Messenger RNA profiling: a prototype method to supplant conventional methods for body fluid identification.
Conventional methods of body fluid identification use a variety of labor-intensive, technologically diverse techniques that are performed in a series, not parallel, manner and are costly in terms of time and sample. Theoretically, the identification of a body fluid may be made by determining a sufficient number of mRNAs that are expressed exclusively in cells that collectively comprise that body fluid. Advantages of an mRNA-based approach, compared to conventional biochemical methods of analysis, include greater specificity, simultaneous and semi-automatic analysis through a common assay format, improved timeliness, decreased sample consumption and compatibility with DNA extraction methodologies. In this report, we demonstrate that RNA is stable in biological stains and can be recovered in sufficient quantity and quality for analysis. Messenger RNA from the housekeeping genes S15, beta-actin and GAPDH was detected in blood, semen and saliva stains using a sensitive reverse transcriptase-polymerase chain reaction assay (RT-PCR). Additionally, we have identified a number of candidate tissue-specific genes, statherin, histatin 3, PRB1, PRB2 and PRB3 that may be useful for the positive identification of saliva. Messenger RNAs from these genes were detectable in saliva stains but not in blood or semen stains. Collectively these findings constitute the basis of a prototype RNA based assay system that may eventually supplant conventional methods for body fluid identification.
Juusola
Jane
J
Graduate Program in Biomolecular Science, University of Central Florida, PO Box 162366, Orlando, FL 32816-2366, USA.
Ballantyne
Jack
J
D000199:Actins / Q000235:genetics; D001774:Blood Chemical Analysis; D001801:Blood Stains; D004587:Electrophoresis, Agar Gel; D005554:Forensic Medicine / Q000379:methods*; D005656:Fungal Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D054808:Histatins; D006801:Humans; D008297:Male; D011506:Proteins / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis* / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics; D012463:Saliva / Q000737:chemistry*; D012471:Salivary Proteins and Peptides / Q000235:genetics; D012661:Semen / Q000737:chemistry*; D012697:Serine Endopeptidases / Q000235:genetics
D000199:Actins; D005656:Fungal Proteins; C068075:HTN3 protein, human; D054808:Histatins; D011506:Proteins; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D012471:Salivary Proteins and Peptides; C036004:ribosomal protein S15; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D012697:Serine Endopeptidases