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Detection of Epithelial Cells in Dried Blood Stains by Reverse Transcriptase-Polymerase Chain Reaction

NCJ Number
183898
Journal
Journal of Forensic Sciences Volume: 44 Issue: 6 Dated: November 1999 Pages: 1232-1236
Author(s)
Martin Bauer M.D.; Alexandra Kraus M.D.; Dieter Patzelt M.D.
Date Published
November 1999
Length
5 pages
Annotation
This study investigated the extraction of RNA suitable for reverse transcriptase-polymerase chain reaction (RT-PCR) from dried blood stains stored for up to 6 months.
Abstract
The identification of menstrual blood stains can be improved by detection of messenger ribonucleic acid (mRNA) specific for epithelial (endometrial) cells. RNA molecules, however, are believed to be unstable and require careful sample processing. With a modified RNA isolation protocol, it was possible to obtain RNA from dried blood stains with at least 5x10 squared leukocytes. In an additional experiment, the authors evaluated the RNA isolation from mixed stains composed of leukocytes and T47D cells, a breast cancer-derived cell line with epithelial origin. Detection of 10 squared T47D cells in a total number of 10-to-the-fifth-power leukocytes was possible by amplification of cytokeratin 19 mRNA and progesterone receptor-mRNA specific for hormonally regulated epithelial cells. In both experiments amplification results were not dependent on storage time with similar data from 1 day to 6 months. Furthermore, it was possible to identify dried menstrual blood samples by showing the presence of mRNA specific for epithelial cells. These results show for the first time that RNA suitable for RT-PCR can be isolated from forensic specimens stored up to at least 6 months, and that a small number of epithelial (endometrial) cells can be identified in dried blood specimens. Using this method, it will be possible to identify the origin of small and partially degraded blood samples that can be especially useful in forensic evaluation of sexual offense cases. 4 figures and 19 references