United States Department of Transportation
i
Postmortem Samples from Aviation Accident Victims Maintain Tissue-Specific mRNA Expression Profiles
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2017-11-01
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[PDF-841.46 KB]
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Abstract:To determine whether an aerospace medical factor such as hypoxia or sleep deprivation played a role in the cause of an aviation accident, gene expression marker panels are in development based on results from live subject studies in humans. The translation of marker panels from the discovery phase to a forensic setting requires that RNA stability and the factors that affect it be assessed in forensic samples. To assess the utility of postmortem blood samples for gene expression analysis, RNA was purified from aviation accident victim blood and tissue samples, and gene expression levels from a panel of genes assessed by qRT-PCR. Live subject data was obtained from publicly available microarray data. Correlation analyses were performed between all victim samples and tissue specific victim samples compared to live subject microarray expression data. Due to observed prokaryotic rRNA in human samples, universal primers for bacterial 16S rRNA were characterized on purified prokaryotic total RNA and victim samples. RNA integrity was variable between victim samples but there was no correlation between RNA integrity and yield or postmortem interval. Expression patterns between victims were generally concordant. Blood, brain, and muscle were correlated between live subjects and victims. The assay for bacterial rRNA was found to be sensitive for very low levels of purified bacterial RNA and highly specific for bacterial rRNA when challenged with mixtures of bacterial and human RNA. In conclusion, victim sample RNAs exhibits a wide range of degradation but qRT-PCR data maintains the relative expression values between target genes. That victim samples had similar expression patterns to live subject data suggests that marker panels developed in human subject studies can be translated to accident investigation and will allow additional information to be incorporated into medical determinations of accident causality
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