Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method and kit for detecting existence or proportion of fetal free DNA in pregnant woman sample

A technology for samples and detection probes, applied in recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems of inability to integrate, high sequencing cost, difficult clinical popularization, etc. The effect of high degree of automation and short detection cycle

Pending Publication Date: 2022-06-21
XIAMEN UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the proportion of Y chromosome fragments, the concentration and proportion of cffDNA in male fetuses can be estimated simply and accurately, but it is not applicable to female fetuses; most of the methods for estimating the concentration and proportion of cffDNA on the existing NIPS detection platform are based on the difference in the number and length of cffDNA fragments. The accuracy of the model for fetal FF estimation has yet to be verified by methods such as SNP; while methods such as SNP-based next-generation sequencing can accurately estimate the concentration and ratio of cffDNA, but due to the high cost of sequencing and cannot be integrated into the existing NIPS detection In the process, there is currently no feasibility for large-scale clinical application; the method based on digital PCR can accurately quantify the ratio of cffDNA, but the cost of digital PCR for multiple SNPs and methylation markers at the same time is too high, and it is difficult to popularize in the clinic

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method and kit for detecting existence or proportion of fetal free DNA in pregnant woman sample
  • Method and kit for detecting existence or proportion of fetal free DNA in pregnant woman sample
  • Method and kit for detecting existence or proportion of fetal free DNA in pregnant woman sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0213] Embodiment 1. Selection of candidate SNP sites

[0214] The SNP site of the present application is selected from the single nucleotide polymorphism site library (dbSNP) of the National Center for Biotechnology Information (NCBI) in the United States, and the preferred candidate SNP site meets the following conditions: (1) between different races Fst (population fixation coefficient) 1Mb; (5) In order to avoid linkage between different sites, try to select sites on different chromosomes . The present embodiment selects 23 candidate SNP sites according to the above preferred standards, specifically as shown in Table 1, SNP site information and sequences are queried and downloaded from the dbSNP database of the National Center for Biotechnology Information (NCBI), the alleles The frequency refers to the Asian population frequency from the Thousand Genomes database, and these loci are evenly distributed on each chromosome of the genome.

[0215] Table 1. Candidate SNP s...

Embodiment 2

[0217] Example 2. Detection of Fetal Free DNA Ratio in Noninvasive Prenatal Screening Samples

[0218] In this example, 4 cases of non-invasive prenatal screening samples were collected and detected in the clinic, and the detection ability of the method of the present invention was investigated. The overall experimental process is as follows: figure 2 shown.

[0219] 3.1 Collection and extraction of samples

[0220] Four groups of non-invasive prenatal screening samples were collected, and each group of samples included peripheral blood samples from pregnant women (8-14 weeks of pregnancy) and saliva samples from corresponding parents of the fetus (hereinafter referred to as "mother's sample" and "father's sample" respectively). Saliva samples were collected according to the instructions of the saliva collector (Xiamen Zhishan Biotechnology Co., Ltd., Xiamen), and stored at room temperature. Use EDTA anticoagulant tubes (Zhejiang Gongdong Medical Instrument Co., Ltd., Tai...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present application relates to the field of molecular diagnostics. Specifically, multiple asymmetric PCR amplification and multicolor probe melting curve analysis are utilized to establish a method for detecting SNP loci of a mother-sourced sample and a father-sourced sample. Furthermore, in combination with a digital PCR system, the invention develops a method and a kit for detecting existence or proportion of fetal free DNA in a pregnant woman sample. Compared with the prior art, the method and the kit provided by the invention have the advantages of automatic detection, few manual operation steps, short detection period, accurate detection and high sensitivity.

Description

technical field [0001] This application relates to the field of molecular diagnostics. Specifically, the present application relates to a method for detecting SNP sites in samples derived from the mother and samples derived from the father, and further, relates to a method for detecting the presence or ratio of fetal cell-free DNA in a pregnant woman sample, and a kit . Background technique [0002] As early as 1997, Lo YMD et al. (Lancet.1997; 350:485-487) used PCR technology to detect the specific sequence of the testis-determining gene (SRY) in the plasma and serum of pregnant male fetuses from 12 to 40 weeks of gestation. The presence of fetal free DNA (Cell Free Fetal DNA, cffDNA) was confirmed. The discovery of fetal cell-free DNA has made it possible for non-invasive prenatal diagnosis of monogenic genetic diseases and non-invasive prenatal screening for chromosomal aneuploidy. However, the cell-free DNA obtained in the plasma of pregnant women mainly comes from th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2531/113C12Q2537/143C12Q2527/107C12Q2563/107
Inventor 黄秋英陈昕雯李庆阁
Owner XIAMEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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