Inter-tool Analysis of a NIST Dataset for Assessing Baseline Nucleic Acid Sequence Screening

Introduction: Nucleic acid synthesis is a dual-use technology that can benefit fields such as biology, medicine, and information storage. However, synthetic nucleic acids could also potentially be used negligently and ultimately cause harm, or be used with malicious intent to cause harm. Thus, this technology needs to be appropriately safeguarded. Sequence screening is one component of a biosecurity protocol for preventing such harm and consists of identifying Sequences of Concern (SOCs). There exist many fit-for-purpose tools that have been developed for nucleic acid synthesis sequence screening. However, questions remain regarding their performance with respect to the consistency of screening. Methods: To aid in determining if screening tools are harmonized in regard to baseline sequence screening (which represents a minimum acceptable level of performance), the National Institute of Standards and Technology (NIST) constructed a test dataset based on current screening recommendations. NIST then sent blinded datasets to sequence screening tool developers for testing. Results: Overall, there was a general agreement between the tools and NIST labels given to the sequences, and all tools had a baseline performance of >95% sensitivity and >97% accuracy. Disagreement on specific sequences largely arose from single tools and could be traced to differences in defining a SOC and/or methodological differences in screening algorithms.