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                            Library & Multimedia
 A quantifilerTM trio-based HRM screening assay for the accurate prediction of single source versus mixed biological samples
NCJ Number
              307397
Journal

*International Journal of Legal Medicine* Volume: Online Dated: 2023
Author(s)
          Dayanara Torres;
                        Chastyn Smith;
                        Andrea L. Williams;
                        Jordan O. Cox;
                        Sarah J. Seashols-Williams;
                        Edward L. Boone;
                        Tracey Dawson Green
Date Published

2023
Abstract
              At present, the forensic DNA workflow is not capable of providing information about the contributor status (single source vs. multiple contributors) of evidentiary samples prior to end-point analysis. This exacerbates the challenges inherent to mixtures and low-template DNA samples. If additional sample information could be provided earlier in the workflow, protocols could be implemented to mitigate these challenges. An integrated Quantiplex®- high resolution melt (HRM) assay was shown to be effective in distinguishing between single source and mixture DNA samples; however, integration of the HRM assay into a more commonly used chemistry would be beneficial to the practitioner community. Thus, the assay was redesigned as an integrated QuantifilerTM Trio-HRM assay, which included the identification of a new DNA-binding dye, an increased reaction volume, and the establishment of new data analysis and standard curve metrics for all targets. This redesigned assay produced quantification values and qualitative values that were comparable to those produced when the same samples were tested using the standard QuantifilerTM Trio chemistry and settings. Further, STR profiles generated with quantification values produced from the integrated QuantifilerTM Trio-HRM assay and standard QuantifilerTM Trio chemistry were complete and fully concordant. Most importantly, the integrated QuantifilerTM Trio-HRM assay was able to accurately predict whether a sample was single source or a mixture 79.2% of the time, demonstrating the potential of this approach. With the incorporation of an expanded training set for prediction modeling, and completion of critical developmental validation studies, this assay could prove useful to the forensic DNA practitioner community. (Published Abstract Provided)
      
  
          Additional Details
Grant Number(s)
        2019-DU-BX-0003


  
  
        
  Agencies
        NIJ-Sponsored


  
  
        
  Publication Type
        Research (Applied/Empirical)


  
  
        
  Award
        Towards Commercialization: Preliminary developmental validation of a high resolution melt curve mixture prediction assay and SVM tool
    
  
  
Date Published: January 1, 2023
    
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        Forensic biology
        DNA (Deoxyribonucleic Acid)
        Trace DNA



  
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