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Kit, device and method for increasing fetal free dna concentration in maternal peripheral blood

A peripheral blood, high-concentration technology, used in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problems of missed detection of positive samples, false negatives, etc. Concentration, cost reduction effect

Active Publication Date: 2018-09-07
CAPITALBIO GENOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For NIPT, a low concentration of fetal cell-free DNA increases the risk of missing a positive sample
The detection limit of NIPT is 3% to 4%. However, due to the influence of individual differences in pregnant women, sample quality, and experimental errors, in the prior art, when the concentration of fetal free DNA is lower than 5%, the NIPT test results may be false negative.

Method used

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  • Kit, device and method for increasing fetal free dna concentration in maternal peripheral blood
  • Kit, device and method for increasing fetal free dna concentration in maternal peripheral blood
  • Kit, device and method for increasing fetal free dna concentration in maternal peripheral blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1: A kind of kit that improves fetal cell-free DNA concentration in maternal peripheral blood

[0066] The kit consists of free DNA extraction reagents, library building reagents and free DNA enrichment reagents, among which the free DNA enrichment reagents can enrich free DNA fragments at any point or any segment of free DNA with a fragment length not exceeding 160bp .

[0067] Specifically, the free DNA extraction reagent is a magnetic bead extraction reagent, including proteinase K, lysis buffer, cleaning solution 1, cleaning solution 2, and eluent.

[0068] Library building reagents are library building reagents for fetal chromosomal aneuploidy, including: end repair buffer, end repair enzyme, DNA ligase, DNA ligase, PCR amplification reagent, PCR primers, TE solution, P1 adapter, label Sequences 1-32; Ion Plus Fragment Library Ki (Life Technology) or New England Biolabs, and related library construction reagents from Enzymatics can also be used. In thi...

Embodiment 2

[0117] Embodiment 2: A kind of test kit that improves fetal cell-free DNA concentration in maternal peripheral blood

[0118] The composition of the kit is the same as in Example 1.

[0119] Samples and their processing: The samples come from 10ml maternal venous blood samples of Beijing Boao Medical Laboratory, numbered "RD13120110", "RD13120111", "RD13120112" and "RD13120113". These samples were all collected from the peripheral blood of pregnant women who were pregnant with male fetuses. Among them, RD13120112 was a T21-positive sample, and the other three samples were confirmed as normal fetuses by clinical follow-up. Blood samples were centrifuged in two steps to obtain blood cell-depleted plasma, which could be used for subsequent experiments or frozen at -80°C for future use.

[0120]Extract plasma free DNA: select the serum / plasma free DNA magnetic bead extraction kit (product number S10020) produced by Boao Muhua Company for DNA extraction, extract three copies of DN...

Embodiment 3

[0147] Embodiment 3: A kind of device that improves fetal cell-free DNA concentration in maternal peripheral blood

[0148] Obtain short fragments of free DNA <160bp in a certain section or a certain point in maternal peripheral blood by physical methods, and the devices for separating short fragments of free DNA by physical methods are gel electrophoresis, prefabricated gel electrophoresis, etc., including electrophoresis tanks, gels, DNAladder, electrophoresis platform and operating system.

[0149] The device includes a peripheral blood free DNA separation device, a free DNA library construction device, a library purification device and a free DNA enrichment device for enriching free DNA fragments at any point or segment of free DNA whose fragment length does not exceed 160 bp. in:

[0150] The peripheral blood cell-free DNA separation device is a magnetic bead extraction kit, including: proteinase K, lysis buffer, cleaning solution 1, cleaning solution 2, and eluent;

[...

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Abstract

The invention discloses a kit, device and method for improving a concentration of fetal free DNA in maternal peripheral blood. A lot of studies prove that through sequencing and data analysis of enriched free DNA at a point or in any region less than 160 bp in maternal peripheral blood, free DNA in maternal peripheral blood can be unexpectedly enriched and a concentration (%) of fetal free DNA isimproved by 1.5-4 times. The average concentration of the fetal free DNA in maternal peripheral blood is 13%. The device and method can improve the average concentration of the fetal free DNA to 20-40% and have remarkable effects. Through effectively increasing the fetal free DNA concentration, the data processing capacity of high-energy sequencing is greatly reduced and a high-energy sequencing cost is further reduced.

Description

technical field [0001] The invention relates to a pretreatment kit, device and method containing free DNA samples, in particular to a kit, device and method for increasing the concentration of fetal free DNA in maternal peripheral blood. Background technique [0002] The incidence of birth defects caused by fetal chromosomal abnormalities (including chromosomal aneuploidy and microdeletion and microduplication abnormalities) is about 1%. Chromosomal abnormalities mainly refer to abnormalities in chromosome number and structure, including trisomy 21 (T21), 18 - Trisomy (T18), Trisomy 13 (T13), XO syndrome, XXX syndrome, XYY syndrome and XXY syndrome and various microdeletion and microduplication syndromes. Most of the children are mentally handicapped, developmental delay, multiple deformities, or fertility disorders in adulthood. [0003] Single-gene genetic disease refers to a genetic disease controlled by a pair of alleles. According to the statistics of the online Mendel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6806C12M1/00
Inventor 糜庆丰陈样宜黄铨飞彭春方罗东红饶兴蔷
Owner CAPITALBIO GENOMICS
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