Almasi Mohammad Amin, Almasi Galavizh
- Young Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran.
J Reprod Infertil. 2017 Jan-Mar;18(1):197-204.
In human, SRY (sex-determining region of the Y chromosome) is the major gene for the testis-determining factor which is found in normal XY males and in the rare XX males, and it is absent in normal XX females and many XY females. There are several methods which can indicate a male genotype by the presence of the amplified product of SRY gene. The aim of this study was to identify the SRY gene for embryo sex determination in human during pregnancy using loop mediated isothermal amplification (LAMP) method.
A total of 15 blood samples from pregnant women at eight weeks of pregnancy were collected, and Plasma DNA was extracted. LAMP assay was performed using DNA obtained for detection of SRY gene. Furthermore, colorimetric LAMP assay for rapid and easy detection of SRY gene was developed.
LAMP results revealed that the positive reaction was highly specific only with samples containing XY chromosomes, while no amplification was found in samples containing XX chromosomes. A total of 15 blood samples from pregnant women were seven male embryos (46.6%) and eight female embryos (53.4%). All used visual components in the colorimetric assay could successfully make a clear distinction between positive and negative ones.
The LAMP assay developed in this study is a valuable tool capable of monitoring the purity and detection of SRY gene for sex determination.
在人类中，SRY（Y染色体性别决定区）是睾丸决定因子的主要基因，存在于正常XY男性以及罕见的XX男性中，而在正常XX女性和许多XY女性中不存在。有几种方法可通过SRY基因扩增产物的存在来指示男性基因型。本研究的目的是使用环介导等温扩增（LAMP）方法鉴定人类孕期胚胎性别鉴定中的SRY基因。
收集了15例妊娠8周孕妇的血样，并提取了血浆DNA。使用获得的DNA进行LAMP分析以检测SRY基因。此外，还开发了用于快速简便检测SRY基因的比色LAMP分析。
LAMP结果显示，阳性反应仅对含XY染色体的样本具有高度特异性，而含XX染色体的样本未发现扩增。15例孕妇血样中，有7例为男性胚胎（46.6%），8例为女性胚胎（53.4%）。比色分析中使用的所有视觉成分均可成功区分阳性和阴性。
本研究开发的LAMP分析是一种有价值的工具，能够监测用于性别鉴定的SRY基因的纯度和检测。
