Preimplantation genetic diagnosis on embryo by using new single cell nucleic acid amplification technology

Abstract

The invention relates to preimplantation genetic diagnosis on an embryo by using new single cell nucleic acid amplification technology, which mainly utilizes the signal amplification action of the mRNA (messenger Ribose Nucleic Acid) to detect the multiplication or deletion of DNAs (Deoxyribonucleic Acids) of certain chromosome segments. According to the central dogma, the mRNA is transcribed by using the DNA as the template, the abnormity of the number of copies of the DNA template can cause the change of the quantity of the mRNAs; and in the transcription process, the multiplication or deletion of the DNA template can be amplified on the mRNA level, and can be easily detected. The main technical method is as follows: the single cell mRNA of a human embryo is subjected to PCR (Polymerase Chain Reaction) amplification after being subjected to reverse transcription and addition of a common primer; by using the amplification product as the template, quantitative PCR with a 96-pore plate is used for detecting the expression level of 8 genes of a single cell or a small amount of cells; and the detection result can be compared with a normal diploid embryo, so as to distinguish the embryo sex of X chromosome recessive inheritance family history, and the multiplication and deletion of Trisomy 21, Trisomy 18, Trisomy 13 and sex chromosome and carry out genetic diagnosis on some common chromosome anomalies.

Description

Technical field [0001] The new technology can amplify the nucleic acid in the single cell of the human embryo, amplify the nucleic acid signal through PCR amplification, select a specific expression gene, detect the expression of the specific gene of the pre-implantation embryo, and realize the genetic diagnosis of the pre-implantation embryo. Background technique [0002] Preimplantation Genetic Diagnosis (PGD) refers to the preimplantation biopsy and genetic analysis of embryos from patients with genetic risks during in vitro fertilization to select embryos without genetic diseases for implantation into the uterus. cavity, so as to obtain a normal fetal diagnosis method, which can effectively prevent the birth of children with genetic diseases. Preimplantation genetic diagnosis is a new technology developed with the development of human assisted reproductive technology, that is, "test tube baby" technology. It is an extension of prenatal diagnosis and another more promisin...

AI Technical Summary

Problems solved by technology

[0004] The classic method for detecting chromosome doubling or deletion is Fluorescence In Situ Hybridization (FISH), in which DNA probes are labeled with fluorescent dyes of different colors , after hybridization with different chromosomes of blastomere cells fixed on a glass slide, the hybridized part shows fluorescence of different colors under a fluorescent microscope, so as to screen for chromosomal abnormalities, and each probe is relatively expensive

For the FISH method, the probes are monopolized by foreign big medicines, and the probes they use for FISH detection are protected by patents, and there is no independent intellectual property rights in this area in China; each fluorescent dye can only dye one Targets, limited by fluorescent dyes, each blastomere can only use 2 to 5 probes, clinically, three fluorescent dyes are usually used to detect three targets

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