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

Authors

  • Jane Juusola Ph.D.,

    1. Graduate Program in Biomolecular Sciences, University of Central Florida, Orlando, FL.
    2. Department of Chemistry, University of Central Florida, Orlando, FL.
    3. National Center for Forensic Science, University of Central Florida, Orlando, FL.
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  • Jack Ballantyne Ph.D.

    1. Graduate Program in Biomolecular Sciences, University of Central Florida, Orlando, FL.
    2. Department of Chemistry, University of Central Florida, Orlando, FL.
    3. National Center for Forensic Science, University of Central Florida, Orlando, FL.
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  • *

    Presented in part at SAFS, MAFS, MAAFS, CFSF Joint Meeting in Orlando, FL, September 23, 2004, and Promega’s 16th International Symposium on Human Identification in Grapevine, TX, September 29, 2005.

Additional information and reprint requests:
Jack Ballantyne, Ph.D.
Associate Professor
Department of Chemistry
University of Central Florida
Building 5, 4000 Central Boulevard
Orlando, FL 32816-2366
E-mail: jballant@mail.ucf.edu

Abstract

Abstract:  An alternative approach to conventional protein-based body fluid identification is gene expression profiling analysis. In the present work, we report the development of sensitive and robust multiplex quantitative reverse transcriptase-PCR assays for the identification of blood, saliva, semen, and menstrual blood. Each body fluid assay comprises a triplex system that detects transcripts from two body fluid-specific genes and a housekeeping gene GAPDH. The body fluid-specific genes include erythroid δ-aminolevulinate synthase (ALAS2) and β-spectrin (SPTB) for blood, statherin (STATH) and histatin 3 (HTN3) for saliva, protamine 1 (PRM1) and protamine 2 (PRM2) for semen, and matrix metalloproteinase 7 (MMP7) and matrix metalloproteinase 10 (MMP10) for menstrual blood. Normalization of both body fluid-specific genes to the housekeeping gene by means of appropriate cycle threshold metrics ensures the high specificity of each assay for its cognate body fluid.

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