A method to estimate the age of bloodstains using quantitative PCR

Fu, Jun; Allen, Robert W.

doi:10.1016/j.fsigen.2018.12.004

Cited by 29 publications

(36 citation statements)

References 19 publications

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“…Alternatively, in depth analysis of individual transcripts, as recently proposed by Fu et al, might be used to estimate the age of a stain [58]. They reported that in dried bloodstains the 5'-ends of selected mRNA transcripts degraded faster than the 3'-ends.…”

Section: J O U R N a L P R E -P R O O Fmentioning

confidence: 99%

Degradation of human mRNA transcripts over time as an indicator of the time since deposition (TsD) in biological crime scene traces

Salzmann

Russo

Kreutzer

et al. 2021

Forensic Science International: Genetics

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Section: J O U R N a L P R E -P R O O Fmentioning

confidence: 99%

Degradation of human mRNA transcripts over time as an indicator of the time since deposition (TsD) in biological crime scene traces

Salzmann

Russo

Kreutzer

et al. 2021

Forensic Science International: Genetics

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“…Although trends in total RNA concentration tend to follow those found using mRNA transcripts, the latter is much more sensitive as its targeting a specific gene and can determine actual copy numbers [3,14,39]. Most studies use RT-qPCR to amplify the amount of mRNA transcripts .…”

Section: Discussionmentioning

confidence: 99%

“…Not only can RNA identify body fluids, its degradation in forensically relevant stains, such as blood, may also correlate with time [44]. Fu et al [14] found that, when analyzing blood specific mRNA transcripts with qPCR, TSD for bloodstains less than 6 months old could be estimated to within a period of 2 to 4 weeks, while bloodstains aged for 6 months to 1 year old could be estimated 6 to within a period of 4 to 6 weeks. Further, Heneghan et al [39] expanded on the aforementioned study and examined the degradation of three blood-specific transcripts under nine different environmental conditions.…”

Section: Introductionmentioning

confidence: 99%

“…The copyright holder for this preprint (which this version posted April 28, 2021. ; https://doi.org/10.1101/2021.04.28.441847 doi: bioRxiv preprint 4 useful in their own ways, none of these tools help with determining the exact timing of the event, specifically when the crime might have occurred. Previous studies have developed chemometric [2,[9][10][11] and genetic methodologies [3,[12][13][14] that estimate the TSD of bloodstains in controlled laboratory conditions. To this date, these methods have not been accepted for field use, as they largely exhibit insufficient sensitivity and specificity, and high variability between samples and crime scene conditions [3,[9][10][11][12][13]15].…”

Section: Introductionmentioning

confidence: 99%

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Using total RNA quality metrics for time since deposition estimates in degrading bloodstains

Elliott

Stotesbury

Shafer

2021

Preprint

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Determining the age, or time since deposition (TSD), of bloodstains would provide forensic scientists with critical information regarding the timeline of the events of bloodshed during a crime. The physicochemical changes that occur to major biomolecules as a bloodstain dries can be used to estimate the TSD of bloodstains. For example, high-resolution automated gel electrophoresis can be used to quantify the timewise degradation of DNA present in bloodstains. Our study aims to analyze and quantify the timewise degradation trends found in total RNA from bloodstains, expanding the scope of the TSD research which has previously explored DNA and targeted mRNA molecules. Fifty bloodstains were stored in plastic microcentrifuge tubes at room temperature and tested over 10 different timepoints spanning one week. A total of eight RNA metrics were visually assessed and quantified using linear regression. RNA Integrity Number equivalent (RINe), total RNA concentration, and 28S/18S rRNA peak area ratios were retained for further analyses based on their relationship with time and limited correlations. RINe and total RNA concentration both exhibited negative trends over time, highlighting a decrease in quality and quantity. RINe was the RNA metric that demonstrated the greatest association with time (R2 = 0.696). Generalized linear mixed-effects models including donor (biological replicate) as a random effect increased the fit for all RNA metrics to varying degrees, but no significant differences were found between biological replicates for the RINe metric. Our results illustrated the presence of a significant decrease in the retained RNA metrics after 24 hours, suggesting that this method could be used to reliably differentiate day-old bloodstains from older bloodstains. Future work should focus on recreating this study in different environmental conditions, including testing on a variety of substrates.

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“…Also, secondary structures seemed not no influence the persistence rate. They also observed that in dried stains the 5′ end of an mRNA transcript degrades faster than the 3′ end [94]. This differential degradation pattern can be followed with a qPCR assay that quantifies ∼90 bp amplicons produced from the 5′ and 3′ ends of 4 transcripts chosen from the transcriptome of blood.…”

Section: Assigning Body Fluids To Donors In Mixed Body Fluid Stainsmentioning

confidence: 98%

Forensic transcriptome analysis using massively parallel sequencing

Haas

Neubauer

Salzmann

et al. 2021

Forensic Science International: Genetics

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A method to estimate the age of bloodstains using quantitative PCR

Cited by 29 publications

References 19 publications

Degradation of human mRNA transcripts over time as an indicator of the time since deposition (TsD) in biological crime scene traces

Degradation of human mRNA transcripts over time as an indicator of the time since deposition (TsD) in biological crime scene traces

Using total RNA quality metrics for time since deposition estimates in degrading bloodstains

Forensic transcriptome analysis using massively parallel sequencing

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