Published online before print December 12, 2008

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Journal of Molecular Diagnostics 2009, Vol. 11, No. 1
Copyright © 2009 American Society for Investigative Pathology & Association for Molecular Pathology
DOI: 10.2353/jmoldx.2009.070140

Detection of Chromosome Aneuploidies in Chorionic Villus Samples by Multiplex Ligation-Dependent Probe Amplification

Angelique J.A. Kooper^*, Brigitte H.W. Faas^*, Ton Feuth, Johan W.T. Creemers, Hans H. Zondervan, Peter F. Boekkooi^¶, Rik W.P. Quartero^||, Robbert J.P. Rijnders^**, Ineke van der Burgt^*, Ad Geurts van Kessel^* and Arie P.T. Smits^*

From the Departments of Human Genetics, * Epidemiology, Biostatistics and Health Technology Assessment, and Obstetrics and Gynecology, Radboud University Nijmegen Medical Center, Nijmegen; the Department of Obstetrics and Gynecology, Rijnstate Hospital, Arnhem; the Department of Obstetrics and Gynecology, ^¶ St. Elizabeth’s Hospital, Tilburg; the Department of Obstetrics and Gynecology, ^|| Medisch Spectrum Twente, Enschede; and the Department of Obstetrics and Gynecology, ** Jeroen Bosch Hospital, ‘s-Hertogenbosch, The Netherlands

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.