Quantitative PCR analysis of blood- and saliva-specific microRNA markers following solid-phase DNA extraction

Omelia, Emma J.; Uchimoto, Mari L.; Williams, Graham

doi:10.1016/j.ab.2012.12.024

Cited by 50 publications

(38 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…hsa-miR-205 has been recognised as a marker for saliva by previous authors [5][6][7] and this was supported in the majority of initial screening samples within this investigation. Within the 31 day study expression throughout the menstrual cycle fluctuated over the course of sampling and in some individuals was not expressed at all ( figure 1C).…”

Section: Resultssupporting

confidence: 75%

Characterising the fluctuation of microRNA expression throughout a full menstrual cycle

Bexon

Williams

2015

Forensic Science International: Genetics Supplement Series

Self Cite

View full text Add to dashboard Cite

When processing a crime scene, obtaining a DNA profile that can identify an individual is extremely important. However, the identification of the body fluid that the sample was obtained from could provide key information of the events that occurred. microRNA (miRNA) expression analysis is a technique that has the potential to differentiate body fluids. The presence and expression of body fluid specific miRNA would provide a fast and effective tool for progressing crime scene investigation, especially alleged sexual assault cases. Forensic case work lacks methods for identifying vaginal material, venous blood, menstrual blood and aspermatozoic seminal fluid within samples. A large screening study followed by a 31 day study on five female volunteers was performed utilising RT-qPCR on a large panel of body fluid markers. Screening showed a selection of markers were suitable to differentiate each body fluid, in some cases however, expression fluctuated when analysed over a 31 day period. The data shows that hsa-miR-412 may be suitable for identifying menstrual blood, expression from markers hsa-miR-124 and hsa-miR-205 varied significantly over the 31 day period and between individuals and therefore were less suitable for body fluid identification. The data supports the use of miRNA markers for the identification of certain body fluids such as menstrual and venous blood; however markers for the identification of body fluids such as vaginal material and saliva require further investigation. Keywords:microRNA, Body fluid identification, qPCR, Vaginal material, Menstrual cycle IntroductionThe identification of body fluids is extremely important for progressing forensic case work. In cases of sexual assault, the identification of a body fluid can impact case level and reveal information that can support the prosecution or the defense. The capability of body fluid identification however, is limited. This is due to the non-existence of presumptive identification tests for vaginal material and aspermatozoic seminal fluid. In cases of alleged sexual assault where DNA from a victim is found on a penile swab, the identification of the body fluid present will alter case level; if a technique can differentiate between saliva and vaginal material it may support either the defense or prosecution version of events. In cases of assault where a male cannot produce semen, a DNA profile would be unlikely. If body fluid identification techniques can identify seminal fluid regardless of spermatozoa being present it may give a better understanding of the events of the case. Equally, although presumptive tests for blood are available, there are no currently used methods for differentiating between menstrual and venous blood. In cases of sexual assault, trauma blood is likely to be present, if this can be determined to be venous and not menstrual blood the context of the case would become clearer. The characterization of miRNA: short, stable, non-coding RNA's which modulate gene expression post transcription present a solution to bo...

show abstract

Section: Resultssupporting

confidence: 75%

Characterising the fluctuation of microRNA expression throughout a full menstrual cycle

Bexon

Williams

2015

Forensic Science International: Genetics Supplement Series

Self Cite

View full text Add to dashboard Cite

show abstract

“…Studies have demonstrated great possibilities of saliva to monitor bodily changes, such as metabolic disorders (Gonnissen et al, 2012), anemia (Jagannathan et al, 2012), DNA markers (Omelia et al, 2013), doping control (Thieme, 2012), and especially the physiological changes in athletes resulting from exercise training (Diaz et al, 2013;Moreira et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Analysis of saliva by Fourier transform infrared spectroscopy for diagnosis of physiological stress in athletes

2015

View full text Add to dashboard Cite

Introduction: Saliva is the most promising biofluid to monitor the physiological state of athletes, because this method is not invasive and has low contamination risks. The characterization of saliva by Fourier transform infrared spectroscopy (FT-IR) has been studied as an alternative technique to the standard clinical analysis. However, methodological procedures for saliva analysis are not completely clear, especially in terms of influence of storage conditions and sample preparations for infrared analysis. Thawed saliva includes a precipitate, which may influence the infrared spectral analysis. Thus, the purpose of this study was to show the spectral differences of the precipitate, supernatant, and a combo, as well as the best way to classify the physiological state of the athletes by FT-IR. Methods: The saliva collection was performed before, immediately after, and two hours after a handball match. After the storage of samples at -20 ○ C, it was possible to identify two phases (precipitate and supernatant) and to determine the biochemical differences between the spectra of each phase, which were distinctly analyzed by the second derivative and deconvolution bands. Results: The precipitate and supernatant results showed characteristic bands, especially in the protein regions. All FT-IR spectra were also statistically classified by linear discriminant analysis (LDA), using principal component analysis (PCA). The LDA precipitate and supernatant had lower value when compared to combo spectra (Combination of precipitate and supernatant) with 82%, showing that this combination is the best way to discriminate spectra of saliva samples collected before, immediately after, and 2 h after physical effort. Discussion: The results showed that it is possible to differentiate biochemically the two salivary phases, as well as the importance of the homogenization process of saliva samples to classify the physiological status of athletes using FT-IR.

show abstract

“…Poly-A tail extension [9] was carried out using the miScript II Reverse Transcription kit, (Qiagen, UK) following the manufacturer's instructions, along with the HiSpec Buffer using a Veriti Thermocycler (Life Technologies, UK). Following cDNA synthesis, the product was diluted 1 in 10.…”

Section: Cdna Synthesis Via Poly-a Tail Extensionmentioning

confidence: 99%

“…There are currently two widely used techniques available for the production of larger miRNA amplicons; stem-loop reverse transcription [8] and poly-A tail extension [9]. Both have been utilised for body fluid identification purposes.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Stem-Loop Reverse Transcription and Poly-A Tail Extension in MicroRNA Analysis of Body Fluids

Dunnett¹,

Meer²,

Williams

2015

MIRNA

View full text Add to dashboard Cite

Quantitative PCR analysis of blood- and saliva-specific microRNA markers following solid-phase DNA extraction

Cited by 50 publications

References 12 publications

Characterising the fluctuation of microRNA expression throughout a full menstrual cycle

Characterising the fluctuation of microRNA expression throughout a full menstrual cycle

Analysis of saliva by Fourier transform infrared spectroscopy for diagnosis of physiological stress in athletes

Evaluation of Stem-Loop Reverse Transcription and Poly-A Tail Extension in MicroRNA Analysis of Body Fluids

Contact Info

Product

Resources

About