Evaluation of a Novel Assay for Detection of the Fetal Marker RASSF1A: Facilitating Improved Diagnostic Reliability of Noninvasive Prenatal Diagnosis

被引：40

作者：

White, Helen E. ^{[1
,2
]}

Dent, Carolyn L. ^{[1
]}

Hall, Victoria J. ^{[1
]}

Crolla, John A. ^{[3
]}

Chitty, Lyn S. ^{[4
,5
]}

机构：

[1] Salisbury Dist Hosp, Natl Genet Reference Lab Wessex, Salisbury, Wilts, England

[2] Univ Southampton, Fac Med, Southampton SO9 5NH, Hants, England

[3] Salisbury Dist Hosp, Wessex Reg Genet Lab, Salisbury, Wilts, England

[4] UCL, London Inst Child Hlth, Clin & Mol Genet Unit, London, England

[5] Univ Coll London Hosp NHS Fdn Trust, Fetal Med Unit, London, England

来源：

PLOS ONE | 2012年 / 7卷 / 09期

关键词：

MATERNAL PLASMA; DNA; BLOOD; SERUM;

D O I：

10.1371/journal.pone.0045073

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Background: Analysis of cell free fetal (cff) DNA in maternal plasma is used routinely for non invasive prenatal diagnosis (NIPD) of fetal sex determination, fetal rhesus D status and some single gene disorders. True positive results rely on detection of the fetal target being analysed. No amplification of the target may be interpreted either as a true negative result or a false negative result due to the absence or very low levels of cffDNA. The hypermethylated RASSF1A promoter has been reported as a universal fetal marker to confirm the presence of cffDNA. Using methylation-sensitive restriction enzymes hypomethylated maternal sequences are digested leaving hypermethylated fetal sequences detectable. Complete digestion of maternal sequences is required to eliminate false positive results. Methods: cfDNA was extracted from maternal plasma (n = 90) and digested with methylation-sensitive and insensitive restriction enzymes. Analysis of RASSF1A, SRY and DYS14 was performed by real-time PCR. Results: Hypermethylated RASSF1A was amplified for 79 samples (88%) indicating the presence of cffDNA. SRY real time PCR results and fetal sex at delivery were 100% accurate. Eleven samples (12%) had no detectable hypermethylated RASSF1A and 10 of these (91%) had gestational ages less than 7 weeks 2 days. Six of these samples were male at delivery, five had inconclusive results for SRY analysis and one sample had no amplifiable SRY. Conclusion: Use of this assay for the detection of hypermethylated RASSF1A as a universal fetal marker has the potential to improve the diagnostic reliability of NIPD for fetal sex determination and single gene disorders.

引用

页数：5

共 21 条

[1] Free fetal DNA in maternal plasma in anembryonic pregnancies: confirmation that the origin is the trophoblast [J].

Alberry, M. ;

Maddocks, D. ;

Jones, M. ;

Hadi, M. Abdel ;

Abdel-Fattah, S. ;

Avent, N. ;

Soothill, P. W. .

PRENATAL DIAGNOSIS, 2007, 27 (05) :415-418

[2] Hypermethylated RASSF1A in maternal plasma:: A universal fetal DNA marker that improves the reliability of noninvasive prenatal diagnosis [J].

Chan, K. C. Allen ;

Ding, Chunming ;

Gerovassili, Ageliki ;

Yeung, Sze W. ;

Chiu, Rossa W. K. ;

Leung, Tse N. ;

Lau, Tze K. ;

Chim, Stephen S. C. ;

Chung, Grace T. Y. ;

Nicolaides, Kypros H. ;

Lo, Y. M. Dennis .

CLINICAL CHEMISTRY, 2006, 52 (12) :2211-2218

[3] New aids for the non-invasive prenatal diagnosis of achondroplasia: dysmorphic features, charts of fetal size and molecular confirmation using cell-free fetal DNA in maternal plasma [J].

Chitty, L. S. ;

Griffin, D. R. ;

Meaney, C. ;

Barrett, A. ;

Khalil, A. ;

Pajkrt, E. ;

Cole, T. J. .

ULTRASOUND IN OBSTETRICS & GYNECOLOGY, 2011, 37 (03) :283-289

[4] SAFE - The special non-invasive advances in fetal and neonatal evaluation network: aims and achievements [J].

Chitty, Lyn S. ;

van der Schoot, C. Ellen ;

Hahn, Sinuhe ;

Avent, Neil D. .

PRENATAL DIAGNOSIS, 2008, 28 (02) :83-88

[5] Prenatal exclusion of β thalassaemia major by examination of maternal plasma [J].

Chiu, RWK ;

Lau, TK ;

Leung, TN ;

Chow, KCK ;

Chui, DHK ;

Lo, YMD .

LANCET, 2002, 360 (9338) :998-1000

[6] Noninvasive prenatal diagnosis of fetal blood group phenotypes: current practice and future prospects [J].

Daniels, Geoff ;

Finning, Kirstin ;

Martin, Pete ;

Massey, Edwin .

PRENATAL DIAGNOSIS, 2009, 29 (02) :101-107

[7] A non-invasive test for prenatal diagnosis based on fetal DNA present in maternal blood: a preliminary study [J].

Dhallan, Ravinder ;

Guo, Xin ;

Emche, Sarah ;

Damewood, Marian ;

Bayliss, Philip ;

Cronin, Michael ;

Barry, Julie ;

Betz, Jordan ;

Franz, Kara ;

Gold, Katie ;

Vallecillo, Brett ;

Varney, John .

LANCET, 2007, 369 (9560) :474-481

[8] Prenatal treatment of congenital adrenal hyperplasia [J].

Forest, MG ;

Morel, Y ;

David, M .

TRENDS IN ENDOCRINOLOGY AND METABOLISM, 1998, 9 (07) :284-289

[9] Non-invasive prenatal determination of fetal sex: translating research into clinical practice [J].

Hill, M. ;

Finning, K. ;

Martin, P. ;

Hogg, J. ;

Meaney, C. ;

Norbury, G. ;

Daniels, G. ;

Chitty, L. S. .

CLINICAL GENETICS, 2011, 80 (01) :68-75

[10] High-Throughput Droplet Digital PCR System for Absolute Quantitation of DNA Copy Number [J].

Hindson, Benjamin J. ;

Ness, Kevin D. ;

Masquelier, Donald A. ;

Belgrader, Phillip ;

Heredia, Nicholas J. ;

Makarewicz, Anthony J. ;

Bright, Isaac J. ;

Lucero, Michael Y. ;

Hiddessen, Amy L. ;

Legler, Tina C. ;

Kitano, Tyler K. ;

Hodel, Michael R. ;

Petersen, Jonathan F. ;

Wyatt, Paul W. ;

Steenblock, Erin R. ;

Shah, Pallavi H. ;

Bousse, Luc J. ;

Troup, Camille B. ;

Mellen, Jeffrey C. ;

Wittmann, Dean K. ;

Erndt, Nicholas G. ;

Cauley, Thomas H. ;

Koehler, Ryan T. ;

So, Austin P. ;

Dube, Simant ;

Rose, Klint A. ;

Montesclaros, Luz ;

Wang, Shenglong ;

Stumbo, David P. ;

Hodges, Shawn P. ;

Romine, Steven ;

Milanovich, Fred P. ;

White, Helen E. ;

Regan, John F. ;

Karlin-Neumann, George A. ;

Hindson, Christopher M. ;

Saxonov, Serge ;

Colston, Bill W. .

ANALYTICAL CHEMISTRY, 2011, 83 (22) :8604-8610

← 1 2 3 →