CONTEXT: DNA sequencing is the method of choice for mutation detection in many genes. OBJECTIVES: To demonstrate the analytical accuracy and reliability of DNA sequencing assays developed in clinical laboratories. Only general guidelines exist for the validation of these tests. We provide examples of assay validation strategies for DNA sequencing tests. DESIGN: We discuss important design and validation considerations. RESULTS: The validation examples include an accuracy study to evaluate concordance between results obtained by the newly designed assay and analyzed by another method or laboratory. Precision (reproducibility) studies are performed to determine the robustness of the assay. To assess the quality of sequencing assays, several sequence quality measures are available. In addition, assessing the ability of primers to specifically and robustly amplify target regions before sequencing is important. CONCLUSION: Protocols for validation of laboratory-developed sequencing assays may vary between laboratories. An example summary of a validation is provided.
CONTEXT: DNA sequencing is the method of choice for mutation detection in many genes. OBJECTIVES: To demonstrate the analytical accuracy and reliability of DNA sequencing assays developed in clinical laboratories. Only general guidelines exist for the validation of these tests. We provide examples of assay validation strategies for DNA sequencing tests. DESIGN: We discuss important design and validation considerations. RESULTS: The validation examples include an accuracy study to evaluate concordance between results obtained by the newly designed assay and analyzed by another method or laboratory. Precision (reproducibility) studies are performed to determine the robustness of the assay. To assess the quality of sequencing assays, several sequence quality measures are available. In addition, assessing the ability of primers to specifically and robustly amplify target regions before sequencing is important. CONCLUSION: Protocols for validation of laboratory-developed sequencing assays may vary between laboratories. An example summary of a validation is provided.
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Authors: Neal I Lindeman; Philip T Cagle; Mary Beth Beasley; Dhananjay Arun Chitale; Sanja Dacic; Giuseppe Giaccone; Robert Brian Jenkins; David J Kwiatkowski; Juan-Sebastian Saldivar; Jeremy Squire; Erik Thunnissen; Marc Ladanyi Journal: Arch Pathol Lab Med Date: 2013-04-03 Impact factor: 5.534
Authors: Chih-Jian Lih; David J Sims; Robin D Harrington; Eric C Polley; Yingdong Zhao; Michele G Mehaffey; Thomas D Forbes; Biswajit Das; William D Walsh; Vivekananda Datta; Kneshay N Harper; Courtney H Bouk; Lawrence V Rubinstein; Richard M Simon; Barbara A Conley; Alice P Chen; Shivaani Kummar; James H Doroshow; Paul M Williams Journal: J Mol Diagn Date: 2015-11-18 Impact factor: 5.568
Authors: Piotr Rutkowski; Aleksandra Gos; Monika Jurkowska; Tomasz Switaj; Wirginiusz Dziewirski; Marcin Zdzienicki; Konrad Ptaszyński; Wanda Michej; Andrzej Tysarowski; Janusz A Siedlecki Journal: Oncol Lett Date: 2014-05-08 Impact factor: 2.967
Authors: Bruce Budowle; Nancy D Connell; Anna Bielecka-Oder; Rita R Colwell; Cindi R Corbett; Jacqueline Fletcher; Mats Forsman; Dana R Kadavy; Alemka Markotic; Stephen A Morse; Randall S Murch; Antti Sajantila; Sarah E Schmedes; Krista L Ternus; Stephen D Turner; Samuel Minot Journal: Investig Genet Date: 2014-07-30