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mRNA Profiling for Body Fluid Identification by Reverse Transcription Endpoint PCR and Realtime PCR

NCJ Number
226992
Journal
Forensic Science International: Genetics Volume: 3 Issue: 2 Dated: March 2009 Pages: 80-88
Author(s)
C. Haas; B. Klesser; C. Maake; W. Bar; A. Kratzer
Date Published
March 2009
Length
9 pages
Annotation
In this study, a reverse transcription endpoint polymerase chain reaction (PCR) and a realtime PCR method (two mRNA profiling methods) were developed and compared to conventional enzymatic and immunologic tests for the identification of blood, saliva, semen, vaginal secretions, and menstrual blood.
Abstract
Findings show that realtime PCR did not prove to be any more sensitive than endpoint PCR, but both methods were shown to be suitable for the identification of body fluids in forensic stains. Major advantages of mRNA profiling are the possibility of detecting several body fluids in one multiplex reaction and the simultaneous DNA isolation without loss of material. The results show that both endpoint PCR and realtime PCR are suitable for the identification of body fluids found in forensic stains and represent an effective alternative to conventional enzymatic and immunologic tests. Two novel endpoint PCR multiplex assays were created: multiplex 1 for screening the forensically most relevant body fluids and multiplex 2 for the detection of blood, vaginal secretions, and menstrual blood. Stability for mRNA was demonstrated for up to 2-year-old stains. As additional markers discovered in the future could be added to the analysis without any change to the basic protocol. Additional and more sensitive markers would contribute to the more reliable identification of a stain. Body fluids were collected from healthy volunteers. Detailed descriptions of materials and methods address sampling procedures; RNA extraction and reverse transcription; endpoint PCR; capillary electrophoresis; realtime PCR; DNA extraction, amplification, and detection; and enzymatic/immunologic tests. 3 tables, 4 figures, and 30 references