Expert Review 01
Introduction to QF-PCR by Dr. Kathy Mann
This paper provides straightforward advice for laboratory professionals on how to operate a high-quality QF-PCR service — from basic set up to best practice guidelines.
Authored by Dr. Kathy Mann, a pioneer in prenatal QF-PCR analysis, it includes useful sections on: assay design, sample processing, limitations and much more.
What you'll learn
- Principle of QF-PCR and its advantages
- Why the availability of commercial QF-PCR assays has proven to be a welcome development
- Best practice guidelines for QF-PCR
- Factors that can lead to incidences of samples where no QF-PCR result is possible
- What the future holds for prenatal screening
QF-PCR analysis exploits the variability of microsatellite sequences (STRs), that vary in length at any given locus to distinguish between chromosome homologues. The detection and quantitation of STR alleles enables relative chromosome quantitation (see Figure 1).
One of the advantages of QF-PCR over other approaches for rapid aneuploidy detection is its ability to identify features associated with prenatal samples such as mosaicism and maternal cell contamination. Analysis of the sample genotype is able to identify if more than one cell line is present which may indicate sample mosaicism or maternal cell contamination or could represent twin, chimeric or external sources of DNA; mosaicism and maternal cell contamination give characteristic allele patterns and can usually be confidently categorised (see Expert Review 02: Detecting mosaicism with QF-PCR and Expert Review 03: QF-PCR and maternal cell contamination). The genotype can also be used for sample identification by comparison with a second aliquot of the original sample or a maternal or paternal genotype.