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建立一种实时 PCR 方法用于快速鉴定人类胚胎的性别。

Development of a real-time PCR method for rapid sexing of human preimplantation embryos.

机构信息

CPDP, Paulista Centre of Diagnosis and Research, Ribeirão Preto, Brazil.

出版信息

Reprod Biomed Online. 2010 Jan;20(1):75-82. doi: 10.1016/j.rbmo.2009.10.008. Epub 2009 Oct 30.

Abstract

Genes on the X chromosome are known to be responsible for more than 200 hereditary diseases. After IVF, the simple selection of embryo sex before uterine transfer can prevent the occurrence of affected offspring among couples at risk for these genetic disorders. The aim of this investigation was to develop a rapid method of preimplantation genetic diagnosis (PGD) using real-time polymerase chain reaction (PCR) for the sexing of human embryos, and to compare it to the fluorescence in-situ hybridization technique, considered to be the gold standard. After biopsies were obtained from 40 surplus non-viable embryos for transfer, a total of 98 blastomeres were analysed. It was possible to analyse 24 embryos (60%) by both techniques, generating a total of 70 blastomeres (35 per technique), while 28 blastomeres from 16 embryos (40%) were analysed only by real-time PCR. A rapid and safe method was developed in the present study for the sexual diagnosis of a single human cell (blastomere and buccal cell) using the emerging technology of real-time PCR.

摘要

X 染色体上的基因被认为与 200 多种遗传性疾病有关。在体外受精(IVF)之后,在子宫转移前简单地选择胚胎性别,可以防止有遗传疾病风险的夫妇生育受影响的后代。本研究的目的是开发一种使用实时聚合酶链反应(PCR)对人类胚胎进行性别鉴定的快速方法,即胚胎种植前遗传学诊断(PGD),并将其与荧光原位杂交技术(被认为是金标准)进行比较。在从用于转移的 40 个多余的非存活胚胎中获取活检后,总共分析了 98 个卵裂球。两种技术都可以分析 24 个胚胎(60%),总共生成 70 个卵裂球(每个技术 35 个),而实时 PCR 仅分析了 16 个胚胎中的 28 个卵裂球(40%)。本研究利用实时 PCR 这一新兴技术,开发了一种快速、安全的方法,用于对单个人类细胞(卵裂球和口腔细胞)进行性别诊断。

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