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Towards Commercialization: Preliminary Developmental Validation of a High Resolution Melt Curve Mixture Prediction Assay and SVM Tool

NCJ Number
Tracey Dawson Green; Andrea Publow
Date Published
March 2023
23 pages

This final technical summary describes the authors’ efforts to more fully develop and assess the value of an emerging laboratory method aimed at de-risking and making more broadly applicable the qPCR platform, which is commonly utilized in forensic DNA laboratories.


The authors report on their research project aimed at developing and assessing the value of a new assay and prediction tool used in forensic DNA laboratories. The project goals were: to test and evaluate the developed integrated mixture screening assay on the QuantStudio™ qPCR platform, which is more consistent with instrumentation used in forensic practice; to complete specific preliminary developmental validation studies to supplement the previously obtained proof-of-concept data; to develop an easy-to-use free, online tool for mixture prediction analysis; and to test the developed integrated mixture screening assay with an analysis tool in a partnering, public forensic laboratory to assure direct applicability to lab practice. The authors’ approach resulted in only a negligible increase in reaction setup time and reaction costs, while providing key sample information for the bench scientist and investigative information for the investigating officers much more quickly. As a result, they suggest that the assay will contribute to saving time and resources that are currently consumed in the lengthy touch DNA and mixture detection, analysis, and reanalysis process. Additionally, the authors suggest that since this tool gives information into the probative, and potentially exclusionary, nature of a sample; it may also help triage which samples to move forward for STR analysis, and minimize additional downstream work.