Non-invasive Fetal Deafness Gene Detection via Plasma DNA Cyclization
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Solution Overview
Problem
Current methods for detecting fetal deafness pathogenic gene mutations are invasive and cannot be performed non-invasively, making it difficult to accurately diagnose genetic deafness in newborns using venous blood from pregnant women.
Innovation Solution
A method utilizing second-generation high-throughput sequencing technology, specifically the Illumina Miseq sequencer, to detect fetal deafness pathogenic gene mutations by designing primers for GJB2, GJB3, and SLC26A4 genes, connecting plasma DNA with barcode linkers, pre-amplifying, cyclizing, and PCR amplifying the DNA to identify mutations in the fetal DNA present in maternal blood.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If invasive prenatal diagnosis methods are used to detect fetal deafness pathogenic genes, then detection accuracy can be achieved, but the procedure becomes invasive and carries risks to the fetus
Solution Approach 1:
The patent uses maternal plasma as an intermediary medium to access fetal genetic information. Fetal DNA fragments naturally circulate in maternal plasma, allowing indirect detection of fetal deafness pathogenic genes without direct intervention in the fetus. This mediator approach enables non-invasive prenatal diagnosis while maintaining detection accuracy.
Solution Approach 2:
The patent extracts and enriches fetal DNA fragments from maternal plasma through selective amplification using gene-specific primers. By taking out the fetal genetic information from the complex maternal plasma background and concentrating it through PCR amplification, the method achieves accurate detection of pathogenic mutations without invasive procedures.
2Ease of operation
If conventional sequencing methods are used on maternal plasma DNA, then the procedure remains simple, but detection sensitivity is insufficient due to low fetal DNA concentration
Solution Approach 1:
The patent performs preliminary enrichment of fetal DNA fragments before sequencing by using gene-specific primers to selectively amplify target regions in maternal plasma DNA. This preliminary action concentrates the low-abundance fetal genetic material, making it detectable by conventional sequencing methods while maintaining procedural simplicity.
Solution Approach 2:
The patent employs a universal amplification strategy using gene-specific primers that can detect multiple deafness-related genes (GJB2, GJB3, SLC26A4) simultaneously from maternal plasma. This multi-functional approach enables detection of various pathogenic mutations across different genes using a single standardized protocol, maintaining simplicity while improving sensitivity.
Data Source
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AI summary
The present invention is directed to a method, kit and primers for detecting fetal deafness pathogenic gene mutations. The method of the invention comprises: (a) designing primers according to the pre-determined mutation loci of deafness pathogenic genes; (b) extracting plasma DNAs in a pregnant woman; (c) connecting the extracted plasma DNAs with pre-amplification linkers to obtain connected products; (d) PCR pre-amplifying the connected product to obtain pre-amplified products; (e) cyclizing the pre-amplified products to obtain cyclised DNAs; (f) PCR amplifying the cyclised DNAs using the designed primers to obtain amplified products; and (g) high throughput sequencing the amplified products and analyzing the mutations of the fetal deafness pathogenic genes. The invention can effectively determine whether the pre-determined loci on deafness pathogenic genes have been mutated as well as the mutation type.