The Effect of Sample and Sample Matrix on DNA Processing: Mechanisms for the Detection and Management of Inhibition in Forensic Samples

The presence of inhibitory substances in biological forensic samples has, and continues to affect the quality of the data generated following DNA typing processes. Although the chemistries used during the procedures have been enhanced to mitigate the effects of these deleterious compounds, some challenges remain. Inhibitors can be components of the samples, the substrate where samples were deposited or chemical(s) associated to the DNA purification step. Therefore, a thorough understanding of the extraction processes and their ability to handle the various types of inhibitory substances can help define the best analytical processing for any given sample. A series of experiments were conducted to establish the inhibition tolerance of quantification and amplification kits using common inhibitory substances in order to determine if current laboratory practices are optimal for identifying potential problems associated with inhibition. DART mass spectrometry was used to determine the amount of inhibitor carryover after sample purification, its correlation to the initial inhibitor input in the sample and the overall effect in the results. Finally, a novel alternative at gathering investigative leads from samples that would otherwise be ineffective for DNA typing due to the large amounts of inhibitory substances and/or environmental degradation was tested. This included generating data associated with microbial peak signatures to identify locations of clandestine human graves. Results demonstrate that the current methods for assessing inhibition are not necessarily accurate, as samples that appear inhibited in the quantification process can yield full DNA profiles, while those that do not indicate inhibition may suffer from lowered amplification efficiency or PCR artifacts. The extraction methods tested were able to remove >90% of the inhibitors from all samples with the exception of phenol, which was present in variable amounts whenever the organic extraction approach was utilized. Although the results attained suggested that most inhibitors produce minimal effect on downstream applications, analysts should practice caution when selecting the best extraction method for particular samples, as casework DNA samples are often present in small quantities and can contain an overwhelming amount of inhibitory substances.

Criminal Case Processing of Sexual Assaults in Alaska

Discretion plays a role in nearly every facet of the American criminal justice system. It is widely regarded as necessary to do justice but is not without criticisms – especially when it leads to unfavorable or disparate treatment. The role of discretion in sexual assault cases has been particularly scrutinized. Since the majority of sexual assaults do not fit stereotypic beliefs about what constitutes a “real rape” and “genuine victim,” criminal justice officials use their discretion to filter these cases out of the justice system. This study explored this issue by examining two stages of the criminal justice process: the police decision to refer cases for prosecution and the prosecutorial decision to accept referred cases. In doing so, it contributes to this body of literature in three ways. First, it included sexual assault cases that involve Alaska Native victims and suspects. Second, it addressed a gap in the theoretical scholarship by examining the downstream nature of police decision-making. And finally, it examined the formal reasons prosecutors give for charge dispositions. This study found a significant amount of attrition of sexual assault cases as they progressed through the criminal justice system. Moreover, a combination of legally relevant and extralegal factors was found to be important, but not consistently across all types of sexual assaults. Among legal factors, the number of victim injuries was the most consistent predictor. Among extralegal factors, cases that involved Alaska Native suspects had significantly higher odds of case referral and case acceptance compared to white suspects. The effect of suspect race was particularly pronounced in cases with a white victim. Additionally, the findings suggest that not only are Native American defendants more likely to have their cases referred by police, but once referred, they are also more likely to have them accepted for prosecution. Contrary to expectations, victim-suspect relationship, specifically non-stranger assaults, increased the odds of police referral compared to stranger cases. However, the opposite appears to be true for the decision to prosecute cases. Once referred, prosecutors were five times more likely to accept sexual assaults perpetrated by strangers. The implications of these findings are also discussed.

The effect of sample and sample matrix on DNA processing: Mechanisms for the detection and management of inhibition in forensic samples

The presence of inhibitory substances in biological forensic samples has, and continues to affect the quality of the data generated following DNA typing processes. Although the chemistries used during the procedures have been enhanced to mitigate the effects of these deleterious compounds, some challenges remain. Inhibitors can be components of the samples, the substrate where samples were deposited or chemical(s) associated to the DNA purification step. Therefore, a thorough understanding of the extraction processes and their ability to handle the various types of inhibitory substances can help define the best analytical processing for any given sample. A series of experiments were conducted to establish the inhibition tolerance of quantification and amplification kits using common inhibitory substances in order to determine if current laboratory practices are optimal for identifying potential problems associated with inhibition. DART mass spectrometry was used to determine the amount of inhibitor carryover after sample purification, its correlation to the initial inhibitor input in the sample and the overall effect in the results. Finally, a novel alternative at gathering investigative leads from samples that would otherwise be ineffective for DNA typing due to the large amounts of inhibitory substances and/or environmental degradation was tested. This included generating data associated with microbial peak signatures to identify locations of clandestine human graves. Results demonstrate that the current methods for assessing inhibition are not necessarily accurate, as samples that appear inhibited in the quantification process can yield full DNA profiles, while those that do not indicate inhibition may suffer from lowered amplification efficiency or PCR artifacts. The extraction methods tested were able to remove >90% of the inhibitors from all samples with the exception of phenol, which was present in variable amounts whenever the organic extraction approach was utilized. Although the results attained suggested that most inhibitors produce minimal effect on downstream applications, analysts should practice caution when selecting the best extraction method for particular samples, as casework DNA samples are often present in small quantities and can contain an overwhelming amount of inhibitory substances.^