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Noninvasive detection method for screening healthily grown blastulas

A detection method, blastocyst technology, applied in the field of epigenetic informatics, can solve problems such as poor accuracy, achieve the effect of improving accuracy and pregnancy success rate

Active Publication Date: 2016-08-17
GUANGZHOU NVWA LIFE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The Gardner morphological blastocyst grading system is a traditional technique that relies on microscopy to select embryos with high morphological grades for transplantation, and the accuracy rate is poor.

Method used

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  • Noninvasive detection method for screening healthily grown blastulas
  • Noninvasive detection method for screening healthily grown blastulas
  • Noninvasive detection method for screening healthily grown blastulas

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Discovery of DNA methylation instability at the genome-wide level in human blastocysts

[0061] The present invention collects blastocysts with the highest grade AA grade and inferior grades CC, BC or CB graded by the Gardner Morphological Blastocyst Grading System. These blastocysts come from the third hospital of Peking University and affiliated to Guangzhou Medical University In the Third Hospital, when a patient is undergoing assisted reproductive surgery, several blastocysts will be cultured in vitro at the same time, and then the one with the best shape will be selected for transplantation. The remaining blastocysts will be donated to the scientific research of this project after signing the informed consent of the patient. Research. For each blastocyst, genome-wide methylation sequencing at single-base resolution was performed. The "one-tube" experimental method of the present invention is as follows:

[0062] 1. Sample preparation

[0063] After the...

Embodiment 2

[0105] Example 2 Incorrect methylation reprogramming affects signaling pathways regulating early embryonic development

[0106] Gene function analysis (gene ontology) was performed on regions with different methylation states in high-grade embryos and low-grade embryos.

[0107] The analysis results found that the differentially methylated regions (DMRs) located in gene promoters are mainly enriched in many pathways related to embryonic development, such as cell cycle, DNA metabolism, DNA modification, chromosome positioning, RNA editing, DNA repair, cell cycle, etc. ( Figure 5 ). These genes whose differentially methylated regions are located inside genes are mainly enriched in the development of specific tissues, such as neural development, neuron differentiation and fate determination, spinal cord structure, mRNA homeostasis, etc. ( Figure 5 ).

[0108] The results also showed that methylation of enhancer regions in the lowest grade CC embryos tended to be erroneously ...

Embodiment 3

[0109] Example 3 The relationship between the methylation level of pre-implantation blastocyst and the birth rate of infants after ART

[0110] In this example, the average methylation level of grade AA blastocysts is 0.3±0.02 as the measurement standard, which is the quantitative standard for non-invasive detection of clinical pre-implantation blastocysts. This standard is based on the morphological detection, and it is a brand-new clinical index for further accurate screening of normally developing blastocysts.

[0111] In order to further verify the accuracy of the index, the present invention has collected 6 blastocysts of B* grade ( figure 2 ), the results showed that the methylation levels of 3 samples in B* grade samples significantly deviated from the standard of 0.3, the average methylation level of AA grade blastocysts. exist figure 1 It shows that 1 of the 5 samples of grade AA deviates from the standard, and 6 of the 7 grade C* embryos tested deviate from the st...

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Abstract

The invention relates to a noninvasive detection method for screening healthily grown blastulas. The method comprises the following steps: the DNA methylation profile of a small amount of trophoblastic cells in the blastulas is detected, the methylation level mean and the methylation pattern of embryos with good morphology in a Gardner morphological blastocyst grading process are adopted as healthily grown blastula screening standards, and embryos well grow and are suitable for being implanted to a maternal when the methylation level of the trophoblastic cells of the blastulas to be detected approaches the methylation level or state of the good blastulas; and a methylation sequencing result is analyzed to judge that whether a chromosome is abnormal or not in order to directly reject chromosome-abnormal embryos. The method makes transplantation of embryos with normal ploidy and excellent apparent genetic maps be clinically realized, greatly improves the accuracy of an assisted reproduction technology, is in favor of reducing the natural abortion rate, improves the pregnancy quality and improves the birth rate of test tube babies.

Description

technical field [0001] The invention relates to the field of epigenetic informatics at the whole genome level, in particular to a non-invasive detection method for screening excellent developing blastocysts. Background technique [0002] In vitro fertilization and embryo transfer (IVF-ET) is commonly known as "test-tube baby". It is a special clinical assisted reproductive technology, which needs to take the eggs and sperm out of the body, let them complete the fertilization process in an artificially controlled environment outside the body, and then transfer the early embryos into the female uterus, where they are bred into child. Babies born using in vitro fertilization are also called test-tube babies. Embryos obtained in vitro should be screened before implantation. Currently, there are two clinical methods: [0003] (1) Gardner morphological blastocyst grading system (Gardner morphological blastocystgrading system). Blastocyst cultivation and transfer are very impor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/154C12Q1/68C12Q1/6881C12Q1/6869C12Q2537/164C12Q2525/191C12Q2525/173C12Q2531/113
Inventor 刘江乔杰李国强于洋范勇李从儒
Owner GUANGZHOU NVWA LIFE TECH CO LTD
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