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Detection of increased amounts of cell-free fetal DNA with short PCR amplicons

Sikora, Aleksandra and Zimmermann, Bernhard G. and Rusterholz, Corinne and Birri, Daniella and Kolla, Varaprasad and Lapaire, Olav and Hoesli, Irene and Kiefer, Vivian and Jackson, Laird and Hahn, Sinuhe. (2010) Detection of increased amounts of cell-free fetal DNA with short PCR amplicons. Clinical chemistry, Vol. 56, no. 1. pp. 136-138.

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Official URL: http://edoc.unibas.ch/dok/A6006196

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Abstract

AIM: A digital PCR approach has recently been suggested to detect greater amounts of cell-free fetal DNA in maternal plasma than conventional real-time quantitative PCR (qPCR). Because the digital qPCR approach uses shorter PCR amplicons than the real-time qPCR assay, we investigated whether a real-time qPCR assay appropriately modified for such short amplicons would improve the detection of cell-free fetal DNA. METHOD: We developed a novel universal-template (UT) real-time qPCR assay that was specific for the DYS14 sequence on Y chromosome and had a short amplicon size of 50 bp. We examined this "short" assay with 50 maternal plasma samples and compared the results with those for a conventional real-time qPCR assay of the same locus but with a longer amplicon (84 bp). RESULTS: Qualitatively, both assays detected male cell-free fetal DNA with the same specificity and detection capability. Quantitatively, however, the new UT real-time qPCR assay for shorter amplicons detected, on average, almost 1.6-fold more cell-free fetal DNA than the conventional real-time qPCR assay with longer amplicons. CONCLUSIONS: The use of short PCR amplicons improves the detection of cell-free fetal DNA. This feature may prove useful in attempts to detect cell-free fetal DNA under conditions in which the amount of template is low, such as in samples obtained early in pregnancy.
Faculties and Departments:03 Faculty of Medicine > Bereich Spezialfächer (Klinik) > Gynäkologie und Geburtshilfe > Gynäkologie (Heinzelmann)
03 Faculty of Medicine > Departement Klinische Forschung > Bereich Spezialfächer (Klinik) > Gynäkologie und Geburtshilfe > Gynäkologie (Heinzelmann)
03 Faculty of Medicine > Bereich Spezialfächer (Klinik) > Gynäkologie und Geburtshilfe > Geburtshilfe (Hösli-Krais)
03 Faculty of Medicine > Departement Klinische Forschung > Bereich Spezialfächer (Klinik) > Gynäkologie und Geburtshilfe > Geburtshilfe (Hösli-Krais)
UniBasel Contributors:Hösli-Krais, Irene M. and Lapaire-Mayer, Olav Carl
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:American Association for Clinical Chemistry
ISSN:0009-9147
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:26 Apr 2013 07:02
Deposited On:26 Apr 2013 06:57

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