Next-generation sequencing for disorders of low and high bone mineral density.

Osteoporos Int 2013 Aug 27;24(8):2253-9. Epub 2013 Feb 27.

Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, R814, MS225, Houston, TX 77030, USA.

Unlabelled: To achieve an efficient molecular diagnosis of osteogenesis imperfecta (OI), Ehlers-Danlos syndrome (EDS), and osteopetrosis (OPT), we designed a next-generation sequencing (NGS) platform to sequence 34 genes. We validated this platform on known cases and have successfully identified the causative mutation in most patients without a prior molecular diagnosis.

Introduction: Osteogenesis imperfecta, Ehlers-Danlos syndrome, and osteopetrosis are collectively common inherited skeletal diseases. Evaluation of subjects with these conditions often includes molecular testing which has important counseling and therapeutic and sometimes legal implications. Since several different genes have been implicated in these conditions, Sanger sequencing of each gene can be a prohibitively expensive and time-consuming way to reach a molecular diagnosis.

Methods: In order to circumvent these problems, we have designed and tested a NGS platform that would allow simultaneous sequencing on a single diagnostic platform of different genes implicated in OI, OPT, EDS, and other inherited conditions, leading to low or high bone mineral density. We used a liquid-phase probe library that captures 602 exons (~100 kb) of 34 selected genes and have applied it to test clinical samples from patients with bone disorders.

Results: NGS of the captured exons by Illumina HiSeq 2000 resulted in an average coverage of over 900X. The platform was successfully validated by identifying mutations in six patients with known mutations. Moreover, in four patients with OI or OPT without a prior molecular diagnosis, the assay was able to detect the causative mutations.

Conclusions: In conclusion, our NGS panel provides a fast and accurate method to arrive at a molecular diagnosis in most patients with inherited high or low bone mineral density disorders.

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http://dx.doi.org/10.1007/s00198-013-2290-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3709009PMC
August 2013
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References

(Supplied by CrossRef)

D Krakow et al.
Genet Med 2010

O Makitie et al.
Endocr Dev 2011

ML Warman et al.
Am J Med Genet A 2011

AH Graungaard et al.
Child Care Health Dev 2007

FS Dijk van et al.
Eur J Hum Genet 2012

CS Ku et al.
Ann Neurol 2012

J Wang et al.
2012

X Tian et al.
2012

XY Asan et al.
Genome Biol 2011

EP Homan et al.
J Bone Miner Res 2011

D Baldridge et al.
Hum Mutat 2008

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