title: Detection of chromosome aneuploidies in chorionic villus samples by multiplex ligation-dependent probe amplification.
abstract: The objective of this study was to examine the suitability of multiplex ligation-dependent probe amplification (MLPA) in chorionic villus samples as a replacement for traditional karyotyping for the detection of (an)euploidies of chromosomes 21, 18, 13, X, and Y. Chorionic villus samples were diagnosed by traditional karyotyping using short-term cultures (STC) and long-term cultures (LTC), and by MLPA using kit P095. DNA was extracted after digestion of whole villi with proteinase K and/or trypsin and collagenase. Different cell-dissociation procedures were tested to obtain MLPA results representative of the cytotrophoblast layer and the mesenchymal core. Over 95% of the MLPA results were in concordance with the traditional karyotyping of STC and LTC. Traditional karyotyping revealed seven mosaics. After digestion of whole villi with proteinase K, only abnormal cell lines confined to the STC gave rise to abnormal MLPA results. In one sample, the complete discrepancy between STC and LTC was resolved after enzymatic dissociation of cells from the cytotrophoblast layer and the mesenchymal core. MLPA in chorionic villus samples was found to be a reliable test for the detection of (an)euploidies of chromosomes 21, 18, 13, X, and Y. Whole villi digestion with proteinase K resulted in the over-representation of cytotrophoblasts in the DNA pool. To obtain MLPA results representative for STC and LTC, enzymatic dissociation of cells from the cytotrophoblast layer and mesenchymal core is required.
authors.affiliation: Department of Human Genetics, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands. Electronic address: a.kooper@antrg.umcn.nl.
authors.forename: Angelique J A
authors.initials: AJA
authors.lastname: Kooper
authors.affiliation: Department of Human Genetics, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Brigitte H W
authors.initials: BHW
authors.lastname: Faas
authors.affiliation: Departments of Epidemiology, Biostatistics and Health Technology Assessment, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Ton
authors.initials: T
authors.lastname: Feuth
authors.affiliation: Departments of Obstetrics and Gynecology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Johan W T
authors.initials: JWT
authors.lastname: Creemers
authors.affiliation: Department of Obstetrics and Gynecology, Rijnstate Hospital, Arnhem, The Netherlands.
authors.forename: Hans H
authors.initials: HH
authors.lastname: Zondervan
authors.affiliation: Department of Obstetrics and Gynecology, St. Elizabeth's Hospital, Tilburg, The Netherlands.
authors.forename: Peter F
authors.initials: PF
authors.lastname: Boekkooi
authors.affiliation: Department of Obstetrics and Gynecology, Medisch Spectrum Twente, Enschede, The Netherlands.
authors.forename: Rik W P
authors.initials: RWP
authors.lastname: Quartero
authors.affiliation: Department of Obstetrics and Gynecology, Jeroen Bosch Hospital, 's-Hertogenbosch, The Netherlands.
authors.forename: Robbert J P
authors.initials: RJP
authors.lastname: Rijnders
authors.affiliation: Department of Human Genetics, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Ineke
authors.initials: I
authors.lastname: van der Burgt
authors.affiliation: Department of Human Genetics, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Ad Geurts
authors.initials: AG
authors.lastname: van Kessel
authors.affiliation: Department of Human Genetics, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Arie P T
authors.initials: APT
authors.lastname: Smits
mesh_terms: D000782:Aneuploidy; D002824:Chorionic Villi; D002877:Chromosomes, Human; D005260:Female; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D007621:Karyotyping / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D012680:Sensitivity and Specificity