Verification of the analytical validity of next-generation sequencing methods

Abstract

To validate the analytical validity of next-generation sequencing methods, it is necessary to examine whether the variants can be correctly detected using genomic DNA with known variant information. The benchmark human genome variants in the human reference genome published by the Genome In A Bottle (GIAB) consortium at the US National Institute of Standards and Technology (NIST) are ideal for verifying analytical validity. Here, we present the methods and results of analytical validation of next-generation sequencing method that we conducted using GIAB data and standard human genome DNA, and demonstrate that analytical validation is possible at facilities in Japan. In addition, to verify the analytical validity of the data as an institution, i.e., whether pathological variants can be correctly detected based on the patient phenotype, we present an example of our use of an external quality assessment (EQA), GenQA in the UK. This study discusses the method for verifying the analytical validity of next-generation sequencing methods.