Efficacy of Several Candidate Protein Biomarkers in the Differentiation of Vaginal from Buccal Epithelial Cells*

Simons, Joanne L.; Vintiner, Sue

doi:10.1111/j.1556-4029.2012.02158.x

Cited by 15 publications

(7 citation statements)

References 25 publications

(26 reference statements)

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“…65,90 Some of these markers have been found to be highly expressed in vaginal secretions. 65,91 Regardless of the technical approach used, reporting guidelines based on laboratory validation are needed. Proposed reporting strategies for end-point PCR approaches vary.…”

Section: Rna-and Dna-based Technologiesmentioning

confidence: 99%

Forensic body fluid identification: state of the art

Harbison¹,

Fleming²

2016

RRFMS

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Body fluid identification is a key component in the forensic scientists' tool box and has been carried out both at the crime scene and in the laboratory for many years. Historically, methods relied on (bio) chemical-based tests, many of which lacked specificity. In this review, current technologies for identifying body fluids are described including the use of RNA (mRNA and miRNA), epigenetics, spectroscopic techniques such as Raman spectroscopy and microspectrophotometry, biosensors, and immunochromatographic methods which are outlined alongside their strengths and weaknesses. The potential for new insights into the identification of cells from new technologies such as massively parallel sequencing is explored.

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Section: Rna-and Dna-based Technologiesmentioning

confidence: 99%

Forensic body fluid identification: state of the art

Harbison¹,

Fleming²

2016

RRFMS

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“…The field of forensic miRNA profiling, although promising, is less mature in terms of there being an international consensus on the identity and specificity of the best body fluid specific miRNA targets. Other non-RNA methods for body fluid identification have been recently investigated including the use of epigenetic [23–29] and proteomic [30–32] biomarkers. Although exhibiting some promise, epigenetic markers have not been identified for all of the important common body fluids and tissues such as vaginal secretions and skin.…”

Section: Introductionmentioning

confidence: 99%

Facile semi-automated forensic body fluid identification by multiplex solution hybridization of NanoString® barcode probes to specific mRNA targets

Danaher¹,

White²,

Hanson³

et al. 2014

Preprint

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Abstract:A DNA profile from the perpetrator does not reveal, per se, the circumstances by which it was transferred. Body fluid identification by mRNA profiling may allow extraction of contextual ‘activity level’ information from forensic samples. Here we describe the development of a prototype multiplex digital gene expression (DGE) method for forensic body fluid/tissue identification based upon solution hybridization of color-coded NanoString® probes to 23 mRNA targets. The method identifies peripheral blood, semen, saliva, vaginal secretions, menstrual blood and skin. We showed that a simple 5 minute room temperature cellular lysis protocol gave equivalent results to standard RNA isolation from the same source material, greatly enhancing the ease-of-use of this method in forensic sample processing. We first describe a model for gene expression in a sample from a single body fluid and then extend that model to mixtures of body fluids. We then describe calculation of maximum likelihood estimates (MLEs) of body fluid quantities in a sample, and we describe the use of likelihood ratios to test for the presence of each body fluid in a sample. Known single source samples of blood, semen, vaginal secretions, menstrual blood and skin all demonstrated the expected tissue-specific gene expression for at least two of the chosen biomarkers. Saliva samples were more problematic, with their previously identified characteristic genes exhibiting poor specificity. Nonetheless the most specific saliva biomarker, HTN3, was expressed at a higher level in saliva than in any of the other tissues. Crucially, our algorithm produced zero false positives across this study’s 89 unique samples. As a preliminary indication of the ability of the method to discern admixtures of body fluids, five mixtures were prepared. The identities of the component fluids were evident from the gene expression profiles of four of the five mixtures. Further optimization of the biomarker ‘CodeSet’ will be required before it can be used in casework, particularly with respect to increasing the signal-to-noise ratio of the saliva biomarkers. With suitable modifications, this simplified protocol with minimal hands on requirement should facilitate routine use of mRNA profiling in casework laboratories.

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“…Thus there remains a need to develop definitive molecular based strategies for the conclusive identification of all forensically relevant biological fluids and tissues. Although protein 1– 3 , epigenetic DNA markers 4– 10 and microRNAs 11– 17 are promising examples of such a molecular based approach, messenger RNA (mRNA) profiling represents the current gold standard in this area due to the identification and development of a number of specific and sensitive mRNA assays for the identification of all of the forensically-relevant body fluids and tissues, namely blood, semen, saliva, vaginal secretions, menstrual blood and skin 13, 18– 36 . Importantly, messenger RNA markers are surprisingly stable in the dried state in that they can be successfully detected in environmentally impacted and partially degraded samples 20, 22, 25, 26, 37– 40 .…”

Section: Introductionmentioning

confidence: 99%

Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis

Hanson¹,

Ballantyne

2014

F1000Res

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Positive identification of the nature of biological material present on evidentiary items can be crucial for understanding the circumstances surrounding a crime. However, traditional protein-based methods do not permit the identification of all body fluids and tissues, and thus molecular based strategies for the conclusive identification of all forensically relevant biological fluids and tissues need to be developed. Messenger RNA (mRNA) profiling is an example of such a molecular-based approach. Current mRNA body fluid identification assays involve capillary electrophoresis (CE) or quantitative RT-PCR (qRT-PCR) platforms, each with its own limitations. Both platforms require the use of expensive fluorescently labeled primers or probes. CE-based assays require separate amplification and detection steps thus increasing the analysis time. For qRT-PCR assays, only 3-4 markers can be included in a single reaction since each requires a different fluorescent dye.To simplify mRNA profiling assays, and reduce the time and cost of analysis, we have developed single- and multiplex body fluid High Resolution Melt (HRM) assays for the identification of common forensically relevant biological fluids and tissues. The incorporated biomarkers include IL19 (vaginal secretions), IL1F7 (skin), ALAS2 (blood), MMP10 (menstrual blood), HTN3 (saliva) and TGM4 (semen). The HRM assays require only unlabeled PCR primers and a single saturating intercalating fluorescent dye (Eva Green). Each body-fluid-specific marker can easily be identified by the presence of a distinct melt peak. Usually, HRM assays are used to detect variants or isoforms for a single gene target. However, we have uniquely developed duplex and triplex HRM assays to permit the simultaneous detection of multiple targets per reaction. Here we describe the development and initial performance evaluation of the developed HRM assays. The results demonstrate the potential use of HRM assays for rapid, and relatively inexpensive, screening of biological evidence.

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Efficacy of Several Candidate Protein Biomarkers in the Differentiation of Vaginal from Buccal Epithelial Cells*

Cited by 15 publications

References 25 publications

Forensic body fluid identification: state of the art

Forensic body fluid identification: state of the art

Facile semi-automated forensic body fluid identification by multiplex solution hybridization of NanoString® barcode probes to specific mRNA targets

Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis

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