RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains: Results of a fourth and fifth collaborative EDNAP exercise

Haas, Cordula; Hanson, Erin; Anjos, M.J.; Ballantyne, Kaye N.; Banemann, R.; Bhoelai, Bryan; Borges, E.; Carvalho, M.; Courts, Cornelius; Cock, G. De; Drobnič, Katja; Dötsch, M.; Fleming, Rachel; Franchi, Carla Adriene da Silva; Gomes, Iva; Hadzic, Gavrilo; Harbison, SallyAnn; Harteveld, Joyce; Hjort, Benjamin Benn; Hollard, Clémence; Hoff-Olsen, P.; Hüls, Corinna; Keyser, Christine; Maroñas, Olalla; McCallum, Nicola; Moore, David L.; Morling, Niels; Niederstätter, Harald; Noël, Fabrice; Parson, Walther; Phillips, C.; Popielarz, C.; Roeder, Amy D.; Salvaderi, Luca; Sauer, Eva; Schneider, Peter M.; Shanthan, Gnanagowry; Court, Denise Syndercombe; Turanská, M.; Oorschot, Roland A.H. van; Vennemann, Marielle; Vidaki, Athina; Zatkalíková, L.; Ballantyne, Jack

doi:10.1016/j.fsigen.2013.09.009

Cited by 102 publications

(55 citation statements)

References 47 publications

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“…However, further work is required to seek new candidates and include more markers for the identification of vaginal secretion, skin, sweat, tear and urine stains. For example, CYP2B7P1 for vaginal secretion [19], [47] and LCE1C for skin [17], [19], [33] should be considered in future multiplex development. Other more sensitive markers would contribute to the reliable identification of LCN samples in forensic casework.…”

Section: Discussionmentioning

confidence: 99%

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

Xie

Cao

et al. 2014

PLoS ONE

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The identification of human body fluids or tissues through mRNA-based profiling is very useful for forensic investigations. Previous studies have shown mRNA biomarkers are effective to identify the origin of biological samples. In this study, we selected 16 tissue specific biomarkers to evaluate their specificities and sensitivities for human body fluids and tissues identification, including porphobilinogen deaminase (PBGD), hemoglobin beta (HBB) and Glycophorin A (GLY) for circulatory blood, protamine 2 (PRM2) and transglutaminase 4 (TGM4) for semen, mucin 4 (MUC4) and human beta defensin 1(HBD1) for vaginal secretion, matrix metalloproteinases 7 and 11 (MMP7 and MMP11) for menstrual blood, keratin 4(KRT4) for oral mucosa, loricrin (LOR) and cystatin 6 (CST6) for skin, histatin 3(HTN3) for saliva, statherin (STATH) for nasal secretion, dermcidin (DCD) for sweat and uromodulin (UMOD) for urine. The above mentioned ten common forensic body fluids or tissues were used in the evaluation. Based on the evaluation, a reverse transcription (RT) PCR multiplex assay, XCYR1, which includes 12 biomarkers (i.e., HBB, GLY, HTN3, PRM2, KRT4, MMP11, MUC4, DCD, UMOD, MMP7, TGM4, and STATH) and 2 housekeeping genes [i.e., glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 18SrRNA], was developed. This assay was further validated with real casework samples and mock samples (with both single source and mixture) and it was approved that XCYR1 is effective to identify common body fluids or tissues (i.e., circulatory blood, saliva, semen, vaginal secretion, menstrual blood, oral mucosa, nasal secretion, sweat and urine) in forensic casework samples.

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Section: Discussionmentioning

confidence: 99%

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

Xie

Cao

et al. 2014

PLoS ONE

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“…Various multiplex assays and scoring systems (to account for results from multiple markers for one body fluid) have been developed and are utilized on evidential‐type samples (Albani & Fleming, ; Ballantyne & Juusola, ; Fleming & Harbison, ; Haas, Hanson, & Ballantyne, ; Juusola & Ballantyne, ; Lindenbergh et al, ; Roeder & Haas, ; Visser, Zubakov, Ballantyne, & Kayser, ). The European DNA Profiling Group have undertaken various exercises on RNA/DNA coanalysis for body fluid identification, using mRNA multiplexes, and STR profiling on blood, saliva, semen, menstrual blood, and vaginal material (Gill et al, ; Haas et al, ; Haas et al, ). Similarly, the European Forensic Genetics Network of Excellence carried out a collaborative study testing a 20‐marker body fluid and skin typing multiplex across nine laboratories (Van Den Berge et al, ).…”

