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Kit and method for concentrating fetal free DNA, and application of kit

A kit and fetal technology, applied in the field of kits for concentrating fetal free DNA, can solve the problems of false negative test results and low ratio, achieve low gene loss, low cost, and improve diagnostic accuracy

Inactive Publication Date: 2018-12-18
THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem that cffDNA accounts for the low proportion of total free DNA in mother's plasma and easily lead to false negative test results, the present invention provides a kit, method and application for concentrating fetal free DNA, which can detect fetal specific trisomy 21 Chromosome cffDNA as low as 0.1%, showing high sensitivity

Method used

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  • Kit and method for concentrating fetal free DNA, and application of kit
  • Kit and method for concentrating fetal free DNA, and application of kit
  • Kit and method for concentrating fetal free DNA, and application of kit

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Embodiment

[0038] 1.1. Select research objects

[0039] Randomly selected 5 cases of singleton pregnant women with normal pregnancy in May in the Department of Obstetrics and Gynecology of the Second Affiliated Hospital of Soochow University from April 2013 to April 2014. In addition, 3 pregnant women who required induction of labor in 5 months of singleton pregnancy with 21-trisomy syndrome confirmed by karyotype analysis were selected. The pregnant women had no other pregnancy complications and complications. 5ml of peripheral blood was drawn before delivery, and placental tissue was collected during delivery, and DNA extraction was performed immediately.

[0040] 1.2. Method

[0041] 1.2.1. DNA extraction

[0042] Placental tissue was ground into a homogenate, and tissue DNA was extracted according to the instructions (Tiangen DNA Mini Kit, tiangen biotech (Beijing) co., LTD, China), and stored at -80°C. Peripheral blood genomic DNA (Tiangen DNA Mini Kit, Tiangen biotech (Beijing) co., LTD...

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Abstract

The invention relates to a kit and a method for concentrating fetal free DNA, and an application of the kit. DNA extracted from pregnant women peripheral blood or a mixture of placental tissues and the pregnant women peripheral blood is used as a DNA template, MSRE enzyme digestion is carried out, and an amplification primer is adopted to carry out RT-qPCR amplification in order to obtain a PCR product. Compared with kits in the prior art, the kit in the invention has the advantages of detection of the cffDNA of fetal specific trisomy 21st chromosome, being as low as 0.1%, and high sensitivity. The kit is simpler and cheaper than massive parallel sequencing detection of the trisomy 21 syndrome. The kit uses MSRE to process a gene, so the gene loss is less than that of sulfite gene treatment, and the diagnosis accuracy is improved.

Description

Technical field [0001] The invention relates to a kit, method and application for concentrating free fetal DNA. Background technique [0002] Down syndrome is 21-trisomy syndrome, also known as congenital stupidity, usually caused by the appearance of the third chromosome 21. 60% of the children have miscarriages in the early stages of the fetus, and the survivors have obvious mental retardation, special facial features, growth and development disorders and multiple deformities, which bring a heavy economic burden to the family and society. According to the karyotype, children with Down syndrome can be divided into three types: ①Standard type, which accounts for 95% of all cases. The child has 47 somatic chromosomes with an additional 21 chromosome, and the karyotype is 47, XX (or XY), +21. ②Translocation type, accounting for 2.5-5%. The total number of chromosomes in the child is 46, most of which are Robertsonian translocations, with chromosome 14 as the main karyotype 46, X...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12Q1/6883
CPCC12Q1/6806C12Q1/6883C12Q2600/154C12Q2531/113C12Q2521/313
Inventor 张荣肖莉张弘於子扬张书筠李雪徐珍珍
Owner THE SECOND HOSPITAL AFFILIATED TO SUZHOU UNIV
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