Selection of embryo of test tube baby through sequencing by single cell genome of polar body or embryo

A genome and single-cell technology, applied in the direction of microbial measurement/testing, biochemical equipment and methods, etc., can solve problems such as inability to diagnose

Inactive Publication Date: 2014-08-27
HARVARD UNIV +2
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Problems solved by technology

Array CGH can only diagnose changes in the number of chromosomes and large fragments of chromosome structure (unbalanced translocation, duplication, deletion), and SNP array can perform all chromosomal abnormalities that CGH can diagnose. The resolution is high

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  • Selection of embryo of test tube baby through sequencing by single cell genome of polar body or embryo
  • Selection of embryo of test tube baby through sequencing by single cell genome of polar body or embryo
  • Selection of embryo of test tube baby through sequencing by single cell genome of polar body or embryo

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Embodiment

[0121] The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent manufacturers. Quantitative experiments in the following examples were all set up to repeat the experiments three times, and the results were averaged.

[0122] Part 1: A method for genome sequencing using the first polar body and the second polar body and the female pronucleus of the embryo, including the following steps:

[0123] 1. The process of obtaining the first polar body, the second polar body and the female pronucleus of the oocyte:

[0124] At 1.0: microinjection of single sperm (ICSI) into eggs at stage MII, put the eggs into (G-MOPS) operating fluid, transfer to the platform of the micromanipulator, and p...

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Abstract

Disclosed is a method for using a first polar body and a second polar body as well as a single-cell embryo to carry out whole-genome non-exponential amplification and high-throughput genome sequencing, so as to perform preimplantation genetic diagnosis for genetic disease and testing for pathogenic genes causing repeated miscarriages. The method of the present invention comprises the following steps: (1) obtaining oocytes and embryos, and carrying out separation and genome amplification of first and second polar bodies and single-cell embryos; (2) establishing a genome sequencing library and sequencing, and carrying out bioinformatic analysis of the genome to obtain a gene spectrum and information concerning the number of copies of chromosomes and fragments thereof; (3) determining the chromosome ploidy of the polar bodies and embryos as well as information concerning defects, replication and point mutation in the chromosome fragments; (4) selecting normal or suitable embryos for implantation.

Description

technical field [0001] The invention relates to a method for selecting embryos for test-tube babies by utilizing the first polar body and the second polar body or genome amplification and high-throughput sequencing of embryonic single cells. Background technique [0002] With the rapid development of reproductive medicine and test-tube baby technology, in vitro fertilization-embryo transfer (IVF-ET) and its related derivative technologies have become the most important means of infertility treatment. It is estimated that there are currently about 60-80 million infertile couples in the world, and about 12-15 million infertile couples in China. According to data in 2010, more than 4 million test-tube babies have been born in the world. According to reports, 146,200 cycles of ART were administered in the United States in 2009; in China, about 300,000 couples chose assisted reproductive technology for infertility treatment in 2012 alone. Generally speaking, the current IVF-ET d...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/68C12Q1/6858C12Q1/6806C12Q1/6809C12Q1/6869
Inventor 谢晓亮乔杰汤富酬闫丽盈樊伟侯宇
Owner HARVARD UNIV
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