甲基化荧光PCR定量检测母体循环中不同妊娠状态下胎儿maspin基因  

The value of the detection of fetal U-Maspin gene in maternal plasma at different pregnant states by real-time fluorescence quantitative PCR

作　　者：汤冬玲[1] 魏新素[1] 

机构地区：[1]武汉大学人民医院检验科,武汉430060

出　　处：《中国优生与遗传杂志》2013年第10期143-144,91,共3页Chinese Journal of Birth Health & Heredity

基　　金：国家自然基金(81102141);湖北省自然科学基金(2009CDB194);湖北省卫生厅青年人才基金(QJX2010-15)

摘　　要：目的依据孕妇血浆中存在游离胎儿DNA的理论,从孕妇外周血浆中分离出胎儿DNA并加以鉴定,预防X连锁遗传病患儿的出生。方法从孕早期、中期及晚期共80名孕妇外周血浆中分离胎儿DNA,采用实时荧光定量PCR(real-time fluorescence quantitative polymerase chain-reaction,FQ-PCR)方法对不同妊娠状态下孕妇外周血中游离的胎儿DNA(fDNA)进行定量分析,初步探讨游离fDNA在不同妊娠状态下的浓度变化,为尽早应用于临床提供科学依据。结果早孕组28例血浆标本中有25例检测到U-MASPIN基因,其平均浓度为57.82拷贝数/ml;中孕组20例血浆标本中均检测到U-maspin基因,其平均浓度为153.08拷贝数/ml;晚孕组32例血浆标本中均检测到U-maspin基因,其平均浓度为219.34拷贝数/ml,组间两两比较,差异有极显著意义(P<0.01)。结论用实时荧光定量PCR的方法最早在孕72天的孕妇外周血浆中就可以检测到胎儿U-maspin基因,随孕周的增加,母血中胎儿DNA的量也在逐渐增加。实时荧光定量PCR技术在进行无创伤性产前性别诊断中有重要的价值。Objective： To investigate the feasibility of quantitative detection of expression of maspin gene methylated （ M - maspin ） and unmethylated （ U - maspin ） in maternal plasma with methylation - specific PCR. Methods ： Fetal DNA in maternal plasma was isolated from 28 cases bearing fetuses in the first trimester, 20 in the mid - trimester of pregnancy and 32 in the late - trimester of pregnancy, respectively. Real -time Fluorescence Quantitative Polymerase Chain -reaction （FQ -PCR） was used to determine the expressions of M - maspin and U - maspin gene. Free DNA of plasma samples was extracted, then a standard curve using serial dilutions of the standard preparation was employed. Results ： It shows The expression of U - maspin gene in maternal plasma of three pregnant women at the first trimester of pregnancy was not detected. The mean concentrations of U - maspin gene were 57.82 copies per milliliter in the first trimester, 153.08 copies per milliliter in the second trimester, and 219.34 copies per milliliter in the third trimester. Conclusion ： The results show that U - maspin gene can be found as early as 72 days of gestation in maternal plasma by the use of FQ - PCR. U - maspin gene may be considered as an epigenetic marker to detect the fetal DNA in maternal plasma. It is easy and precise to detect the microamount of U - maspin gene in maternal plasma by methylight technology, has clear potential for clinical application in non- invasive prenatal diagnosis.

关 键 词：实时荧光定量聚合酶链反应 母体血浆 U—maspin 

分 类 号：R714.5[医药卫生—妇产科学]