Accelerated DNA profiling through workflow reconfiguration: Practical experience from 2-year postmortem skeletal remains in hostage repatriation.

The identification of human remains in mass-casualty incidents requires rapid and reliable forensic DNA analysis, particularly when dealing with degraded skeletal material. Although rapid forensic identification is a fundamental objective in all disaster victim identification (DVI) operations, the repatriation of deceased hostages from the October 7, 2023, attack, c. 2 years postmortem (PM), imposed an additional challenge of a fixed operational deadline dictated by the implementation of the hostages-prisoners exchange agreement. To accelerate STR profiling, we reconfigured the conventional sequential process (extraction, quantification, amplification) into a flexible, decision-driven workflow. The new workflow incorporates a shortened 1.5-h DNA incubation, predefined dual-volume PCR amplification, and deferred DNA quantification. Built-in contingency mechanisms including multi-sample per case and enhanced 33-Cycle PCR-enabled immediate progression to alternative pathways when required. The accelerated workflow was applied in a real-world operational setting involving 37 cases (79 samples) collected during multiple repatriation phases. Informative STR profiles were obtained from 90% (71/79) of samples within 4 h per repatriation phase. Although eight individual samples remained non-informative, the integrated contingency mechanisms enabled successful resolution of all 37 cases (100%). Informative profiles were generated across a broad range of DNA quantities, including low-template samples, and from diverse skeletal elements. By comparison, the RapidHIT™ system generated an informative profile in only one of 21 samples. These findings demonstrate that restructuring the DNA workflow into a flexible, decision-driven process with built-in contingency mechanisms enables rapid and reliable profiling. The proposed workflow provides a practical operational framework for time-critical forensic DNA identification.