微流控数字PCR技术可实现胎儿非整倍体的快速产前诊断。
Microfluidic digital PCR enables rapid prenatal diagnosis of fetal aneuploidy.
作者信息
Fan H Christina, Blumenfeld Yair J, El-Sayed Yasser Y, Chueh Jane, Quake Stephen R
机构信息
Department of Bioengineering, Stanford University and Howard Hughes Medical Institute, Stanford, CA, USA.
出版信息
Am J Obstet Gynecol. 2009 May;200(5):543.e1-7. doi: 10.1016/j.ajog.2009.03.002.
Abstract
OBJECTIVE
The purpose of this study was to demonstrate that digital polymerase chain reaction (PCR) enables rapid, allele independent molecular detection of fetal aneuploidy.
STUDY DESIGN
Twenty-four amniocentesis and 16 chorionic villus samples were used for microfluidic digital PCR analysis. Three thousand and sixty PCR reactions were performed for each of the target chromosomes (X, Y, 13, 18, and 21), and the number of single molecule amplifications was compared to a reference. The difference between target and reference chromosome counts was used to determine the ploidy of each of the target chromosomes.
RESULTS
Digital PCR accurately identified all cases of fetal trisomy (3 cases of trisomy 21, 3 cases of trisomy 18, and 2 cases of triosmy 13) in the 40 specimens analyzed. The remaining specimens were determined to have normal ploidy for the chromosomes tested.
CONCLUSION
Microfluidic digital PCR allows detection of fetal chromosomal aneuploidy utilizing uncultured amniocytes and chorionic villus tissue in less than 6 hours.
摘要
目的
本研究的目的是证明数字聚合酶链反应（PCR）能够实现对胎儿非整倍体的快速、不依赖等位基因的分子检测。
研究设计
24份羊水样本和16份绒毛膜绒毛样本用于微流控数字PCR分析。对每个目标染色体（X、Y、13、18和21）进行3060次PCR反应，并将单分子扩增的数量与参考值进行比较。目标染色体计数与参考染色体计数之间的差异用于确定每个目标染色体的倍性。
结果
在分析的40个样本中，数字PCR准确识别出所有胎儿三体病例（3例21三体、3例18三体和2例13三体）。其余样本被确定在所检测的染色体上具有正常倍性。
结论
微流控数字PCR能够利用未培养的羊水细胞和绒毛膜绒毛组织在不到6小时内检测胎儿染色体非整倍体。
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本文引用的文献
1
Rapid prenatal diagnosis using uncultured amniocytes and oligonucleotide array CGH.使用未培养的羊水细胞和寡核苷酸阵列比较基因组杂交技术进行快速产前诊断。
Prenat Diagn. 2008 Oct;28(10):943-9. doi: 10.1002/pd.2087.
2
Studying copy number variations using a nanofluidic platform.使用纳米流体平台研究拷贝数变异。
Nucleic Acids Res. 2008 Oct;36(18):e116. doi: 10.1093/nar/gkn518. Epub 2008 Aug 18.
3
Multiplex ligation-dependent probe amplification versus karyotyping in prenatal diagnosis: the M.A.K.E. study.多重连接依赖探针扩增技术与核型分析在产前诊断中的比较：M.A.K.E. 研究
BMC Pregnancy Childbirth. 2008 May 20;8:18. doi: 10.1186/1471-2393-8-18.
4
ACOG Practice Bulletin No. 88, December 2007. Invasive prenatal testing for aneuploidy.美国妇产科医师学会实践公告第88号，2007年12月。非整倍体的侵入性产前检测。
Obstet Gynecol. 2007 Dec;110(6):1459-67. doi: 10.1097/01.AOG.0000291570.63450.44.
5
Detection of aneuploidy with digital polymerase chain reaction.利用数字聚合酶链反应检测非整倍体
Anal Chem. 2007 Oct 1;79(19):7576-9. doi: 10.1021/ac0709394. Epub 2007 Aug 24.
6
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Proc Natl Acad Sci U S A. 2007 Aug 7;104(32):13116-21. doi: 10.1073/pnas.0705765104. Epub 2007 Jul 30.
7
Array-CGH analysis of cell-free fetal DNA in 10 mL of amniotic fluid supernatant.对10毫升羊水上清液中游离胎儿DNA进行阵列比较基因组杂交分析。
Prenat Diagn. 2007 Jul;27(7):616-21. doi: 10.1002/pd.1752.
8
Aneuploidy in the miscarriages of infertile women and the potential benefit of preimplanation genetic diagnosis.不孕女性流产中的非整倍体及植入前基因诊断的潜在益处。
Fertil Steril. 2008 Feb;89(2):353-7. doi: 10.1016/j.fertnstert.2007.02.040. Epub 2007 May 16.
9
Molecular cytogenetic and rapid aneuploidy detection methods in prenatal diagnosis.产前诊断中的分子细胞遗传学和快速非整倍体检测方法。
Am J Med Genet C Semin Med Genet. 2007 Feb 15;145C(1):87-98. doi: 10.1002/ajmg.c.30114.
10
Microfluidic digital PCR enables multigene analysis of individual environmental bacteria.微流控数字PCR可对单个环境细菌进行多基因分析。
Science. 2006 Dec 1;314(5804):1464-7. doi: 10.1126/science.1131370.
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