Abstract
Fetal nucleated cells circulating in the peripheral blood during pregnancy are potential targets for noninvasive genetic testing. Fluorescence in situ hybridization (FISH) frequently is used to quantify the total number of fetal cells in peripheral blood of pregnant women. We describe an alternative molecular cytogenetic procedure that is the primed in situ labeling (PRINS). This technique consists of annealing oligonucleotides specific to individual chromosome targets and in situ elongation using Taq DNA polymerase to incorporate labeled dUTPs. The sites of the newly synthesized DNA sequences were revealed as fluorescent signals using an immunochemical reaction. The dual-color PRINS was specifically performed for simultaneous detection of two chromosome targets, X and Y. The fluorescent signals corresponding to chromosomes X and Y were displayed as red and green color spots, respectively. The sensitivity and specificity of PRINS are similar to FISH and allow us to efficiently and reliably detect fetal cells in maternal blood. Moreover, dual-color PRINS is faster and more cost-effective than FISH.
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References
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Acknowledgment
The authors are grateful to Dr Kieron Legge for review of the manuscript and to Mr. Marc Bronsard for his technical help. This study was supported by a grant from the Canadian Institute for Health Research and the Canadian Genetic Diseases Network (MRC/NSERC NCE Program) to Régen Drouin. Kada Krabchi was a recipient of a studentship from Valorisation Recherche Québec. Macoura Gadji is a student scholar of the Public Health Minister (MSP) and the National Center of Blood Transfusion (CNTS) of Senegal. Currently, he holds a studentship of the Foundation for Research into Children’s Diseases. Régen Drouin holds the Canada Research Chair in Genetics, Mutagenesis and Cancer.
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Krabchi, K., Gadji, M., Yan, J., Drouin, R. (2006). Dual-Color PRINS for In Situ Detection of Fetal Cells in Maternal Blood. In: Pellestor, F. (eds) PRINS and In Situ PCR Protocols. Methods in Molecular Biology™, vol 334. Humana Press. https://doi.org/10.1385/1-59745-068-5:141
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DOI: https://doi.org/10.1385/1-59745-068-5:141
Publisher Name: Humana Press
Print ISBN: 978-1-58829-549-1
Online ISBN: 978-1-59745-068-3
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