection efficiency and accuracy for RBC blood group genotyping, allowing for high-throughput screening of rare blood groups, reducing costs, and providing comprehensive and accurate detection results, including hereditary mutations and unknown mutation sites.
Implementation Method 1
a first amplification is performed separately in three capture and amplification systems to obtain three first amplification products, wherein the three capture and amplification systems are respectively constructed by using a nucleic acid extracted from a sample as a template and a first primer set, a second primer set, and a third primer set
Implementation Method 2
based on next-generation sequencing (NGS)... High-throughput gene sequencing is conducted to simultaneously detect 17 RBC blood group system-related genes
Data Source
AI summary
A method and a kit for genotyping of multi-system red blood cell (RBC) blood group based on next-generation sequencing (NGS) are provided. Multiplex PCR capture primer sets and amplification reaction conditions thereof can be designed using Illumina as a platform to simultaneously detect full-length coding regions and spliced flanking regions of 25 blood group genes in 17 RBC blood group systems among three reactions, thereby enabling rapid genotyping of the 17 RBC blood group systems and multiple key rare blood groups therein. The method and the kit are based on high-throughput sequencing and have a high detection efficiency and a low average cost; the method and the kit also break through a limitation of existing blood group gene detection technology that cannot distinguish haplotypes; moreover, variations including known mutation sites and unknown mutation sites can be detected to obtain comprehensive and accurate detection results.