Facilitating the Fragile X post- and prenatal genetic diagnostic testing workflow through the Abbott FraXa TP-PCR and sizing PCR.

Onderzoeksoutput: Meeting abstract (Book)

Samenvatting

Molecular testing of FMR1 (CGG)n expanded repeats remains hard to tackle given the limitations of sizing PCR in detecting large and uninformative alleles, resulting in a significant number of samples that require confirmation through laborious southern blot analysis. Abbott recently developed a TP-PCR in combination with a sizing PCR to facilitate the Fragile X diagnostic testing workflow.
This study aimed at comparing Abbott FraXa sizing PCR and TP-PCR versus in-house sizing PCR and southern blot in order to evaluate the sizing accuracy and detection sensitivity of normal, intermediate, premutation and mutation alleles. Over 100 samples of different sources (approximately 50% whole blood, 40% chorion villi and 10% amniocytes) and a panel of artificially mimicked mosaic samples were evaluated.
Signal intensity and sizing accuracy met expectations in the normal to small premutation range with a deviation of ± 1 repeat regardless the sample source or DNA extraction method used. The sizing 'long run' greatly improved the detection capacity and sizing accuracy of longer range (large premutation to full mutation) fragments, although inspection of raw data is recommended. TP-PCR allowed discrimination between normal/intermediate and premutation/mutation alleles. However, premutation and mutation TP-PCR signal patterns were very similar, therefore requiring the Abbott sizing PCR to distinguish them.
Mosaic detection in sizing- and TP-PCR ranged from 20% to 10% depending on the repeat range mixture used.
Our results corroborate the suggested Abbott workflow of using FraXa TP-PCR as a first line screening platform in combination with the sizing PCR as second line test.
Originele taal-2English
TitelAbstract book of the 12th Annual Meeting of the Belgian Society of Human Genetics
Pagina's92-92
Aantal pagina's1
StatusPublished - 2 mrt 2012

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