Section: Body Fluids In Forensic Sciencementioning

confidence: 99%

A review of direct polymerase chain reaction of DNA and RNA for forensic purposes

Lynch

Fleming

2019

WIREs Forensic Science

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Short tandem repeat (STR) profiling is frequently carried out on biological stains in forensic casework, and the application of messenger RNA (mRNA) body fluid identification is becoming more prevalent. Increasing constraints on analysis time, resources, an increase in the number of samples for analysis, and a large backlog of cases creates significant demand for quicker methods that direct polymerase chain reaction (PCR) can help overcome. Direct PCR bypasses DNA and/or RNA extraction, and in some cases, quantification by adding the sample or substrate punch to the amplification reaction. This method is currently used in some laboratories on reference samples. Direct PCR is also a component of portable devices, which could improve efficiency even further by producing results at the scene, and enabling triaging of samples for further testing. Portable devices are currently used for STR profiling of reference samples but not for mRNA profiling. Various concerns regarding the reliability of the method and equipment (portable devices) must first be addressed before incorporation into forensic casework. This review discusses the limitations and factors that need to be considered for direct PCR to be used for the analysis of casework samples, including being used for mRNA body fluid identification. This article is categorized under: Forensic Biology > Forensic DNA Technologies Forensic Biology > Body Fluid Identification Forensic Biology > Applications of RNA

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“…blood -ANK1, ALAS2, HBB; semen -PRM1, PRM2, TGM4, SEMG1; saliva -HTN3; vaginal secretions -CYP2B7P1; menstrual blood -MMP10, MMP7, LEFTY2) [5,8]. These genes, and others, have been extensively evaluated by the forensic community including numerous collaborative EDNAP studies on RNA profiling for body fluid identification [17,18,[37][38][39][40], thus aiding our initial selection of genes for the targeted RNA sequencing assay. We designed and evaluated six targeted oligonucleotide primer pools (TOPs;…”

Section: Candidate Selectionmentioning

confidence: 99%

Messenger RNA biomarker signatures for forensic body fluid identification revealed by targeted RNA sequencing

Hanson

Ingold

Haas

et al. 2018

Preprint

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The recovery of a DNA profile from the perpetrator or victim in criminal investigations can provide valuable ‘source level’ information for investigators. However, a DNA profile does not reveal the circumstances by which biological material was transferred. Some contextual information can be obtained by a determination of the tissue or fluid source of origin of the biological material as it is potentially indicative of some behavioral activity on behalf of the individual that resulted in its transfer from the body. Here, we sought to improve upon established RNA based methods for body fluid identification by developing a targeted multiplexed next generation mRNA sequencing assay comprising a panel of approximately equal sized gene amplicons. The multiplexed biomarker panel includes several highly specific gene targets with the necessary specificity to definitively identify most forensically relevant biological fluids and tissues (blood, semen, saliva, vaginal secretions, menstrual blood and skin). In developing the biomarker panel we evaluated 66 gene targets, with a progressive iteration of testing target combinations that exhibited optimal sensitivity and specificity using a training set of forensically relevant body fluid samples. The current assay comprises 33 targets: 6 blood, 6 semen, 6 saliva, 4 vaginal secretions, 5 menstrual blood and 6 skin markers. We demonstrate the sensitivity and specificity of the assay and the ability to identify body fluids in single source and admixed stains. A 16 sample blind test was carried out by one lab with samples provided by the other participating lab. The blinded lab correctly identified the body fluids present in 15 of the samples with the major component identified in the 16th. Various classification methods are being investigated to permit inference of the body fluid/tissue in dried physiological stains. These include the percentage of reads in a sample that are due to each of the 6 tissues/body fluids tested and inter-sample differential gene expression revealed by agglomerative hierarchical clustering.

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RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains: Results of a fourth and fifth collaborative EDNAP exercise

Cited by 102 publications

References 47 publications

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

Development of Highly Sensitive and Specific mRNA Multiplex System (XCYR1) for Forensic Human Body Fluids and Tissues Identification

A review of direct polymerase chain reaction of DNA and RNA for forensic purposes

Messenger RNA biomarker signatures for forensic body fluid identification revealed by targeted RNA sequencing

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