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. 2022 May 30;13(6):983.
doi: 10.3390/genes13060983.

Evaluation of the Effects of Different Sample Collection Strategies on DNA/RNA Co-Analysis of Forensic Stains

Daniela Lacerenza  1 , Giorgio Caudullo  1 , Elena Chierto  1 , Serena Aneli  1 , Giancarlo Di Vella  1   2 , Marco Barberis  3 , Samuele Voyron  4 , Paola Berchialla  5 , Carlo Robino  1   2
Affiliations

Evaluation of the Effects of Different Sample Collection Strategies on DNA/RNA Co-Analysis of Forensic Stains

Daniela Lacerenza et al. Genes (Basel). .

Abstract

The aim of this study was to evaluate the impact of different moistening agents (RNase-free water, absolute anhydrous ethanol, RNAlater®) applied to collection swabs on DNA/RNA retrieval and integrity for capillary electrophoresis applications (STR typing, cell type identification by mRNA profiling). Analyses were conducted on whole blood, luminol-treated diluted blood, saliva, semen, and mock skin stains. The effects of swab storage temperature and the time interval between sample collection and DNA/RNA extraction were also investigated. Water provided significantly higher DNA yields than ethanol in whole blood and semen samples, while ethanol and RNAlater® significantly outperformed water in skin samples, with full STR profiles obtained from over 98% of the skin samples collected with either ethanol or RNAlater®, compared to 71% of those collected with water. A significant difference in mRNA profiling success rates was observed in whole blood samples between swabs treated with either ethanol or RNAlater® (100%) and water (37.5%). Longer swab storage times before processing significantly affected mRNA profiling in saliva stains, with the success rate decreasing from 91.7% after 1 day of storage to 25% after 7 days. These results may contribute to the future development of optimal procedures for the collection of different types of biological traces.

Keywords: DNA/RNA co-extraction; STR typing; mRNA profiling; swab; trace collection.

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Conflict of interest statement

The authors have declared no conflict of interest.

Figures

Figure 1
Figure 1
Coefficients on the y-axis (log scale) represent the mean increase or decrease in genomic DNA (a) and total RNA (b) concentrations when different moistening agents, storage temperatures (FR: frozen), and storage times before processing were applied to collection swabs, compared to the standard procedure (i.e., swabs moistened with water and stored at room temperature for 7 days before extraction). Significant p-values are marked with an asterisk. LTDB: luminol-treated diluted blood.
Figure 2
Figure 2
Coefficients on the y-axis (log scale) represent the mean increase or decrease in the average peak height (measured in rfu) of the tissue-specific markers (a) and the housekeeping genes (b) when different moistening agents, storage temperatures (FR: frozen), and storage times before processing were applied to collection swabs, compared to the standard procedure (i.e., swabs moistened with water and stored at room temperature for 7 days before extraction). Significant p-values are marked with an asterisk. LTDB: luminol-treated diluted blood.
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