Implementing Massive Parallel Pyrosequencing in Molecular Diagnostics
Author Information
Author(s): Kim De Leeneer, Joachim De Schrijver, Lieven Clement, Machteld Baetens, Steve Lefever, Sarah De Keulenaer, Wim Van Criekinge, Dieter Deforce, Filip Van Nieuwerburgh, Sofie Bekaert, Filip Pattyn, Bram De Wilde, Paul Coucke, Jo Vandesompele, Kathleen Claes, Jan Hellemans
Primary Institution: Center for Medical Genetics, Ghent University, Ghent, Belgium
Hypothesis
Can massively parallel sequencing (MPS) technologies improve molecular diagnostics for multigenic disorders?
Conclusion
The study provides practical strategies for using MPS in molecular diagnostics, highlighting the required coverage for reliable results.
Supporting Evidence
- The study outlines different PCR amplification strategies for screening multiple genes.
- Over 200 patient samples were evaluated across 10 sequencing runs.
- Guidelines for minimum required coverage were provided based on empirical data.
Takeaway
This study shows how scientists can use new DNA sequencing technology to test many patients at once, making it easier to find genetic problems.
Methodology
The study evaluated 10 GS-FLX sequencing runs on over 200 patient samples using various PCR amplification strategies.
Potential Biases
Potential biases may arise from sample preparation and sequencing processes.
Limitations
The study does not provide a single value for the required minimum coverage due to inter-lab variation.
Participant Demographics
The study involved over 200 patients with various genetic disorders.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website