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                            Library & Multimedia
 Towards developing forensically relevant single-cell pipelines by incorporating direct-to-PCR extraction: compatibility, signal quality, and allele detection
NCJ Number
              304153
Journal

*International Journal of Legal Medicine* Volume: 135 Dated: 2021 Pages: 727–738
Author(s)
          Nidhi Sheth;
                        et al
Date Published

2021
Length
              12 pages
Annotation
              This study demonstrated the feasibility of implementing single-cell pipelines into the forensic process by exploring four metrics of electropherogram (EPG) signal quality; i.e., allele detection rates, peak heights, peak height ratios, and peak height balance across low- to high-molecular-weight short tandem repeat (STR) markers—obtained with four direct-to-PCR extraction treatments and a common post-PCR laboratory procedure.
Abstract
              Current analysis of forensic DNA stains relies on the probabilistic interpretation of bulk-processed samples that represent mixed profiles consisting of an unknown number of potentially partial representations of each contributor. Single-cell methods, in contrast, offer a solution to the forensic DNA mixture problem by incorporating a step that separates cells before extraction. A forensically relevant single-cell pipeline relies on efficient direct-to-PCR extractions that are compatible with standard downstream forensic reagents. In the current research, each treatment was used to extract DNA from 102 single buccal cells; whereupon the amplification reagents were immediately added to the tube and the DNA was amplified/injected using post-PCR conditions known to elicit a limit of detection (LoD) of one DNA molecule. The results show that most cells, regardless of extraction treatment, rendered EPGs with at least a 50-percent true positive allele detection rate and that allele drop-out was not cell independent. Statistical tests demonstrated that extraction treatments significantly impacted all metrics of EPG quality, where the Arcturus® PicoPureTM extraction method resulted in the lowest median allele drop-out rate, highest median average peak height, highest median average peak height ratio, and least negative median values of EPG sloping for GlobalFilerTM STR loci amplified at half volume. The project demonstrated the feasibility of implementing single-cell pipelines for casework purposes and that inferential systems assuming cell independence will not be appropriate in the probabilistic interpretation of a collection of single-cell EPGs. (publisher abstract modified)
      
  
          Additional Details
Grant Number(s)
        2014-DN-BX-0001


  
  
        
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        Computational methods for the interpretation of forensic DNA samples
    
  
  
Date Published: January 1, 2021
    
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