Multiplex high resolution melt (HRM) messenger RNA profiling assays for body fluid identification |
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| Affiliation: | 1. National Center for Forensic Science, University of Central Florida, Orlando, FL, USA;2. University of Central Florida, Department of Chemistry, Orlando, FL, USA;1. Network of Forensic Science Institutes, Institute of Forensic Medicine, DNA Laboratory, Budapest, Hungary;2. Research Centre for Natural Sciences of the HAS, Department of Complex Systems, Budapest, Hungary;1. Laboratorio de Genética Molecular de Cruz Vital – Cruz Roja Ecuatoriana Quito, Ecuador;2. Laboratorio GENES Ltda, Medellin, Colombia;1. Institute of Legal Medicine, Faculty of Medicine, University of Cologne, Cologne, Germany;2. IPATIMUP – Institute of Molecular Pathology and Immunology of the University of Porto, Porto, Portugal;3. Forensic Genetics Unit, Institute of Legal Medicine, University of Santiago de Compostela, Santiago de Compostela, Spain;1. Forensics Genetic Service, Central Delegation, National Institute of Legal Medicine and Forensic Sciences, I.P., Coimbra, Portugal;2. Cencifor, Forensic Science Centre, Portugal;3. National Institute of Legal Medicine and Forensic Sciences, I.P., Portugal;4. Department of Biology, University of Aveiro, Portugal;1. Armed Forces DNA Identification Laboratory, Armed Forces Medical Examiner System, United States;2. American Registry of Pathology, United States;3. Flinders University, Australia;4. National Institute of Standards and Technology, United States |
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| Abstract: | Traditional body fluid identification methods use a variety of technologically diverse techniques that do not permit the identification of all body fluids. Definitive identification of the biological material present can be crucial to a fuller understanding of the circumstances pertaining to a crime. Thus definitive molecular based strategies for the conclusive identification of forensically relevant biological fluids need to be developed. Messenger (mRNA) profiling is an example of such a molecular based approach.Current mRNA body fluid identification assays typically involve either 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 time required for analysis. For qRT-PCR assays, only 3 or 4 markers can be included in a single reaction since each requires a different fluorescent dye. To simplify mRNA profiling assays and to reduce the time and cost of analysis, we have developed multiplex high resolution melt (HRM) assays that provide an identification of all forensically relevant biological fluids and tissues. |
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| Keywords: | Messenger RNA (mRNA) profiling Body fluid identification High resolution melt (HRM) analysis Forensic science |
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