Abstract
Tuberous sclerosis complex (TSC) is an autosomal dominant neurocutaneous syndrome caused by mutations in TSC1 and TSC2. However, 10–15% TSC patients have no mutation identified with conventional molecular diagnostic studies. We used the ultra-deep pyrosequencing technique of 454 Sequencing to search for mosaicism in 38 TSC patients who had no TSC1 or TSC2 mutation identified by conventional methods. Two TSC2 mutations were identified, each at 5.3% read frequency in different patients, consistent with mosaicism. Both mosaic mutations were confirmed by several methods. Five of 38 samples were found to have heterozygous non-mosaic mutations, which had been missed in earlier analyses. Several other possible low-frequency mosaic mutations were identified by deep sequencing, but were discarded as artifacts by secondary studies. The low frequency of detection of mosaic mutations, two (6%) of 33, suggests that the majority of TSC patients who have no mutation identified are not due to mosaicism, but rather other causes, which remain to be determined. These findings indicate the ability of deep sequencing, coupled with secondary confirmatory analyses, to detect low-frequency mosaic mutations.
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Acknowledgments
We thank the TSC patients and families who participated in this study. We thank Paul Au and Hope Northup for the gift of TSC patient DNA. We also thank Edward Szekeres for assistance with AVA software and Michael Egholm of 454 Life Sciences for his support. Supported by NIH NINDS R01 2R37NS031535, and the Tuberous Sclerosis Alliance.
Conflict of interest statement
BET and PB are employees of 454 Life Sciences. The remaining authors have no conflict of interest with regard to this work.
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Qin, W., Kozlowski, P., Taillon, B.E. et al. Ultra deep sequencing detects a low rate of mosaic mutations in tuberous sclerosis complex. Hum Genet 127, 573–582 (2010). https://doi.org/10.1007/s00439-010-0801-z
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DOI: https://doi.org/10.1007/s00439-010-0801-z