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Title

P143 – Comparison of mRNA/miRNA-based Body Fluid Identification with Capillary Electrophoresis (CE) and Massive Parallel Sequencing (MPS)

15:55

Wednesday August 19^th

Station 15

Duration: 12 minutes

04. Forensic biology

Emelie Hanf

In addition to established DNA analysis of forensic traces, the identification of different body fluids and tissues using mRNA analysis was shown as an effective tool. In recent years, a combined mRNA/miRNA multiplex assay for capillary electrophoresis has been developed, which is also suitable for analyzing highly degraded samples. mRNAs exhibit high specificity for particular body fluids. However, due to their long sequences, they are susceptible to degradation. In contrast, miRNAs are short and therefore more stable, but they are less specific for a body fluid.

Therefore, simultaneous analysis of mRNAs and miRNAs provides complementary advantages.

The limitations of this method are the limited number of analyzable markers and the typically low RNA concentration in forensic samples.

On this background it was investigated whether the analysis of body fluids with massive parallel sequencing (MPS) is also possible for low concentrated and degraded samples. First, the capillary electrophoresis (CE) assay was adjusted for usage on MPS through analyzing single-source tissue samples. The biggest challenge was the low length of miRNAs, that would be removed by purification in conventional MPS workflow. After adjusting the purification, the analysis of single-source tissue samples with MPS showed reliable detection of the RNA markers. On basis of these results, the mRNA/miRNA assay was applied to casework samples and samples of a storage study to examine the assay on realistic, low concentrated, and degraded samples.

A comparison between the results of MPS workflow and the established CE workflow showed differences in sensitivity and high differences in specificity. In addition, MPS data showed clearer and error-prone results than the CE data.

Authors

Emelie Hanf-Bähr (Institute of Legal Medicine, Germany)
Sebastian Kunz (Institute of Legal Medicine, Germany)
Malte Bamberg (Institute of Legal Medicine, Germany)

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P143 – Comparison of mRNA/miRNA-based Body Fluid Identification with Capillary Electrophoresis (CE) and Massive Parallel Sequencing (MPS)

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