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MeDIP real‐time qPCR of maternal peripheral blood reliably identifies trisomy 21
Author(s) -
Tsaliki Evdokia,
Papageorgiou Elisavet A.,
Spyrou Christiana,
Koumbaris George,
Kypri Elena,
Kyriakou Skevi,
Sotiriou Chrysovalanto,
Touvana Evi,
Keravnou Anna,
Karagrigoriou Alex,
Lamnissou Klea,
Velissariou Voula,
Patsalis Philippos C.
Publication year - 2012
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/pd.3947
Subject(s) - methylated dna immunoprecipitation , differentially methylated regions , trisomy , cell free fetal dna , peripheral blood , real time polymerase chain reaction , computational biology , computer science , biology , prenatal diagnosis , medicine , dna methylation , fetus , pregnancy , genetics , gene , gene expression
ABSTRACT Objective To reevaluate the efficiency of the 12 differentially methylated regions (DMRs) used in the methylated DNA immunoprecipitation (MeDIP) real‐time quantitative polymerase chain reaction (real‐time qPCR) based approach, develop an improved version of the diagnostic formula and perform a larger validation study. Methods Twelve selected DMRs were checked for copy number variants in the Database of Genomic Variants. The DMRs located within copy number variants were excluded from the analysis. One hundred and seventy‐five maternal peripheral blood samples were used to reconstruct and evaluate the new diagnostic formula and for a larger‐scale blinded validation study using MeDIP real‐time qPCR. Results Seven DMRs entered the final model of the prediction equation and a larger blinded validation study demonstrated 100% sensitivity and 99.2% specificity. No significant evidence for association was observed between cell free fetal DNA concentration and D value. Conclusion The MeDIP real‐time qPCR method for noninvasive prenatal diagnosis of trisomy 21 was confirmed and revalidated in 175 samples with satisfactory results demonstrating that it is accurate and reproducible. We are currently working towards simplification of the method to make it more robust and therefore easily, accurately, and rapidly reproduced and adopted by other laboratories. Nevertheless, larger scale validation studies are necessary before the MeDIP real‐time qPCR‐based method could be applied in clinical practice. © 2012 John Wiley & Sons, Ltd.