Validation of NGS-Based DNA Testing and Implementation of Quality Control Procedures
The rapid adoption of next-generation sequencing (NGS) (also known as massively parallel sequencing; MPS) techniques has revolutionized the way molecular diagnosis is performed in a clinical diagnostic laboratory. Hundreds to thousands of genes can be analyzed simultaneously, and samples can be multiplexed and sequenced in parallel with deep coverage. To bring such a complex technology to clinical diagnostic laboratories, limitations and challenges should be recognized when designing and implementing assays for routine clinical use. While most NGS platforms can generate enormous amount of data in a massively parallel manner, the complexities of test validation and the implementation of quality control procedures have posed tremendous challenges on quality assurance. A quality assurance program is an integral part of clinical laboratory operations, in particular for laboratories that are adopting new technologies to provide state-of-the-art genetic testing services. In this chapter, we describe the steps taken in implementation of test validation and quality control procedures in our clinical molecular diagnostic laboratory.
KeywordsShipping Assure Clarification Retinitis Dideoxy
- 1.Zhang W, Cui H, Wong LJ (2012) Application of next generation sequencing to molecular diagnosis of inherited diseases. Top Curr Chem 2012 May 11 (Epub ahead of print)Google Scholar
- 11.Hu H, Wrogemann K, Kalscheuer V, Tzschach A, Richard H, Haas SA, Menzel C, Bienek M, Froyen G, Raynaud M, Van Bokhoven H, Chelly J, Ropers H, Chen W (2009) Mutation screening in 86 known X-linked mental retardation genes by droplet-based multiplex PCR and massive parallel sequencing. Hugo J 3(1–4):41–49PubMedCrossRefGoogle Scholar
- 13.Voelkerding KV, Dames S, Durtschi JD (2010) Next generation sequencing for clinical diagnostics-principles and application to targeted resequencing for hypertrophic Cardiomyopathy: a paper from the 2009 William Beaumont Hospital symposium on molecular pathology. J Mol Diagn 12(5):539–551PubMedCrossRefGoogle Scholar
- 17.Bell CJ, Dinwiddie DL, Miller NA, Hateley SL, Ganusova EE, Mudge J, Langley RJ, Zhang L, Lee CC, Schilkey FD, Sheth V, Woodward JE, Peckham HE, Schroth GP, Kim RW, Kingsmore SF (2011) Carrier testing for severe childhood recessive diseases by next-generation sequencing. Sci Transl Med 3(65):65ra64Google Scholar
- 20.Chen B, Gagnon M, Shahangian S, Anderson NL, Howerton DA, Boone JD (2009) Good laboratory practices for molecular genetic testing for heritable diseases and conditions. MMWR Recomm Rep 58(RR-6):1–37Google Scholar