Journal article
Detection of Increased Amounts of Cell-Free Fetal DNA with Short PCR Amplicons
Clinical chemistry (Baltimore, Md.), v 56(1), pp 136-138
01 Jan 2010
PMID: 19884486
Featured in Collection : UN Sustainable Development Goals @ Drexel
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.
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Details
- Title
- Detection of Increased Amounts of Cell-Free Fetal DNA with Short PCR Amplicons
- Creators
- Aleksandra Sikora - University Hospital of BaselBernhard G. Zimmermann - Fluidigm (United States)Corinne Rusterholz - University Hospital of BaselDaniella Birri - University Hospital of BaselVaraprasad Kolla - University Hospital of BaselOlav Lapaire - University Hospital of BaselIrene Hoesli - University Hospital of BaselVivian Kiefer - University Hospital of BaselLaird Jackson - Drexel UniversitySinuhe Hahn - University Hospital of Basel
- Publication Details
- Clinical chemistry (Baltimore, Md.), v 56(1), pp 136-138
- Publisher
- Amer Assoc Clinical Chemistry
- Number of pages
- 3
- Resource Type
- Journal article
- Language
- English
- Web of Science ID
- WOS:000273466300021
- Scopus ID
- 2-s2.0-73449147027
- Other Identifier
- 991019350683304721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Medical Laboratory Technology