scholarly journals mRNA Profiling for Body Fluid Identification by Multiplex Quantitative RT-PCR

2007 ◽  
Vol 0 (0) ◽  
pp. 070917231752009-??? ◽  
Author(s):  
Jane Juusola ◽  
Jack Ballantyne
Keyword(s):  
Body Fluid ◽  
Rt Pcr ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Mrna Profiling

Genome-wide mRNA profiling and multiplex quantitative RT-PCR for forensic body fluid identification

2013 ◽  
Vol 7 (1) ◽  
pp. 143-150 ◽  
Author(s):  
Seong-Min Park ◽  
Seong-Yeon Park ◽  
Jeong-Hwan Kim ◽  
Tae-Wook Kang ◽  
Jong-Lyul Park ◽  
...  
Keyword(s):  
Body Fluid ◽  
Rt Pcr ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Mrna Profiling ◽  
Genome Wide

mRNA profiling for body fluid identification

2008 ◽  
Vol 1 (1) ◽  
pp. 37-38 ◽  
Author(s):  
C. Haas ◽  
B. Klesser ◽  
A. Kratzer ◽  
W. Bär
Keyword(s):  
Body Fluid ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Mrna Profiling

Evaluation of the inclusion of circular RNAs in mRNA profiling in forensic body fluid identification

2017 ◽  
Vol 132 (1) ◽  
pp. 43-52 ◽  
Author(s):  
Yaqi Zhang ◽  
Baonian Liu ◽  
Chengchen Shao ◽  
Hongmei Xu ◽  
Aimin Xue ◽  
...  
Keyword(s):  
Body Fluid ◽  
Circular Rnas ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Mrna Profiling

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

2014 ◽  
Author(s):  
Patrick Danaher ◽  
Robin Lynn White ◽  
Erin Hanson ◽  
Jack Ballantyne
Keyword(s):  
Gene Expression ◽  
Body Fluid ◽  
Body Fluids ◽  
Menstrual Blood ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Mrna Profiling ◽  
Mrna Targets ◽  
Vaginal Secretions ◽  
Specific Mrna

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 tissue/body fluid specific mRNA targets. The body fluids/tissues targeted were peripheral blood, semen, saliva, vaginal secretions, menstrual blood and skin. We tested and compared a simple 5 minute room temperature cellular lysis protocol against standard RNA isolation from same source material as a means to facilitate ease-of-use 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 blood, semen, vaginal secretions, menstrual blood and skin samples 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. As a preliminary indication of the ability of the method to discern admixtures of body fluids, five mixtures were prepared. Two of the five mixtures were called perfectly using the assay algorithm, and one of the component fluids was identified in the each of the 'false negative' mixtures. Crucially, our algorithm produced zero false positive fluid identifications across this study's 98 samples. 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.

Evaluation of one-step RT-PCR multiplex assay for body fluid identification

2021 ◽  
Author(s):  
Qinrui Yang ◽  
Baonian Liu ◽  
Yuxiang Zhou ◽  
Yining Yao ◽  
Zhihan Zhou ◽  
...  
Keyword(s):  
Body Fluid ◽  
Multiplex Assay ◽  
Rt Pcr ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Pcr Multiplex ◽  
One Step

scholarly journals MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation

2010 ◽  
Vol 124 (3) ◽  
pp. 217-226 ◽  
Author(s):  
Dmitry Zubakov ◽  
Anton W. M. Boersma ◽  
Ying Choi ◽  
Patricia F. van Kuijk ◽  
Erik A. C. Wiemer ◽  
...  
Keyword(s):  
Body Fluid ◽  
Rt Pcr ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Microarray Screening

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

F1000Research ◽  
2013 ◽  
Vol 2 ◽  
pp. 281 ◽  
Author(s):  
Erin K. Hanson ◽  
Jack Ballantyne
Keyword(s):  
High Resolution ◽  
Body Fluid ◽  
Biological Fluids ◽  
Fluorescent Dye ◽  
High Resolution Melt ◽  
Gene Target ◽  
Body Fluid Identification ◽  
Fluid Identification ◽  
Mrna Profiling ◽  
Qrt Pcr

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.

Sign in / Sign up

Export Citation Format

Share Document